片仔癀对人骨肉瘤阿霉素耐药细胞的作用及机制探讨
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摘要
目的
本实验以人骨肉瘤U20S细胞经ADM诱导建立的骨肉瘤ADM耐药细胞和相应的耐药细胞裸鼠移植瘤模型为研究对象,观察片仔癀对人骨肉瘤ADM耐药细胞的作用并探讨其可能的机制,为临床应用片仔癀逆转骨肉瘤耐药提供实验依据。
方法
1、人骨肉瘤U20S细胞ADM耐药细胞株的建立及其耐药机制
采用大剂量间隙作用、浓度梯度递增间隙作用建立人骨肉瘤U20S细胞ADM耐药株分别命名为U2OS/ADM1、U2OS/ADM2,光镜及透射电镜下观察细胞形态变化；MTT法绘制生长曲线计算倍增时间,测定其耐药指数及药物敏感性；FCM检测细胞内Rh123蓄积情况；RT-PCR检测MDR1、MRP1及Survivin mRNA的表达；Western blot检测P-gp、MRP1及Survivin蛋白的表达。
2、片仔癀对人骨肉瘤U2OS/ADM2细胞的作用
采用MTT法确定无细胞毒作用的片仔癀IC10浓度为联用浓度,MTT检测片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞增殖的影响,利用金氏公式求Q值,进行合用效果判断；Hoechst33258染色、Annexin V-FITC/PI双染FCM检测片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞凋亡的影响。
3、片仔癀逆转人骨肉瘤U2OS/ADM2细胞耐药的机制
采用FCM检测片仔癀对U2OS/ADM2细胞内Rh123蓄积的影响、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞内ADM蓄积的影响；RT-PCR检测不同浓度片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞Bcl-2、Bax. Survivin、MDR1mRNA表达的影响；Western blot去检测Bcl-2-、Bax、Survivin、P-gp蛋白的表达。
4、片仔癀对人骨肉瘤U2OS/ADM2细胞裸鼠移植瘤的作用
采用人骨肉瘤耐药株U2OS/ADM2细胞构建裸鼠皮下移植瘤模型,观察片仔癀、ADM、片仔癀与ADM联用对U2OS/ADM2细胞裸鼠移植瘤体积、重量的影响,绘制肿瘤生长曲线,计算肿瘤抑制率(IR),TUNEL法检测移植瘤细胞凋亡情况,RT-PCR检测Bcl-2、Bax、MDR1mRNA在移植瘤中的表达。
结果
1、成功建立了人骨肉瘤U20S细胞ADM耐药株U2OS/ADM1、U2OS/ADM2,镜下观察U2OS/ADM1、U2OS/ADM2细胞较亲本细胞U2OS在形态上发生了显著变化,透射电镜观察发现耐药株细胞内空泡增多,核糖体增多,粗面内质网扩张；U2OS/ADM1与U2OS/ADM2细胞倍增时间延长,与U2OS细胞相比差异明显(PF)<0.01)；U2OS/ADMK、U2OS/ADM2细胞的耐药指数(RI)分别为79.63和91.06,同时对紫杉醇、顺铂交叉耐药；细胞内Rh123蓄积量明显低于U2OS细胞(P<0.01),而两种耐药株对Rh123的蓄积无明显差异(P>0.05)；U2OS/ADM1、U2OS/ADM2细胞中MDR1/P-gp、MRP1及Survivin基因及蛋白的表达均明显高于U2OS细胞(P<0.05),且U2OS/ADM2细胞中MRP1的表达较U2OS/ADM1细胞明显上调(P<0.05)。
2、片仔癀对U2OS/ADM2细胞增殖具有抑制作用,呈时间和剂量依赖性；片仔癀可促进U2OS/ADM2细胞凋亡,凋亡率随片仔癀浓度的增大而增加；U2OS细胞和U2OS/ADM2细胞对片仔癀的IC50无明显差异(P>0.05),U2OS/ADM2细胞对片仔癀无交叉耐药性。片仔癀对U2OS/ADM2细胞无毒性作用的药物浓度(IC10)为0.4mg/ml。 U2OS/ADM2细胞对ADM的IC50为(14.57±0.15)μg/ml,0.4mg/ml片仔癀与ADM联用后其对ADM的IC50为(6.75±0.29)μ g/ml,逆转指数为2.16；Q值为2.29;0.4mg/ml片仔癀与ADM联用后细胞凋亡率与单用ADM相比明显增加(P<0.05)。
3、片仔癀可增加细胞内P-gp底物Rh123的蓄积,下调U2OS/ADM2细胞中Bcl-2、Survivin、MDR1/P-gp的表达,上调Bax表达,且呈剂量依赖性,与对照组相比有明显差异(P<0.05)；0.4mg/ml片仔癀与ADM联用后细胞内ADM荧光强度为对照组的1.27倍(P<0.05),与对照组、单用ADM组相比,联用组Bcl-2、Survivin、MDR1/P-gp的表达下调,而Bax的表达上调,差异具有统计学意义(P<0.05)。
4、成功建立了U2OS/ADN2细胞裸鼠移植瘤模型,片仔癀与ADM联用组裸鼠移植瘤的体积、重量与片仔癀组、ADM组相比明显降低(P<0.05)。TUNEL检测显示联用组凋亡指数明显高于片仔癀组、ADM组,差异有统计学意义(P<0.01)。RT-PCR结果显示片仔癀与ADM联用可明显上调Bax mRNA的表达,降低Bcl-2、MDR1mRNA的表达,与单用片仔癀和单用ADM相比差异具有统计学意义(P<0.05)。
结论
1、采用ADM大剂量间隙作用、浓度梯度递增间隙作用法均可建立人骨肉瘤ADM耐药细胞株,耐药机制与MDR1、MRP1、Survivin过表达有关；不同建立方式存在一定差异,浓度梯度递增间隙作用法建立的耐药细胞耐药性更明显。
2、片仔癀能够有效抑制U2OS/ADM2细胞的增殖,促进其凋亡,呈时间和剂量依赖性；无细胞毒作用的片仔癀与ADM可产生协同作用,提高U2OS/ADM2细胞对ADM的敏感性,表明片仔癀具有逆转U2OS/ADM2细胞耐药的作用。其机制与抑制P-gp的表达,改变Bcl-2与Bax的比例,下调Survivin的表达有关。
3、片仔癀与ADM联用可明显抑制U2OS/ADM2细胞裸鼠移植瘤的生长,促进其凋亡,其机制与上调Bax表达,下调Bcl-2、MDR1的表达有关。
Objective
Using human osteosarcoma adriamycin resistant U20S/ADM cells and the corresponding animal model for the study, to evaluate the effects of Pien Tze Huang on U2OS/ADM2cells and the associated mechanisms, to provide experimental basis for Pien Tze Huang to reverse osteosarcoma drug-resistance in clinical.
Methods
1. Establishment of an adriamycin resistant human osteosarcoma U2OS/ADM cells and explore its drug resistance mechanism.
Two resistant cell lines U20S/ADM1and U2OS/ADM2were established by impacting with high dose of adriamycin for short time and gradually increasing concentration of adriamycin from the parent cell line U2OS in vitro, observed their morphological features by optical microscope and transmission electron microscope, checked their resistance index (RI) and drug sensitivity by MTT assay, gathered their growth curve and calculated their doubling time. The accumulation of Rh123was evaluated by flow cytometry. Expressions of MDR1/P-gp、MRP1and Survivin were measured quantitatively by RT-PCR and Western blot.
2. Study of reversal effects of Pien Tze Huang (PZH) on U2OS/ADM2cells
PZH alone or combined with adriamycin were used to treated U2OS/ADM2cells, the growth inhibitions were detected by MTT assays, the combination effect of PZH with ADM on U2OS/ADM2cells was calculated by index Q and reversal index; the apoptosis was detected by Hoechst33258staining assay and Annexin V-FITC/PI double staining analysis by flow cytometry.
3. Study of the reversal mechanisms of PZH on U2OS/ADM2cells
PZH alone or combined with adriamycin were used to treated U2OS/ADM2cells, the accumulation of Rh123or ADM was evaluated by flow cytometry. Expressions of Bcl-2、 Bax、Survivin and MDRl/P-gp were studied by RT-PCR and Western blot.
4. Study on the reversal effects of PZH on U2OS/ADM xenograft in nude mice
The nude mice modes were built by injecting U2OS/ADM2cells in nude mice subcutaneous. PZH or ADM or PZH combined with adriamycin were used to treated the nude mice. Tumors volume and tumors weight were measured during the drug therapy, the growth curve was gathered and tumor volume and weight inhibition rate were calculated. The apoptosis was detected by In situ end labeling (TUNEL) and the expressions of Bcl-2、Bax、 MDR1mRNA were measured by RT-PCR.
Result
1. The RI of U2OS/ADM1and U2OS/ADM2cells were79.63and91.06to adriamycin. They also demonstrated cross resistance to paclitaxel and cisplatin. Both resistant cell lines grew slowly, their doubling time were prolonged (P<0.01,vs U2OS cells) and exhibited changes in morphology. The accumulation of Rh123was lower in two resistant cell lines than in the parent cells (P<0.01, vs U2OS cells). The expressions of MDR1, MRP1and Survivin were enhanced in both resistant cell lines (P<0.05, vs U2OS cells). The expression of MRP1was higher in U2OS/ADM2cells than in U2OS/ADM1cells (P<0.05).
2. The IC50value of PZH has no significant difference on the U2OS cells and U2OS/ADM2cells, indicating that drug resistant U2OS/ADM2cells have no cross-resistance to PZH. The proliferation of U2OS/ADM2cells was inhibited by PZH in a dose-time-dependent manner. The IC10of PZH to U2OS/ADM2cells is0.4mg/ml. The U2OS/ADM cells growth inhibition rate was significantly increased compared the combined effect of0.4mg/ml PZH and ADM with PZH and ADM separately. The IC50value of ADM in U2OS/ADM cells was (14.57±0.15)μg/ml, it decreased to (6.75±0.29)μg/ml after combined with PZH, the reverse index is2.16, the combined index Q was2.29>1.15. The percentage of U2OS/ADM2cells apoptosis were significantly increased by PZH in a dose-dependent manner. Cooperated the apoptosis rate of PZH and ADM with PZH and ADM separately, the combination of PZH and ADM significantly increased (P<0.05)
3. PZH attenuated the expressions of Bcl-2, Survivin and MDR1/P-gp, activated the expression of Bax at mRNA and protein levels, enhanced the accumulation of Rh123in U2OS/ADM2cells in a does-dependent manner (P<0.05, vs control). The fluorescence intensity of ADM in U2OS/ADM2cells treated by0.4mg/ml PZH combined with ADM was increased1.27times compared by AMD. The mRNA and protein expressions of Bcl-2、 Survivin and MDRl/P-gp were downregulated and the mRNA and protein expression of Bax was upregulated in0.4mg/ml PZH combined with ADM group (P<0.05, vs. control)
4. The tumor models of human osteosarcoma U2OS/ADM2cells xenograft were successfully constructed. Compared the combined effect of PZH and ADM group with PZH and ADM group separately, volume and weight of U2OS/ADM2cells xenograft were significantly decreased (P<0.05), the apoptotic index was increased detected by TUNEL assay (P<0.01). RT-PCR results showed that the mRNA expression level of Bcl-2and MDR1were downregulated and the mRNA expression level of Bax was upregulated in the PZH and ADM group, compared to the PZH group and ADM group (P<0.05)
Conclusion
1. Using high dose of adriamycin for short time and gradually increasing concentration of adriamycin from the parent cell line U2OS in vitro can establishment the adriamycin resistant human osteosarcoma cell lines U2OS/ADM1and U2OS/ADM2, the drug-resistance mechanisms maybe relate to overexpressions of MDR1/P-gp, MRP1and Survivin. The resistant ability of U2OS/ADM2cells was higher than U2OS/ADM1cells.
2. PZH can inhibit the growth of U2OS/ADM2cells and promote apoptosis in time and dose-dependent manner, and PZH has a synergistic effect with ADM, increase the sensitivity of U2OS/ADM2cells to ADM. The mechanism is related to inhibit the expression of P-gp, change the proportion of Bcl-2and Bax, and down regulate the expression of Survivin.
3. PZH can enhance the anti-tumor effect of ADM on U2OS/ADM2cells xenograft, the mechanism is related to downregulate the expressions of Bcl-2and MDR1gene and upregulate the expression of Bax gene.
本实验以人骨肉瘤U20S细胞经ADM诱导建立的骨肉瘤ADM耐药细胞和相应的耐药细胞裸鼠移植瘤模型为研究对象,观察片仔癀对人骨肉瘤ADM耐药细胞的作用并探讨其可能的机制,为临床应用片仔癀逆转骨肉瘤耐药提供实验依据。
方法
1、人骨肉瘤U20S细胞ADM耐药细胞株的建立及其耐药机制
采用大剂量间隙作用、浓度梯度递增间隙作用建立人骨肉瘤U20S细胞ADM耐药株分别命名为U2OS/ADM1、U2OS/ADM2,光镜及透射电镜下观察细胞形态变化；MTT法绘制生长曲线计算倍增时间,测定其耐药指数及药物敏感性；FCM检测细胞内Rh123蓄积情况；RT-PCR检测MDR1、MRP1及Survivin mRNA的表达；Western blot检测P-gp、MRP1及Survivin蛋白的表达。
2、片仔癀对人骨肉瘤U2OS/ADM2细胞的作用
采用MTT法确定无细胞毒作用的片仔癀IC10浓度为联用浓度,MTT检测片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞增殖的影响,利用金氏公式求Q值,进行合用效果判断；Hoechst33258染色、Annexin V-FITC/PI双染FCM检测片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞凋亡的影响。
3、片仔癀逆转人骨肉瘤U2OS/ADM2细胞耐药的机制
采用FCM检测片仔癀对U2OS/ADM2细胞内Rh123蓄积的影响、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞内ADM蓄积的影响；RT-PCR检测不同浓度片仔癀、无细胞毒作用浓度的片仔癀与ADM联用对U2OS/ADM2细胞Bcl-2、Bax. Survivin、MDR1mRNA表达的影响；Western blot去检测Bcl-2-、Bax、Survivin、P-gp蛋白的表达。
4、片仔癀对人骨肉瘤U2OS/ADM2细胞裸鼠移植瘤的作用
采用人骨肉瘤耐药株U2OS/ADM2细胞构建裸鼠皮下移植瘤模型,观察片仔癀、ADM、片仔癀与ADM联用对U2OS/ADM2细胞裸鼠移植瘤体积、重量的影响,绘制肿瘤生长曲线,计算肿瘤抑制率(IR),TUNEL法检测移植瘤细胞凋亡情况,RT-PCR检测Bcl-2、Bax、MDR1mRNA在移植瘤中的表达。
结果
1、成功建立了人骨肉瘤U20S细胞ADM耐药株U2OS/ADM1、U2OS/ADM2,镜下观察U2OS/ADM1、U2OS/ADM2细胞较亲本细胞U2OS在形态上发生了显著变化,透射电镜观察发现耐药株细胞内空泡增多,核糖体增多,粗面内质网扩张；U2OS/ADM1与U2OS/ADM2细胞倍增时间延长,与U2OS细胞相比差异明显(PF)<0.01)；U2OS/ADMK、U2OS/ADM2细胞的耐药指数(RI)分别为79.63和91.06,同时对紫杉醇、顺铂交叉耐药；细胞内Rh123蓄积量明显低于U2OS细胞(P<0.01),而两种耐药株对Rh123的蓄积无明显差异(P>0.05)；U2OS/ADM1、U2OS/ADM2细胞中MDR1/P-gp、MRP1及Survivin基因及蛋白的表达均明显高于U2OS细胞(P<0.05),且U2OS/ADM2细胞中MRP1的表达较U2OS/ADM1细胞明显上调(P<0.05)。
2、片仔癀对U2OS/ADM2细胞增殖具有抑制作用,呈时间和剂量依赖性；片仔癀可促进U2OS/ADM2细胞凋亡,凋亡率随片仔癀浓度的增大而增加；U2OS细胞和U2OS/ADM2细胞对片仔癀的IC50无明显差异(P>0.05),U2OS/ADM2细胞对片仔癀无交叉耐药性。片仔癀对U2OS/ADM2细胞无毒性作用的药物浓度(IC10)为0.4mg/ml。 U2OS/ADM2细胞对ADM的IC50为(14.57±0.15)μg/ml,0.4mg/ml片仔癀与ADM联用后其对ADM的IC50为(6.75±0.29)μ g/ml,逆转指数为2.16；Q值为2.29;0.4mg/ml片仔癀与ADM联用后细胞凋亡率与单用ADM相比明显增加(P<0.05)。
3、片仔癀可增加细胞内P-gp底物Rh123的蓄积,下调U2OS/ADM2细胞中Bcl-2、Survivin、MDR1/P-gp的表达,上调Bax表达,且呈剂量依赖性,与对照组相比有明显差异(P<0.05)；0.4mg/ml片仔癀与ADM联用后细胞内ADM荧光强度为对照组的1.27倍(P<0.05),与对照组、单用ADM组相比,联用组Bcl-2、Survivin、MDR1/P-gp的表达下调,而Bax的表达上调,差异具有统计学意义(P<0.05)。
4、成功建立了U2OS/ADN2细胞裸鼠移植瘤模型,片仔癀与ADM联用组裸鼠移植瘤的体积、重量与片仔癀组、ADM组相比明显降低(P<0.05)。TUNEL检测显示联用组凋亡指数明显高于片仔癀组、ADM组,差异有统计学意义(P<0.01)。RT-PCR结果显示片仔癀与ADM联用可明显上调Bax mRNA的表达,降低Bcl-2、MDR1mRNA的表达,与单用片仔癀和单用ADM相比差异具有统计学意义(P<0.05)。
结论
1、采用ADM大剂量间隙作用、浓度梯度递增间隙作用法均可建立人骨肉瘤ADM耐药细胞株,耐药机制与MDR1、MRP1、Survivin过表达有关；不同建立方式存在一定差异,浓度梯度递增间隙作用法建立的耐药细胞耐药性更明显。
2、片仔癀能够有效抑制U2OS/ADM2细胞的增殖,促进其凋亡,呈时间和剂量依赖性；无细胞毒作用的片仔癀与ADM可产生协同作用,提高U2OS/ADM2细胞对ADM的敏感性,表明片仔癀具有逆转U2OS/ADM2细胞耐药的作用。其机制与抑制P-gp的表达,改变Bcl-2与Bax的比例,下调Survivin的表达有关。
3、片仔癀与ADM联用可明显抑制U2OS/ADM2细胞裸鼠移植瘤的生长,促进其凋亡,其机制与上调Bax表达,下调Bcl-2、MDR1的表达有关。
Objective
Using human osteosarcoma adriamycin resistant U20S/ADM cells and the corresponding animal model for the study, to evaluate the effects of Pien Tze Huang on U2OS/ADM2cells and the associated mechanisms, to provide experimental basis for Pien Tze Huang to reverse osteosarcoma drug-resistance in clinical.
Methods
1. Establishment of an adriamycin resistant human osteosarcoma U2OS/ADM cells and explore its drug resistance mechanism.
Two resistant cell lines U20S/ADM1and U2OS/ADM2were established by impacting with high dose of adriamycin for short time and gradually increasing concentration of adriamycin from the parent cell line U2OS in vitro, observed their morphological features by optical microscope and transmission electron microscope, checked their resistance index (RI) and drug sensitivity by MTT assay, gathered their growth curve and calculated their doubling time. The accumulation of Rh123was evaluated by flow cytometry. Expressions of MDR1/P-gp、MRP1and Survivin were measured quantitatively by RT-PCR and Western blot.
2. Study of reversal effects of Pien Tze Huang (PZH) on U2OS/ADM2cells
PZH alone or combined with adriamycin were used to treated U2OS/ADM2cells, the growth inhibitions were detected by MTT assays, the combination effect of PZH with ADM on U2OS/ADM2cells was calculated by index Q and reversal index; the apoptosis was detected by Hoechst33258staining assay and Annexin V-FITC/PI double staining analysis by flow cytometry.
3. Study of the reversal mechanisms of PZH on U2OS/ADM2cells
PZH alone or combined with adriamycin were used to treated U2OS/ADM2cells, the accumulation of Rh123or ADM was evaluated by flow cytometry. Expressions of Bcl-2、 Bax、Survivin and MDRl/P-gp were studied by RT-PCR and Western blot.
4. Study on the reversal effects of PZH on U2OS/ADM xenograft in nude mice
The nude mice modes were built by injecting U2OS/ADM2cells in nude mice subcutaneous. PZH or ADM or PZH combined with adriamycin were used to treated the nude mice. Tumors volume and tumors weight were measured during the drug therapy, the growth curve was gathered and tumor volume and weight inhibition rate were calculated. The apoptosis was detected by In situ end labeling (TUNEL) and the expressions of Bcl-2、Bax、 MDR1mRNA were measured by RT-PCR.
Result
1. The RI of U2OS/ADM1and U2OS/ADM2cells were79.63and91.06to adriamycin. They also demonstrated cross resistance to paclitaxel and cisplatin. Both resistant cell lines grew slowly, their doubling time were prolonged (P<0.01,vs U2OS cells) and exhibited changes in morphology. The accumulation of Rh123was lower in two resistant cell lines than in the parent cells (P<0.01, vs U2OS cells). The expressions of MDR1, MRP1and Survivin were enhanced in both resistant cell lines (P<0.05, vs U2OS cells). The expression of MRP1was higher in U2OS/ADM2cells than in U2OS/ADM1cells (P<0.05).
2. The IC50value of PZH has no significant difference on the U2OS cells and U2OS/ADM2cells, indicating that drug resistant U2OS/ADM2cells have no cross-resistance to PZH. The proliferation of U2OS/ADM2cells was inhibited by PZH in a dose-time-dependent manner. The IC10of PZH to U2OS/ADM2cells is0.4mg/ml. The U2OS/ADM cells growth inhibition rate was significantly increased compared the combined effect of0.4mg/ml PZH and ADM with PZH and ADM separately. The IC50value of ADM in U2OS/ADM cells was (14.57±0.15)μg/ml, it decreased to (6.75±0.29)μg/ml after combined with PZH, the reverse index is2.16, the combined index Q was2.29>1.15. The percentage of U2OS/ADM2cells apoptosis were significantly increased by PZH in a dose-dependent manner. Cooperated the apoptosis rate of PZH and ADM with PZH and ADM separately, the combination of PZH and ADM significantly increased (P<0.05)
3. PZH attenuated the expressions of Bcl-2, Survivin and MDR1/P-gp, activated the expression of Bax at mRNA and protein levels, enhanced the accumulation of Rh123in U2OS/ADM2cells in a does-dependent manner (P<0.05, vs control). The fluorescence intensity of ADM in U2OS/ADM2cells treated by0.4mg/ml PZH combined with ADM was increased1.27times compared by AMD. The mRNA and protein expressions of Bcl-2、 Survivin and MDRl/P-gp were downregulated and the mRNA and protein expression of Bax was upregulated in0.4mg/ml PZH combined with ADM group (P<0.05, vs. control)
4. The tumor models of human osteosarcoma U2OS/ADM2cells xenograft were successfully constructed. Compared the combined effect of PZH and ADM group with PZH and ADM group separately, volume and weight of U2OS/ADM2cells xenograft were significantly decreased (P<0.05), the apoptotic index was increased detected by TUNEL assay (P<0.01). RT-PCR results showed that the mRNA expression level of Bcl-2and MDR1were downregulated and the mRNA expression level of Bax was upregulated in the PZH and ADM group, compared to the PZH group and ADM group (P<0.05)
Conclusion
1. Using high dose of adriamycin for short time and gradually increasing concentration of adriamycin from the parent cell line U2OS in vitro can establishment the adriamycin resistant human osteosarcoma cell lines U2OS/ADM1and U2OS/ADM2, the drug-resistance mechanisms maybe relate to overexpressions of MDR1/P-gp, MRP1and Survivin. The resistant ability of U2OS/ADM2cells was higher than U2OS/ADM1cells.
2. PZH can inhibit the growth of U2OS/ADM2cells and promote apoptosis in time and dose-dependent manner, and PZH has a synergistic effect with ADM, increase the sensitivity of U2OS/ADM2cells to ADM. The mechanism is related to inhibit the expression of P-gp, change the proportion of Bcl-2and Bax, and down regulate the expression of Survivin.
3. PZH can enhance the anti-tumor effect of ADM on U2OS/ADM2cells xenograft, the mechanism is related to downregulate the expressions of Bcl-2and MDR1gene and upregulate the expression of Bax gene.
引文
[1]Ottaviani G and Jaffe N. The epidemiology of osteosarcoma [J]. Cancer Treat Res,2009,152:3-13.
[2]Mirabello L, Troisi RJ and Savage SA. Osteosarcoma incidence and survival rates from 1973 to 2004: data from the Surveillance, Epidemiology, and End Results Program [J]. Cancer,2009,115 (7):1531-1543.
[3]Rajani R and Gibbs CP. Treatment of bone tumors [J]. Surg Pathol Clin,2012,5 (1):301-318.
[4]Ando K, Heymann MF, Stresing V, et al. Current therapeutic strategies and novel approaches in osteosarcoma [J].Cancers,2013,5(2):591-616.
[5]Chou AJ, Gorlick R. Chemotherapy resistance in osteosarcoma:current challenges and future directions [J]. Expert Rev Anticancer Ther,2006,6(7):1075-1085.
[6]Schwartz CL, Gorlick R, Teot L, et al. Children's Oncology Group:Multiple drug resistance in osteogenic sarcoma:INTO133 from the Children's Oncology Group [J]. J Clin Oncol,2007,25(15): 2057-2062.
[7]Baguley BC. Multiple drug resistance mechanisms in cancer [J]. Mol Biotechnol,2010,46(3):308-316.
[8]Liu FS. Mechanisms of chemotherapeutic drug resistance in cancer therapy--a quick review [J]. Taiwan J Obstet Gynecol,2009,48(3):239-44.
[9]Shukla S, Ohnuma S, Ambudkar SV. Improving cancer chemotherapy with modulators of ABC drug transporters [J]. Curr Drug Targets,2011,12(5):621-630.
[10]杨华,黄文荣,于力.新型抗肿瘤药物的临床应用及研究进展[J].中国新药杂志,2010,19(6)：480-485.
[11]Li-Weber M. Targeting apoptosis pathways in cancer by Chinese medicine [J]. Cancer Lett,2013, 332(2):304-312.
[12]Yang L, Wei DD, Chen Z, et al. Reversal effects of traditional Chinese herbs on multidrug resistance in cancer cells [J]. Nat Prod Res,2011,25(19):1885-1889.
[13]王辉,孙桂芝.孙桂芝治疗骨肉瘤经验[J].世界中医药,2012,7(1)：21-22.
[14]孟丽萍,顾雪峰.片仔癀联合治疗毒热瘀结型原发性肝癌免疫指标变化的临床观察[J].九江医学,2008,23(1)：31-37.
[15]林明和,朱德增.片仔癀联合化疗治疗热毒瘀结型晚期结肠癌临床研究[J].福建中医药,2102,43(1)：8-9.
[16]Lu L, Wai MS, Yew DT, et al. Pien Tze Huang, a composite Chinese traditional herbal extract, affects survival of neuroblastoma cells [J]. Int J Neurosci,2009,119(2):255-262.
[17]Lin Jiu-mao, Wei Li-hui, Chen You-qin, et al. Pien Tze Huang-induced apoptosis in human colon cancer HT-29 cells is associated with regulation the Bcl-2 family and activation of caspase3 [J].Chin J Integr Med,2011, 11(9):685-690.
[18]Shen A, Hong F, Liu L, et al. Pien Tze Huang inhibits the proliferation of human colon carcinoma cells by arresting Gl/S cell cycle progression[J]. Oncol Lett.2012,4(4):767-770.
[19]Shen A, Chen Y, Hong F, et al, Pien Tze Huang suppresses IL-6-inducible STAT3 activation in human colon carcinoma cells through induction of SOCS3 [J]. Oncol Rep.2012,28(6):2125-30.
[20]张俐,曾启清,林建华.P27基因联合中药片仔癀对人骨肉瘤抑制作用观察[J].中华中医药杂志,2009,24(4)：511-515.
[21]张俐,于波,林建华.中药片仔癀胶囊对骨肉瘤细胞诱导凋亡的作用[J].中国骨伤,2009, 22(4):265-268.
[22]张燕,王琪鸿,张俐,等.片仔癀对人骨肉瘤细胞U20S细胞周期的影响[J].江西中医学院学报,2012,24(5)：61-63.
[23]杨德建.片仔癀对裸鼠原位移植骨肉瘤的作用及机制研究[D],福建：福建中医药大学,2013.
[24]Wang C, Guo LB, Ma JY, et al. Establishment and characterization of a paclitaxel-resistant human esophageal carcinoma cell line [J]. Int J Oncol,2013,43(5):1607-1617.
[25]Zhang X, Yashiro M, Qiu H, et al. Establishment and characterization of multidrug-resistant gastric cancer cell lines [J]. Anticancer Res,2010,30(3):915-921.
[26]赵斌,葛金芳,朱娟娟,等小议在MTT法测细胞增殖抑制率中IC50的计算方法[J].安徽医药,2007,11(9)：834-835.
[27]Ando K, Heymann MF, Stresing V, et al. Current therapeutic strategies and novel approaches in osteosarcoma [J]. Cancers (Basel),2013,5(2):591-616.
[28]Scheetze SM. Chemotherapy in the management of osteosarcoma and E wing's sarcoma [J]. J Natl Compr Canc Netw,2007,5(4):449-455.
[29]Saraswathy M, Gong S. Different strategies to overcome multidrug resistance in cancer [J]. Biotechnol Adv,2013,31(8):1397-1407.
[30]Niu BH, Wang JJ, Xi Y, et al. The establishment and characterization of adriamycin-resistant cell lines derived from Saos-2 [J]. Medical Science Monitor,2010,16(6):BR184-192.
[31]Uchibori K, Kasamatsu A, Sunaga M, et al. Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines [J]. Int J Oncol,2012,40(4):1005-1010.
[32]Ahrens AK, Chaturvedi NK, Nordgren TM, et al. Establishment and characterization of therapy-resistant mantle cell lymphoma cell lines derived from different tissue sites [J]. Leuk Lymphoma, 2012,53(11):2269-2278.
[33]Carrle D, Bielack SS. Current strategies of chemotherapy in osteosarcoma [J]. Int Orthop,2006, 30(6):445-451.
[34]Rebbaa A. Targeting senescence pathways to reverse drug resistance in cancer [J]. Cancer Lett,2005, 219(1):1-13.
[35]Saretzki G.Cellular senescence in the development and treatment of cancer [J]. Curr Pharm Des,2010, 16(1):79-100.
[36]Juliao RL and Linvg V. A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants [J]. Biochim Biophys Acta,1976,455:152-162.
[37]Binkhathlan Z and Lavasanifar A. P-glycoprotein inhibition as a therapeutic approach for overcoming multidrug resistance in cancer:current status and future perspectives [J]. Curr Cancer Drug Targets,2013, 13(3):326-346.
[38]Mueller CF, Wassmann K, Widder JD, et al. Multidrug resistance protein-1 affects oxidative stress, endothelial dysfunction, and atherogenesis via leukotriene C4 export [J]. Circulation,2008, 117(22):2912-2918.
[39]Johnstone RW, Ruefli AA, Tainton KM, et al. A role for P-glycoprotein in regulating cell death [J]. Leuk Lymphoma,2000,38(1-2):1-11.
[40]Yamagishi T, Sahni S, Sharp DM, et al. P-glycoprotein mediates drug resistance via a novel mechanism involving lysosomal sequestration [J]. J Biol Chem,2013,288(44):31761-31771.
[41]Gomes CM, van Paassen H, Romeo S, et al. Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA analysis and functional radiotracer studies [J]. Nuclear Medicine and Biology,2006,33(7):831-840.
[42]Cole SP, Bhardwai G, Gerlach JH, et al. Over-expression of a transporters gene in a multidrug-resistant human lung cancer cell line [J]. Science,1992,5088:1650-1654.
[43]Leslie EM, Deeley RG, Cole SP. Multidrug resistance proteins:role of P-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense [J]. Toxicol Appl Pharmacol,2005,204(3):216-237.
[44]Keppler D. Multidrug resistance proteins (MRPs, ABCCs):importance for pathophysiology and drug therapy [J]. Handb Exp Pharmacol,2011,201:299-323.
[45]Homolya L, Varadi A and Sarkadi B. Multidrug resistance-associated proteins:Export pumps for conjugates with glutathione, glucuronate or sulfate [J]. Biofactors,2003,17(1-4):103-114.
[46]Wang L, Zhang GM, Feng Zh. Down-regulation of survivin expression reversed multidrug resistance in adriamycin-resistant HL-60/ADR cell line[J]. Acta Pharmacol Sin,2003,24(12):1235-1240.
[47]Ambrosini G, Adida C, Altieri DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med,1997,3(8):917-921.
[48]Shoeneman JK, Ehrhart EJ 3rd, Eickhoff JC, et al. Expression and function of survivin in canine osteosarcoma [J]. Cancer Res,2012,72(1):249-259.
[49]Zou J, Gan M, Mao N, et al. Sensitization of osteosarcoma cell line SaOS-2 to chemotherapy by downregulating survivin [J]. Arch Med Res,2010,41(3):162-9.
[50]黄芳.植物多酚类化合物联合阿霉素抑制MCF-7/ADM细胞的筛选及白藜芦醇逆转耐药性作用研究[D],福建：福建医科大学,2009：34-35.
[51]Chou AJ, Geller DS, Gorlick R. Therapy for osteosarcoma:where do we go from here? [J]. Paediatr Drugs,2008,10(5):315-327.
[52]Ayan I, Kebudi R, Ozger H. Childhood osteosarcoma:multimodal therapy in a single-institution Turkish series [J]. Cancer Treat Res.2009,152:319-338.
[53]牛晓辉,徐海荣.骨肉瘤的化疗进展[J].中国癌症杂志,2012,22(9)：663-669.
[54]凌昌全.恶性肿瘤病因病机之我见[J].中医杂志,2009,50(10)：952-953.
[55]吴薏婷,吴锦燕.陈庆强主任治疗恶性肿瘤经验[J].辽宁中医药大学学报,2011,13(5)：164-165.
[56]潘万刚.中药三棱祁甲汤治疗骨肉瘤22例临床疗效分析[J].中华临床医学杂志,2005,6(9)：88-90.
[57]邹存华,付婷婷,鲁强,等.三七总皂苷抑制Hela细胞增殖的实验研究[J].中华肿瘤防治杂志2012,19(12)：888-891.
[58]杨茹萍,陈彤,陈亚娟,等.三七提取物的体外抗肿瘤药理作用及其成分分析[J].昆明医学院学报,2011,9：4-6.
[59]Wang CZ, Luo XJ, Zhang B, et al. Notoginseng enhances anticancer effect of 5-fluotouiacill on human colorectal cancer cells[J]. Cancer Chemother Pharmacol,2007,60(1):69-79.
[60]黄芳.牛黄对胃癌细胞BCG-823增殖抑制、诱导凋亡的实验研究[D].湖南：中南大学,2013：29.
[61]吴志远,李芳芳,张昆,等.人工牛黄对小鼠乳腺癌肺转移的影响[J].中草药,2012,43(10)：2013-2016.
[62]多明荷,孙莉娜,应森林,等.西黄丸通过ERK/MAPK信号通路对人结肠癌裸鼠移植瘤的影响[J].中华中医药杂志,2013,28(10)：3055-3058.
[63]Massart C, Gibassier J, Raoul M, et al. Cyclosporin A, verapamil and S9788 reverse doxorubicin resistance in a human medullary thyroid carcinoma cell line [J]. Anticancer Drugs,1995, 6(1):135-46.
[64]O'Brien MM, Lacayo NJ, Lum BL,et al. Phase I study of valspodar (PSC-833) with mitoxantrone and etoposide in refractory and relapsed pediatric acute leukemia:a report from the Children's Oncology Group p [J]. Pediatr Blood Cancer,2010,54(5):694-702.
[65]Bottova I, Sauder U, Olivieri V, et al. The P-glycoprotein inhibitor GF120918 modulates Ca2+-dependent processes and lipid metabolism in Toxoplasma gondii [J]. PLoS One,2010,5(4):el0062.
[66]Callies S, de Alwis DP, Harris A, et al. A population pharmacokinetic model for paclitaxel in the presence of a novel P-gp modulator, Zosuquidar Trihydrochloride (LY335979) [J].Br J Clin Pharmacol,2003,56(1):46-56.
[67]Jia L, Ma S, Hou X, et al. The synergistic effects of traditional Chinese herbs and radiotherapy for cancer treatment [J]. Oncol Lett,2013,5(5):1439-1447.
[68]Wu G, Chu J, Huang Z, et al. Xiao Jin Wan, a traditional Chinese herbal formula, inhibits proliferation via arresting cell cycle progression at the G2/M phase and promoting apoptosis via activating the mitochondrial-dependent pathway in U-2OS human osteosarcoma cells [J]. Int J Oncol Mar,2013,42(3): 1070-1080.
[69]吕仙梅,郑坚,朱莹杰,等.中医药联合化疗对大肠癌、期患者生存期的影响[J].中国中西结合杂志,2012,32(9)：1166-1170.
[70]余文燕,王国娟,许建华,等.肠胃清颗粒协同卡铂对人结肠癌耐药细胞增殖及凋亡的影响[J].中医杂志,2013,54(21)：1854-1858.
[71]Deng S, Hu B, An HM, et al. Teng-Long-Bu-Zhong-Tang, a Chinese herbal formula, enhances anticancer effects of 5--Fluorouracil in CT26 colon carcinoma [J]. BMC Complement Altern Med,2013, 13:128.
[72]肖扬,王万春.姜黄素对人骨肉瘤细胞株/阿霉素多药耐药逆转作用的实验研究[J].卫生研究,2011,40(1)103-106.
[73]Yang L, Wei DD, Chen Z, et al. Reversal of multidrug resistance in human breast cancer cells by Curcuma wenyujin and Chrysanthemum indicum [J]. Phytomedicine,2011,18(8-9):710-718.
[74]Zhu X, Sui M, Fan W. In vitro and in vivo characterizations of tetrandrine on the reversal of P-glycoprotein-mediated drug resistance to paclitaxel [J]. Anticancer Res.2005,25(3B):1953-1962.
[75]王进,沈靖南.粉防己碱逆转骨肉瘤细胞系多药耐药性的研究[J].实用临床医学,2005,6(7)：7-10.
[76]Xie X, Yin J, Jia Q, Wang J, et al. Quercetin induces apoptosis in the methotrexate-resistant osteosarcoma cell line U2-OS/MTX300 via mitochondrial dysfunction and dephosphorylation of Akt [J]. Oncol Rep,2011,26(3):687-693.
[77]余文燕,孙珏,徐建华,等.肠胃清药物血清协同奥沙利铂对人结肠癌耐药细胞增殖及凋亡的影响[J].中国实验方剂学杂志,,2013,19(10)：182-186.
[78]聂昊,王晖.P糖蛋白在不同组织中的分布与功能研究进展[J],广东药学院学报,2012,28(4)：456-460.
[79]Leslie EM, Deeley RG, Cole SP, et al. Multidrug resistance proteins:role of P-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense [J], Toxicol Appl Pharmacol.2005,204(3):216-237.
[80]Gomes CM, van Paassen H, Romeo S, et al. Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA analysis and functional radiotracer studies [J]. Nucl Med Biol,2006,33(7): 831-840.
[81]Serra M, Scotlandi K, Reverter-Branchat G, et al. Value of P-Glycoprotein and Clinicopathologic Factors as the Basis for New Treatment Strategies in High-Grade Osteosarcoma of the Extremities [J]. J Clin Oncol,2003,21(3):536-542.
[82]Sauna ZE, Ambudkar SV. About a switch:how P-glycoprotein (ABCB1) harnesses the energy of ATP binding and hydrolysis to do mechanical work [J]. Mol Cancer Ther,2007,6(1):13-23.
[83]Rosenberg MF, Kamis AB, Callaghan R, et al. Three-dimensional structures of the mammalian multidrug resistance P-glycoprotein demonstrate major conformational changes in the transmembrane domains upon nucleotide binding [J]. J Biol Chem,2003,278(10):8294-8299.
[84]Johnstone RW, Ruefli AA, Tainton KM, et al. A role for P-glycoprotein in regulating cell death [J]. Leuk Lymphoma,2000,38(1-2):1-11.
[85]Cappellini A, Chiarini F, Ognibene A, et al. The cyclin-dependent kinase inhibitor roscovitine and the nucleoside analog sangivamycin induce apoptosis in caspase-3 deficient breast cancer cells independent of caspase mediated P-glycoprotein cleavage:implications for therapy of drug resistant breast cancers [J]. Cell Cycle,2009,8(9):1421-1425.
[86]Gibalova L, Seres M, Rusnak A, et al. P-glycoprotein depresses cisplatin sensitivity in L1210 cells by inhibiting cisplatin-induced caspase-3 activation [J]. Toxicol In Vitro,2012,26(3):435-444.
[87]Kaidoh K, Miyauchi S, Abe A, et al. Rhodamine 123 efflux transporter in Haloferax volcanii is induced when cultured under 'metabolic stress' by amino acids:the efflux system resembles that in a doxorubicin-resistant mutant [J]. Biochem J,1996,314 (Pt 1):355-9.
[88]Makin G, Dive C. Recent advances in understanding apoptosis:new therapeutic opportunities in cancer chemotherapy [J]. Trends Mol Med,2003,9(6):251-255.
[89]Liu DL, Bu H, Li H, Chen H, et al. Emodin reverses gemcitabine resistance in pancreatic cancer cells via the mitochondrial apoptosis pathway in vitro [J]. Int J Oncol,2012,40(4):1049-1057.
[90]Makin G, Dive C. Recent advances in understanding apoptosis:new therapeutic opportunities in cancer chemotherapy [J]. Trends Mol Med,2003,9(6):251-255.
[91]Ola MS, Nawaz M, Ahsan H. Role of Bcl-2 family proteins and caspases in the regulation of apoptosis[J]. Mol Cell Biochem.2011,351(1-2):41-58.
[92]Edlich F, Banerjee S, Suzuki M, et al. Bcl-x(L) retrotranslocates Bax from the mitochondria into the cytosol[J]. Cell,2011,145(1):104-16.
[93]陈辉,刘金祥,李洪海,等.大黄素对人胰腺癌细胞株SW1990线粒体膜电位作用的影响[J].中华中医药杂志,2013,28(9)：2610-2613.
[94]王晶,伟忠民.秦皮乙素诱导人肝癌细胞SMMC-7721凋亡的机制研究[J].中成药,2012,34(11)：2059-2063.
[95]Wesarg E, Hoffarth S, Wiewrodt R, et al. Targeting Bcl-2 family proteins to overcome drug resistance in non-small cell lung cancer[J].Int J Cancer,2007,121(11):2387-2394.
[96]Wesarg E, Hoffarth S, Wiewrodt R, et al. Targeting Bcl-2 family proteins to overcome drug resistance in non-small cell lung cancer [J]. Int J Cancer,2007,121(11):2387-2394.
[97]Zhao Y, Zhang GL, Zeng BF, et al. Enhanced chmeosensitivity of drug-resistant osteosarcoma cells by lentivirus-mediatetd Bcl-2 silencing [J]. Biochem Biophys Res Commun,2009,390(3):642-647.
[98]Zaffaroni N, Daidone MG. Survivin expression and resistance to anticancer treatments:perspectives for new therapeutic interventions [J]. Drug Resist Updat,2002,5(2):65-72.
[99]张爱凤,陈平圣,鲁勤,等TUNEL法原位检测肿瘤组织细胞凋亡的改进[J].临床与实验病理学杂志,2008,23(5)：628-629.
[100]Zhai BJ, Shao ZY, Zhao CL, et al. Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice[J]. World J Gastroenterol,2006,12(41):6614-6619.
[101]Chen YB, Yan ML, Gong JP, et al. Establishment of hepatocellular carcinoma multidrug resistant monoclone cell line HepG2/mdrl [J]. Chin Med J (Engl),2007,120(8):703-707.
[102]Chen SY, Hu SS, Dong Q, et al. Establishment of paclitaxel-resistant breast cancer cell line and nude mice models, and underlying multidrug resistance mechanisms in vitro and in vivo[J]. Asian Pac J Cancer Prev,2013,4(10):6135-6140.
[103]Si M, Zhao J, Li X, et al. Reversion effects of curcumin on multidrug resistance of MNNG/HOS human osteosarcoma cells in vitro and in vivo through regulation of P-glycoprotein [J]. Chin Med J (Engl), 2013,26(21):4116-4123.
[104]吴细丕,钱林法.实验动物与肿瘤研究[M].北京：中国医药科技出版社,2000：112.
[105]Mimnaugh EG, Fairchild CR, Fruehauf JP, et al. Biochemical and pharmacological characterization of MCF-7 drug-sensitive and AdrR multidrug-resistant human breast tumor xenografts in athymic nude mice[J]. Biochem Pharmacol,1991,42(2):391-402.
[106]王树滨,齐静,刘桂英,等.多药耐药细胞系K562/A02裸小鼠皮下移植瘤模型的建立与鉴定[J].中国肿瘤临床,2006,33(13)：721-724.
[107]高启龙,李寒冰,姚亚民,等.蜂毒素(Mel)对裸鼠骨肉瘤的抑制作用与影响肿瘤血管生成、细胞增殖和凋亡的关系[J].复旦学报(医学版),2012,39(3)：283-288.
[108]Hasei J, Sasaki T, Tazawa H, et al. Dual programmed cell death pathways induced by p53 transactivation overcome resistance to oncolytic adenovirus in human osteosarcoma cells [J]. Mol Cancer Ther,2013,12 (3):314-325.
[1]Schwartz CL, Gorlick R, Teot L, et al. Children's Oncology Group. Multiple drug resistance in osteogenic sarcoma:INTO133 from the Children's Oncology Group [J]. J Clin Oncol,2007,25(15): 2057-2062.
[2]Chou AJ, Gorlick R. Chemotherapy resistance in osteosarcoma:current challenges and future directions [J]. Expert Rev Anticancer Ther,2006,6:75-85.
[3]Dean M, Fojo T, Bateas S. Tumour stem cell and drug resistance [J]. Nat Rev Cancer,2005, 5(4):275-284.
[4]Lourda M, Trouqakos IP, Gonos ES. Development of resistance to chemotherapeutic drugs in human osteosarcoma cell lines largely depends on up-regulation of clusterin/Apolipoprotein [J]. Int J Cancer,2007, 120(3):611-622.
[5]Takara K, Sakaeda T, Okumura K. An update on overcoming MDR1-mediated multidrug resistance in cancer chemotherapy [J]. Curr Pharm Des,2006,12(3):273-286.
[6]黄纲,李佛保,韩士英,等.骨肉瘤组织中多药耐药基因ndr-1 mRNA和P-糖蛋白的表达及其与临床病理特点关系的研究[J].中华骨科杂志,2000,20(1)：21-25.
[7]Gomes CM, van Paassen H, Romeo S, et al. Mutidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA ananlysis and functional radiotracer studies [J]. Nuclear Medicine and Biology,2006,33(7):831-840.
[8]Susa M, Lyer AK, Ryu K, et al.Inhibition of ABCB1 (MDR1) expression by an siRNA nanoparticulate delivery system to overcome drug resistance in osteosarcoma[J]. PLos One,2010,5(5):e10764.
[9]Adams JM, Cory S. The Bcl-2 apoptotic switch in cancer evelopment and therapy [J]. Oncogene,2007, 26(9):1324-1337.
[10]吴小三,张春林,赵曜,等.应用Cycling D1和Bcl-2为靶标的siRNA慢病毒载体构建和鉴定[J].中国组织工程研究与临床康复,2010,14(20)：3709-3713.
[11]Yao Zhao, Chun-lin Zhang, Bing-fang Zeng, et al. Enhanced chemosensitivity of drug-resistant osteosarcoma cells by lentivirus-mediated Bcl-2 silencing [J]. Biochemical and Biophysical Research Communications,2009,390(3):642-647.
[12]聂涛,戴闽,宗世璋.骨肉瘤中Survivin蛋白表达及其与P-糖蛋白、bcl-2蛋白表达的相关性[J].临床骨科杂志,2005,8(3)：211-214.
[13]傅培荣,李书忠,李萍Suivivin、MMP-2、p-53蛋白在骨肉瘤中表达及临床意义[.J].山东医药,2010,50(19)：49-50.
[14]王井伟,田海梅,刘义,等Survivin-siRNA对骨肉瘤MG-63细胞药物敏感性的研究[J].军医进修学院学报,2010,31(1)：72-74.
[15]安贵峰,吕刚,曾艳,等.转染Livin基因后骨肉瘤细胞系的多药耐药性研究[J].现代肿瘤医学,2009,17(4)：609-613.
[16]Huang G, Mills L, Worth LL. Express of human glutathionw S-transferase Pl mediates the chemosensitivity of osteosarcoma cells [J].Mol Cancer Ther,2007,6(5):1610-1619.
[17]Pasello M, Michelacci F, Scionti I, et al.Overcoming Glutathione S-Transferase Pl-Related Cisplatin Resistance in Osteosarcoma [J]. Cancer Res,2008,68(16):6661-6668.
[18]汪毅,周琪.多药耐药相关蛋白1、谷胱甘肽S转移酶和DNA拓扑酶Ⅱ在骨肉瘤中的表达[J].重庆医学,2006,35(14)：1273-1275.
[19]Yu Z, Pestell TG, Lisanti MP, et al. Cancer stem cells [J]. Int J Biochem Cell Biol,2012,44(12): 2144-2151.
[20]Gupta PB, Chaffer CL, Weinberg RA. Cancer stem cell:mirage or reality? [J]. Nat Med,2009, 15(9):1010-1012.
[21]Tang QL, Liang Y, Xie XB, et al. Enrichment of osteosarcoma stem cell by chemotherapy [J]. Chin J Cancer,2011,30(6):426-432.
[22]Mimeault M, Batra SK. Functions of tumorigenic andmigrating cancer progenitor cells in cancer progressionand metastasis and their therapeutic implications [J]. Cancer Metastasis Rev,2007, 26(1):203-214.
[23]Martins-Neves SR, Lopes AO, do Carmo A, et al. Theraputic implications of an enriched cancer stem-like cell population in a human osteosarcoma cell line [J]. BMC Cancer,2012,12:139.
[24]Tirino V, Desiderio V, d'Aquino R, et al. Detection and Characterization of CD 133+ Cancer Stem Cells in Human Solid Tumours [J]. PLoS One,2008,3(10):e3469.
[25]O'Connor R. A review of mechanisms of circumvention and modulation of chemotherapeutic drug resistance [J].Curr Cancer Drug Targets,2009,9(3):273-280.
[26]Minderman H, O'Loughlin KL, Pendyala L, et al. VX-710 (biricodar) increase drug retention and enhances chemosensitivity in resistant cells overexpressing P-glycoprotein,multidrug resistance protein,and breast cancer resistance protein [J].Clin Cancer Res,2004,10:1826-1834.
[27]Duan Z, Brakora KA, Seiden MV. Inhibition of ABCB1 (MDR1) and ABCB4 (MDR3) expressing by small interfering RNA and reversal of paclitaxel resistance in human ovarian cancer cells [J]. Mol Cancer Ther,2004,3(7):833-838.
[28]车彪,邵增务,王凯,等stathmin基因沉默协同增效多系紫杉醇对骨肉瘤细胞化疗作用的实验研究[J].中国临床研究,2010,23(3)：181-183.
[29]史曦凯,张翼军,赵春景.人参皂甙单体Rb1对多药耐药细胞K562/HHT的耐药逆转作用[J].第三军医大学学报,1999,21(11)：825-827.
[30]田晖,谭启超.双苄基异喹啉类生物碱与小蘖胺逆转多药抗药性的比较研究[J].药学学报,1997,32(4)：245-250.
[31]王进,沈靖南.粉防己碱逆转骨肉瘤细胞系多药耐药性的研究[J].实用临床医学,2005,6(7)：7-10.
[32]肖扬,王万春.姜黄素对人骨肉瘤细胞株/阿霉素多药耐药逆转作用的实验研究[J].2011,40(1)：103-106.
[33]石宇雄,黄永明,许少健,等.参麦注射液对人成骨肉瘤多药耐药细胞株R-OS-732逆转作用[J].广东医学,2006,27(8)：1200-1201.
[34]石宇雄,黄永明,张娴,等.骨癌通泰方对人成骨肉瘤细胞R-KHOS MRP基因表达的影响[J].陕西中医,2009,29(11)：1552-1553.
[2]Mirabello L, Troisi RJ and Savage SA. Osteosarcoma incidence and survival rates from 1973 to 2004: data from the Surveillance, Epidemiology, and End Results Program [J]. Cancer,2009,115 (7):1531-1543.
[3]Rajani R and Gibbs CP. Treatment of bone tumors [J]. Surg Pathol Clin,2012,5 (1):301-318.
[4]Ando K, Heymann MF, Stresing V, et al. Current therapeutic strategies and novel approaches in osteosarcoma [J].Cancers,2013,5(2):591-616.
[5]Chou AJ, Gorlick R. Chemotherapy resistance in osteosarcoma:current challenges and future directions [J]. Expert Rev Anticancer Ther,2006,6(7):1075-1085.
[6]Schwartz CL, Gorlick R, Teot L, et al. Children's Oncology Group:Multiple drug resistance in osteogenic sarcoma:INTO133 from the Children's Oncology Group [J]. J Clin Oncol,2007,25(15): 2057-2062.
[7]Baguley BC. Multiple drug resistance mechanisms in cancer [J]. Mol Biotechnol,2010,46(3):308-316.
[8]Liu FS. Mechanisms of chemotherapeutic drug resistance in cancer therapy--a quick review [J]. Taiwan J Obstet Gynecol,2009,48(3):239-44.
[9]Shukla S, Ohnuma S, Ambudkar SV. Improving cancer chemotherapy with modulators of ABC drug transporters [J]. Curr Drug Targets,2011,12(5):621-630.
[10]杨华,黄文荣,于力.新型抗肿瘤药物的临床应用及研究进展[J].中国新药杂志,2010,19(6)：480-485.
[11]Li-Weber M. Targeting apoptosis pathways in cancer by Chinese medicine [J]. Cancer Lett,2013, 332(2):304-312.
[12]Yang L, Wei DD, Chen Z, et al. Reversal effects of traditional Chinese herbs on multidrug resistance in cancer cells [J]. Nat Prod Res,2011,25(19):1885-1889.
[13]王辉,孙桂芝.孙桂芝治疗骨肉瘤经验[J].世界中医药,2012,7(1)：21-22.
[14]孟丽萍,顾雪峰.片仔癀联合治疗毒热瘀结型原发性肝癌免疫指标变化的临床观察[J].九江医学,2008,23(1)：31-37.
[15]林明和,朱德增.片仔癀联合化疗治疗热毒瘀结型晚期结肠癌临床研究[J].福建中医药,2102,43(1)：8-9.
[16]Lu L, Wai MS, Yew DT, et al. Pien Tze Huang, a composite Chinese traditional herbal extract, affects survival of neuroblastoma cells [J]. Int J Neurosci,2009,119(2):255-262.
[17]Lin Jiu-mao, Wei Li-hui, Chen You-qin, et al. Pien Tze Huang-induced apoptosis in human colon cancer HT-29 cells is associated with regulation the Bcl-2 family and activation of caspase3 [J].Chin J Integr Med,2011, 11(9):685-690.
[18]Shen A, Hong F, Liu L, et al. Pien Tze Huang inhibits the proliferation of human colon carcinoma cells by arresting Gl/S cell cycle progression[J]. Oncol Lett.2012,4(4):767-770.
[19]Shen A, Chen Y, Hong F, et al, Pien Tze Huang suppresses IL-6-inducible STAT3 activation in human colon carcinoma cells through induction of SOCS3 [J]. Oncol Rep.2012,28(6):2125-30.
[20]张俐,曾启清,林建华.P27基因联合中药片仔癀对人骨肉瘤抑制作用观察[J].中华中医药杂志,2009,24(4)：511-515.
[21]张俐,于波,林建华.中药片仔癀胶囊对骨肉瘤细胞诱导凋亡的作用[J].中国骨伤,2009, 22(4):265-268.
[22]张燕,王琪鸿,张俐,等.片仔癀对人骨肉瘤细胞U20S细胞周期的影响[J].江西中医学院学报,2012,24(5)：61-63.
[23]杨德建.片仔癀对裸鼠原位移植骨肉瘤的作用及机制研究[D],福建：福建中医药大学,2013.
[24]Wang C, Guo LB, Ma JY, et al. Establishment and characterization of a paclitaxel-resistant human esophageal carcinoma cell line [J]. Int J Oncol,2013,43(5):1607-1617.
[25]Zhang X, Yashiro M, Qiu H, et al. Establishment and characterization of multidrug-resistant gastric cancer cell lines [J]. Anticancer Res,2010,30(3):915-921.
[26]赵斌,葛金芳,朱娟娟,等小议在MTT法测细胞增殖抑制率中IC50的计算方法[J].安徽医药,2007,11(9)：834-835.
[27]Ando K, Heymann MF, Stresing V, et al. Current therapeutic strategies and novel approaches in osteosarcoma [J]. Cancers (Basel),2013,5(2):591-616.
[28]Scheetze SM. Chemotherapy in the management of osteosarcoma and E wing's sarcoma [J]. J Natl Compr Canc Netw,2007,5(4):449-455.
[29]Saraswathy M, Gong S. Different strategies to overcome multidrug resistance in cancer [J]. Biotechnol Adv,2013,31(8):1397-1407.
[30]Niu BH, Wang JJ, Xi Y, et al. The establishment and characterization of adriamycin-resistant cell lines derived from Saos-2 [J]. Medical Science Monitor,2010,16(6):BR184-192.
[31]Uchibori K, Kasamatsu A, Sunaga M, et al. Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines [J]. Int J Oncol,2012,40(4):1005-1010.
[32]Ahrens AK, Chaturvedi NK, Nordgren TM, et al. Establishment and characterization of therapy-resistant mantle cell lymphoma cell lines derived from different tissue sites [J]. Leuk Lymphoma, 2012,53(11):2269-2278.
[33]Carrle D, Bielack SS. Current strategies of chemotherapy in osteosarcoma [J]. Int Orthop,2006, 30(6):445-451.
[34]Rebbaa A. Targeting senescence pathways to reverse drug resistance in cancer [J]. Cancer Lett,2005, 219(1):1-13.
[35]Saretzki G.Cellular senescence in the development and treatment of cancer [J]. Curr Pharm Des,2010, 16(1):79-100.
[36]Juliao RL and Linvg V. A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants [J]. Biochim Biophys Acta,1976,455:152-162.
[37]Binkhathlan Z and Lavasanifar A. P-glycoprotein inhibition as a therapeutic approach for overcoming multidrug resistance in cancer:current status and future perspectives [J]. Curr Cancer Drug Targets,2013, 13(3):326-346.
[38]Mueller CF, Wassmann K, Widder JD, et al. Multidrug resistance protein-1 affects oxidative stress, endothelial dysfunction, and atherogenesis via leukotriene C4 export [J]. Circulation,2008, 117(22):2912-2918.
[39]Johnstone RW, Ruefli AA, Tainton KM, et al. A role for P-glycoprotein in regulating cell death [J]. Leuk Lymphoma,2000,38(1-2):1-11.
[40]Yamagishi T, Sahni S, Sharp DM, et al. P-glycoprotein mediates drug resistance via a novel mechanism involving lysosomal sequestration [J]. J Biol Chem,2013,288(44):31761-31771.
[41]Gomes CM, van Paassen H, Romeo S, et al. Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA analysis and functional radiotracer studies [J]. Nuclear Medicine and Biology,2006,33(7):831-840.
[42]Cole SP, Bhardwai G, Gerlach JH, et al. Over-expression of a transporters gene in a multidrug-resistant human lung cancer cell line [J]. Science,1992,5088:1650-1654.
[43]Leslie EM, Deeley RG, Cole SP. Multidrug resistance proteins:role of P-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense [J]. Toxicol Appl Pharmacol,2005,204(3):216-237.
[44]Keppler D. Multidrug resistance proteins (MRPs, ABCCs):importance for pathophysiology and drug therapy [J]. Handb Exp Pharmacol,2011,201:299-323.
[45]Homolya L, Varadi A and Sarkadi B. Multidrug resistance-associated proteins:Export pumps for conjugates with glutathione, glucuronate or sulfate [J]. Biofactors,2003,17(1-4):103-114.
[46]Wang L, Zhang GM, Feng Zh. Down-regulation of survivin expression reversed multidrug resistance in adriamycin-resistant HL-60/ADR cell line[J]. Acta Pharmacol Sin,2003,24(12):1235-1240.
[47]Ambrosini G, Adida C, Altieri DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med,1997,3(8):917-921.
[48]Shoeneman JK, Ehrhart EJ 3rd, Eickhoff JC, et al. Expression and function of survivin in canine osteosarcoma [J]. Cancer Res,2012,72(1):249-259.
[49]Zou J, Gan M, Mao N, et al. Sensitization of osteosarcoma cell line SaOS-2 to chemotherapy by downregulating survivin [J]. Arch Med Res,2010,41(3):162-9.
[50]黄芳.植物多酚类化合物联合阿霉素抑制MCF-7/ADM细胞的筛选及白藜芦醇逆转耐药性作用研究[D],福建：福建医科大学,2009：34-35.
[51]Chou AJ, Geller DS, Gorlick R. Therapy for osteosarcoma:where do we go from here? [J]. Paediatr Drugs,2008,10(5):315-327.
[52]Ayan I, Kebudi R, Ozger H. Childhood osteosarcoma:multimodal therapy in a single-institution Turkish series [J]. Cancer Treat Res.2009,152:319-338.
[53]牛晓辉,徐海荣.骨肉瘤的化疗进展[J].中国癌症杂志,2012,22(9)：663-669.
[54]凌昌全.恶性肿瘤病因病机之我见[J].中医杂志,2009,50(10)：952-953.
[55]吴薏婷,吴锦燕.陈庆强主任治疗恶性肿瘤经验[J].辽宁中医药大学学报,2011,13(5)：164-165.
[56]潘万刚.中药三棱祁甲汤治疗骨肉瘤22例临床疗效分析[J].中华临床医学杂志,2005,6(9)：88-90.
[57]邹存华,付婷婷,鲁强,等.三七总皂苷抑制Hela细胞增殖的实验研究[J].中华肿瘤防治杂志2012,19(12)：888-891.
[58]杨茹萍,陈彤,陈亚娟,等.三七提取物的体外抗肿瘤药理作用及其成分分析[J].昆明医学院学报,2011,9：4-6.
[59]Wang CZ, Luo XJ, Zhang B, et al. Notoginseng enhances anticancer effect of 5-fluotouiacill on human colorectal cancer cells[J]. Cancer Chemother Pharmacol,2007,60(1):69-79.
[60]黄芳.牛黄对胃癌细胞BCG-823增殖抑制、诱导凋亡的实验研究[D].湖南：中南大学,2013：29.
[61]吴志远,李芳芳,张昆,等.人工牛黄对小鼠乳腺癌肺转移的影响[J].中草药,2012,43(10)：2013-2016.
[62]多明荷,孙莉娜,应森林,等.西黄丸通过ERK/MAPK信号通路对人结肠癌裸鼠移植瘤的影响[J].中华中医药杂志,2013,28(10)：3055-3058.
[63]Massart C, Gibassier J, Raoul M, et al. Cyclosporin A, verapamil and S9788 reverse doxorubicin resistance in a human medullary thyroid carcinoma cell line [J]. Anticancer Drugs,1995, 6(1):135-46.
[64]O'Brien MM, Lacayo NJ, Lum BL,et al. Phase I study of valspodar (PSC-833) with mitoxantrone and etoposide in refractory and relapsed pediatric acute leukemia:a report from the Children's Oncology Group p [J]. Pediatr Blood Cancer,2010,54(5):694-702.
[65]Bottova I, Sauder U, Olivieri V, et al. The P-glycoprotein inhibitor GF120918 modulates Ca2+-dependent processes and lipid metabolism in Toxoplasma gondii [J]. PLoS One,2010,5(4):el0062.
[66]Callies S, de Alwis DP, Harris A, et al. A population pharmacokinetic model for paclitaxel in the presence of a novel P-gp modulator, Zosuquidar Trihydrochloride (LY335979) [J].Br J Clin Pharmacol,2003,56(1):46-56.
[67]Jia L, Ma S, Hou X, et al. The synergistic effects of traditional Chinese herbs and radiotherapy for cancer treatment [J]. Oncol Lett,2013,5(5):1439-1447.
[68]Wu G, Chu J, Huang Z, et al. Xiao Jin Wan, a traditional Chinese herbal formula, inhibits proliferation via arresting cell cycle progression at the G2/M phase and promoting apoptosis via activating the mitochondrial-dependent pathway in U-2OS human osteosarcoma cells [J]. Int J Oncol Mar,2013,42(3): 1070-1080.
[69]吕仙梅,郑坚,朱莹杰,等.中医药联合化疗对大肠癌、期患者生存期的影响[J].中国中西结合杂志,2012,32(9)：1166-1170.
[70]余文燕,王国娟,许建华,等.肠胃清颗粒协同卡铂对人结肠癌耐药细胞增殖及凋亡的影响[J].中医杂志,2013,54(21)：1854-1858.
[71]Deng S, Hu B, An HM, et al. Teng-Long-Bu-Zhong-Tang, a Chinese herbal formula, enhances anticancer effects of 5--Fluorouracil in CT26 colon carcinoma [J]. BMC Complement Altern Med,2013, 13:128.
[72]肖扬,王万春.姜黄素对人骨肉瘤细胞株/阿霉素多药耐药逆转作用的实验研究[J].卫生研究,2011,40(1)103-106.
[73]Yang L, Wei DD, Chen Z, et al. Reversal of multidrug resistance in human breast cancer cells by Curcuma wenyujin and Chrysanthemum indicum [J]. Phytomedicine,2011,18(8-9):710-718.
[74]Zhu X, Sui M, Fan W. In vitro and in vivo characterizations of tetrandrine on the reversal of P-glycoprotein-mediated drug resistance to paclitaxel [J]. Anticancer Res.2005,25(3B):1953-1962.
[75]王进,沈靖南.粉防己碱逆转骨肉瘤细胞系多药耐药性的研究[J].实用临床医学,2005,6(7)：7-10.
[76]Xie X, Yin J, Jia Q, Wang J, et al. Quercetin induces apoptosis in the methotrexate-resistant osteosarcoma cell line U2-OS/MTX300 via mitochondrial dysfunction and dephosphorylation of Akt [J]. Oncol Rep,2011,26(3):687-693.
[77]余文燕,孙珏,徐建华,等.肠胃清药物血清协同奥沙利铂对人结肠癌耐药细胞增殖及凋亡的影响[J].中国实验方剂学杂志,,2013,19(10)：182-186.
[78]聂昊,王晖.P糖蛋白在不同组织中的分布与功能研究进展[J],广东药学院学报,2012,28(4)：456-460.
[79]Leslie EM, Deeley RG, Cole SP, et al. Multidrug resistance proteins:role of P-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense [J], Toxicol Appl Pharmacol.2005,204(3):216-237.
[80]Gomes CM, van Paassen H, Romeo S, et al. Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA analysis and functional radiotracer studies [J]. Nucl Med Biol,2006,33(7): 831-840.
[81]Serra M, Scotlandi K, Reverter-Branchat G, et al. Value of P-Glycoprotein and Clinicopathologic Factors as the Basis for New Treatment Strategies in High-Grade Osteosarcoma of the Extremities [J]. J Clin Oncol,2003,21(3):536-542.
[82]Sauna ZE, Ambudkar SV. About a switch:how P-glycoprotein (ABCB1) harnesses the energy of ATP binding and hydrolysis to do mechanical work [J]. Mol Cancer Ther,2007,6(1):13-23.
[83]Rosenberg MF, Kamis AB, Callaghan R, et al. Three-dimensional structures of the mammalian multidrug resistance P-glycoprotein demonstrate major conformational changes in the transmembrane domains upon nucleotide binding [J]. J Biol Chem,2003,278(10):8294-8299.
[84]Johnstone RW, Ruefli AA, Tainton KM, et al. A role for P-glycoprotein in regulating cell death [J]. Leuk Lymphoma,2000,38(1-2):1-11.
[85]Cappellini A, Chiarini F, Ognibene A, et al. The cyclin-dependent kinase inhibitor roscovitine and the nucleoside analog sangivamycin induce apoptosis in caspase-3 deficient breast cancer cells independent of caspase mediated P-glycoprotein cleavage:implications for therapy of drug resistant breast cancers [J]. Cell Cycle,2009,8(9):1421-1425.
[86]Gibalova L, Seres M, Rusnak A, et al. P-glycoprotein depresses cisplatin sensitivity in L1210 cells by inhibiting cisplatin-induced caspase-3 activation [J]. Toxicol In Vitro,2012,26(3):435-444.
[87]Kaidoh K, Miyauchi S, Abe A, et al. Rhodamine 123 efflux transporter in Haloferax volcanii is induced when cultured under 'metabolic stress' by amino acids:the efflux system resembles that in a doxorubicin-resistant mutant [J]. Biochem J,1996,314 (Pt 1):355-9.
[88]Makin G, Dive C. Recent advances in understanding apoptosis:new therapeutic opportunities in cancer chemotherapy [J]. Trends Mol Med,2003,9(6):251-255.
[89]Liu DL, Bu H, Li H, Chen H, et al. Emodin reverses gemcitabine resistance in pancreatic cancer cells via the mitochondrial apoptosis pathway in vitro [J]. Int J Oncol,2012,40(4):1049-1057.
[90]Makin G, Dive C. Recent advances in understanding apoptosis:new therapeutic opportunities in cancer chemotherapy [J]. Trends Mol Med,2003,9(6):251-255.
[91]Ola MS, Nawaz M, Ahsan H. Role of Bcl-2 family proteins and caspases in the regulation of apoptosis[J]. Mol Cell Biochem.2011,351(1-2):41-58.
[92]Edlich F, Banerjee S, Suzuki M, et al. Bcl-x(L) retrotranslocates Bax from the mitochondria into the cytosol[J]. Cell,2011,145(1):104-16.
[93]陈辉,刘金祥,李洪海,等.大黄素对人胰腺癌细胞株SW1990线粒体膜电位作用的影响[J].中华中医药杂志,2013,28(9)：2610-2613.
[94]王晶,伟忠民.秦皮乙素诱导人肝癌细胞SMMC-7721凋亡的机制研究[J].中成药,2012,34(11)：2059-2063.
[95]Wesarg E, Hoffarth S, Wiewrodt R, et al. Targeting Bcl-2 family proteins to overcome drug resistance in non-small cell lung cancer[J].Int J Cancer,2007,121(11):2387-2394.
[96]Wesarg E, Hoffarth S, Wiewrodt R, et al. Targeting Bcl-2 family proteins to overcome drug resistance in non-small cell lung cancer [J]. Int J Cancer,2007,121(11):2387-2394.
[97]Zhao Y, Zhang GL, Zeng BF, et al. Enhanced chmeosensitivity of drug-resistant osteosarcoma cells by lentivirus-mediatetd Bcl-2 silencing [J]. Biochem Biophys Res Commun,2009,390(3):642-647.
[98]Zaffaroni N, Daidone MG. Survivin expression and resistance to anticancer treatments:perspectives for new therapeutic interventions [J]. Drug Resist Updat,2002,5(2):65-72.
[99]张爱凤,陈平圣,鲁勤,等TUNEL法原位检测肿瘤组织细胞凋亡的改进[J].临床与实验病理学杂志,2008,23(5)：628-629.
[100]Zhai BJ, Shao ZY, Zhao CL, et al. Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice[J]. World J Gastroenterol,2006,12(41):6614-6619.
[101]Chen YB, Yan ML, Gong JP, et al. Establishment of hepatocellular carcinoma multidrug resistant monoclone cell line HepG2/mdrl [J]. Chin Med J (Engl),2007,120(8):703-707.
[102]Chen SY, Hu SS, Dong Q, et al. Establishment of paclitaxel-resistant breast cancer cell line and nude mice models, and underlying multidrug resistance mechanisms in vitro and in vivo[J]. Asian Pac J Cancer Prev,2013,4(10):6135-6140.
[103]Si M, Zhao J, Li X, et al. Reversion effects of curcumin on multidrug resistance of MNNG/HOS human osteosarcoma cells in vitro and in vivo through regulation of P-glycoprotein [J]. Chin Med J (Engl), 2013,26(21):4116-4123.
[104]吴细丕,钱林法.实验动物与肿瘤研究[M].北京：中国医药科技出版社,2000：112.
[105]Mimnaugh EG, Fairchild CR, Fruehauf JP, et al. Biochemical and pharmacological characterization of MCF-7 drug-sensitive and AdrR multidrug-resistant human breast tumor xenografts in athymic nude mice[J]. Biochem Pharmacol,1991,42(2):391-402.
[106]王树滨,齐静,刘桂英,等.多药耐药细胞系K562/A02裸小鼠皮下移植瘤模型的建立与鉴定[J].中国肿瘤临床,2006,33(13)：721-724.
[107]高启龙,李寒冰,姚亚民,等.蜂毒素(Mel)对裸鼠骨肉瘤的抑制作用与影响肿瘤血管生成、细胞增殖和凋亡的关系[J].复旦学报(医学版),2012,39(3)：283-288.
[108]Hasei J, Sasaki T, Tazawa H, et al. Dual programmed cell death pathways induced by p53 transactivation overcome resistance to oncolytic adenovirus in human osteosarcoma cells [J]. Mol Cancer Ther,2013,12 (3):314-325.
[1]Schwartz CL, Gorlick R, Teot L, et al. Children's Oncology Group. Multiple drug resistance in osteogenic sarcoma:INTO133 from the Children's Oncology Group [J]. J Clin Oncol,2007,25(15): 2057-2062.
[2]Chou AJ, Gorlick R. Chemotherapy resistance in osteosarcoma:current challenges and future directions [J]. Expert Rev Anticancer Ther,2006,6:75-85.
[3]Dean M, Fojo T, Bateas S. Tumour stem cell and drug resistance [J]. Nat Rev Cancer,2005, 5(4):275-284.
[4]Lourda M, Trouqakos IP, Gonos ES. Development of resistance to chemotherapeutic drugs in human osteosarcoma cell lines largely depends on up-regulation of clusterin/Apolipoprotein [J]. Int J Cancer,2007, 120(3):611-622.
[5]Takara K, Sakaeda T, Okumura K. An update on overcoming MDR1-mediated multidrug resistance in cancer chemotherapy [J]. Curr Pharm Des,2006,12(3):273-286.
[6]黄纲,李佛保,韩士英,等.骨肉瘤组织中多药耐药基因ndr-1 mRNA和P-糖蛋白的表达及其与临床病理特点关系的研究[J].中华骨科杂志,2000,20(1)：21-25.
[7]Gomes CM, van Paassen H, Romeo S, et al. Mutidrug resistance mediated by ABC transporters in osteosarcoma cell lines:mRNA ananlysis and functional radiotracer studies [J]. Nuclear Medicine and Biology,2006,33(7):831-840.
[8]Susa M, Lyer AK, Ryu K, et al.Inhibition of ABCB1 (MDR1) expression by an siRNA nanoparticulate delivery system to overcome drug resistance in osteosarcoma[J]. PLos One,2010,5(5):e10764.
[9]Adams JM, Cory S. The Bcl-2 apoptotic switch in cancer evelopment and therapy [J]. Oncogene,2007, 26(9):1324-1337.
[10]吴小三,张春林,赵曜,等.应用Cycling D1和Bcl-2为靶标的siRNA慢病毒载体构建和鉴定[J].中国组织工程研究与临床康复,2010,14(20)：3709-3713.
[11]Yao Zhao, Chun-lin Zhang, Bing-fang Zeng, et al. Enhanced chemosensitivity of drug-resistant osteosarcoma cells by lentivirus-mediated Bcl-2 silencing [J]. Biochemical and Biophysical Research Communications,2009,390(3):642-647.
[12]聂涛,戴闽,宗世璋.骨肉瘤中Survivin蛋白表达及其与P-糖蛋白、bcl-2蛋白表达的相关性[J].临床骨科杂志,2005,8(3)：211-214.
[13]傅培荣,李书忠,李萍Suivivin、MMP-2、p-53蛋白在骨肉瘤中表达及临床意义[.J].山东医药,2010,50(19)：49-50.
[14]王井伟,田海梅,刘义,等Survivin-siRNA对骨肉瘤MG-63细胞药物敏感性的研究[J].军医进修学院学报,2010,31(1)：72-74.
[15]安贵峰,吕刚,曾艳,等.转染Livin基因后骨肉瘤细胞系的多药耐药性研究[J].现代肿瘤医学,2009,17(4)：609-613.
[16]Huang G, Mills L, Worth LL. Express of human glutathionw S-transferase Pl mediates the chemosensitivity of osteosarcoma cells [J].Mol Cancer Ther,2007,6(5):1610-1619.
[17]Pasello M, Michelacci F, Scionti I, et al.Overcoming Glutathione S-Transferase Pl-Related Cisplatin Resistance in Osteosarcoma [J]. Cancer Res,2008,68(16):6661-6668.
[18]汪毅,周琪.多药耐药相关蛋白1、谷胱甘肽S转移酶和DNA拓扑酶Ⅱ在骨肉瘤中的表达[J].重庆医学,2006,35(14)：1273-1275.
[19]Yu Z, Pestell TG, Lisanti MP, et al. Cancer stem cells [J]. Int J Biochem Cell Biol,2012,44(12): 2144-2151.
[20]Gupta PB, Chaffer CL, Weinberg RA. Cancer stem cell:mirage or reality? [J]. Nat Med,2009, 15(9):1010-1012.
[21]Tang QL, Liang Y, Xie XB, et al. Enrichment of osteosarcoma stem cell by chemotherapy [J]. Chin J Cancer,2011,30(6):426-432.
[22]Mimeault M, Batra SK. Functions of tumorigenic andmigrating cancer progenitor cells in cancer progressionand metastasis and their therapeutic implications [J]. Cancer Metastasis Rev,2007, 26(1):203-214.
[23]Martins-Neves SR, Lopes AO, do Carmo A, et al. Theraputic implications of an enriched cancer stem-like cell population in a human osteosarcoma cell line [J]. BMC Cancer,2012,12:139.
[24]Tirino V, Desiderio V, d'Aquino R, et al. Detection and Characterization of CD 133+ Cancer Stem Cells in Human Solid Tumours [J]. PLoS One,2008,3(10):e3469.
[25]O'Connor R. A review of mechanisms of circumvention and modulation of chemotherapeutic drug resistance [J].Curr Cancer Drug Targets,2009,9(3):273-280.
[26]Minderman H, O'Loughlin KL, Pendyala L, et al. VX-710 (biricodar) increase drug retention and enhances chemosensitivity in resistant cells overexpressing P-glycoprotein,multidrug resistance protein,and breast cancer resistance protein [J].Clin Cancer Res,2004,10:1826-1834.
[27]Duan Z, Brakora KA, Seiden MV. Inhibition of ABCB1 (MDR1) and ABCB4 (MDR3) expressing by small interfering RNA and reversal of paclitaxel resistance in human ovarian cancer cells [J]. Mol Cancer Ther,2004,3(7):833-838.
[28]车彪,邵增务,王凯,等stathmin基因沉默协同增效多系紫杉醇对骨肉瘤细胞化疗作用的实验研究[J].中国临床研究,2010,23(3)：181-183.
[29]史曦凯,张翼军,赵春景.人参皂甙单体Rb1对多药耐药细胞K562/HHT的耐药逆转作用[J].第三军医大学学报,1999,21(11)：825-827.
[30]田晖,谭启超.双苄基异喹啉类生物碱与小蘖胺逆转多药抗药性的比较研究[J].药学学报,1997,32(4)：245-250.
[31]王进,沈靖南.粉防己碱逆转骨肉瘤细胞系多药耐药性的研究[J].实用临床医学,2005,6(7)：7-10.
[32]肖扬,王万春.姜黄素对人骨肉瘤细胞株/阿霉素多药耐药逆转作用的实验研究[J].2011,40(1)：103-106.
[33]石宇雄,黄永明,许少健,等.参麦注射液对人成骨肉瘤多药耐药细胞株R-OS-732逆转作用[J].广东医学,2006,27(8)：1200-1201.
[34]石宇雄,黄永明,张娴,等.骨癌通泰方对人成骨肉瘤细胞R-KHOS MRP基因表达的影响[J].陕西中医,2009,29(11)：1552-1553.
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