束骨姜黄醇对舌癌Tca8113细胞系增殖、凋亡的影响及其相关机制研究
|
摘要
目的:舌癌是口腔颌面部最为多见的恶性肿瘤,近年来在世界范围内其发病率逐年升高,并且其发病年龄越来越小,同时其恶性程度也越来越高。容易发生早期颈部淋巴结转移,且转移率较高为舌癌的主要特点之一。目前舌癌的首选治疗方式是手术治疗,同时,术前、术后常需配合化疗以提高疗效,但是传统的化疗通常存在着肿瘤细胞耐药、化疗药物毒副作用大患者难以耐受等影响治疗效果的不利因素,所以研究和开发治疗恶性肿瘤的新药,成为恶性肿瘤治疗领域研究的重点。目前,国内外学者越来越重视从动植物、矿物质、海洋生物等天然物质中寻找高效低毒的治疗恶性肿瘤的药物,并已取得一定成绩。束骨姜黄醇(Xanthorrhizol)是从姜黄属植物印尼莪术(C.xanthorrhiza)根茎的挥发油中提取出来的一种主要成份,目前研究显示,束骨姜黄醇对宫颈癌、乳腺癌等恶性肿瘤有明显的抑制作用。本课题通过体外培养舌癌Tca8113细胞,研究束骨姜黄醇对舌癌Tca8113细胞增殖及凋亡的影响,探讨束骨姜黄醇在舌癌治疗中的作用机制,为舌癌的化学治疗提供实验依据。
方法:将复苏后的舌癌Tca8113细胞置于37℃,CO_2饱和湿度5%的常规条件下培养,待细胞进入对数生长期后用于实验。
1流式细胞术(FCM)检测细胞凋亡、细胞周期分布:收集不同浓度束骨姜黄醇作用0h、12h、24h、48h、72h的舌癌Tca8113细胞及对照组细胞,冷PBS漂洗。预冷70%乙醇4℃固定,上机检测前离心去除固定液,用0.5%胃蛋白酶消化,碘化丙锭(ProPidium Iodide,PI:50mg/L Trinton X-100 1.0%)室温避光染色。流式细胞仪上机检测,应用软件进行分析。
2流式细胞术(FCM)检测Bcl-2、Bax基因蛋白的表达情况:收集经不同浓度束骨姜黄醇作用0h、12h、24h、48h、72h的舌癌Tca8113细胞及对照组细胞,按常规方法进行荧光标记,流式细胞仪检测荧光强度,以平均荧光强度道数表示蛋白的表达量。
结果:
1束骨姜黄醇对舌癌Tca8113细胞周期的影响。
1.1 20μmol/L、40μmol/L、60μmol/L束骨姜黄醇分别作用于舌癌Tca8113细胞12h后,各用药组S+G2/M期细胞周期百分比与对照组的相比,差异具有显著性(P<0.01);药物作用24h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.05);药物作用48h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.05);药物作用72h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.01)。各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.05)。不同时间点各用药物组间比较,除用药72h后,20μmol/L组和40μmol/L组之间差异无显著性外(P﹥0.05),其余各时间点各组比较差异均具有显著性(P<0.05)。
1.2各用药浓度组作用不同时间S+G2/M期细胞周期百分比的方差分析表明,20μmol/L组中,48h与72h两时间点之间比较,差异无显著性(P﹥0.05),其余各时间点之间,差异具有显著性(P<0.05);40μmol/L组中,48h与72h两时间点之间比较,差异无显著性(P﹥0.05),其余各时间点之间,差异具有显著性(P<0.05);60μmol/L组中,各时间点之间比较,差异均具有显著性(P<0.05)。
1.3经不同浓度药物作用不同时间后,G2/M期细胞周期百分比变化无明显规律性。
2流式细胞仪检测束骨姜黄醇对舌癌Tca8113细胞细胞凋亡的影响。
2.1 20μmol/L、40μmol/L、60μmol/L束骨姜黄醇作用于舌癌Tca8113细胞12h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用24h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用48h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用72h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.01)。不同时间点各用药组间比较,除用药12h后,20μmol/L组和40μmol/L组之间差异无显著性外(P﹥0.05),其余个时间点各组比较差异均具有显著性(P<0.01)。
2.2各用药浓度组作用不同时间细胞凋亡的方差分析表明,除20μmol/L组内48h与72h之间比较,差异无显著性外(P﹥0.05),其余各组中各时间点间比较,差异均具有显著性(P<0.05)。
3 FCM检测束骨姜黄醇作用于舌癌Tca8113细胞后凋亡相关基因蛋白Bcl-2、Bax的表达。
3.1 Bcl-2蛋白表达量:
3.1.1 12h、24h组蛋白表达分别与对照组相比,除20μmol/L组外,其余各组差异具有显著性(P<0.05);48h组蛋白表达与对照组相比,差异具有显著性(P<0.05);72h组蛋白表达与对照组相比,差异具有显著性(P<0.05);各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.05);不同时间点各用药组之间比较,差异具有显著性(P<0.05)。
3.1.2各用药浓度组作用不同时间蛋白表达的方差分析表明,20μmol/L组内各时间点之间差异不显著(P﹥0.05),40μmol/L组中除48h与72h之间差异无显著性(P﹥0.05)外,其余各时间点间差异均显著(P<0.05),60μmol/L组中各时间点间差异均显著(P<0.05)。
3.2 Bax蛋白表达量:各用药浓度组作用不同时间后,各组Bax蛋白表达量经析因设计的方差分析表明,时间因素与剂量因素在束骨姜黄醇对细胞Bax蛋白表达量的影响中,不存在交互效应(P﹥0.05);逐一分析药物作用后,不同时间和不同药物浓度对Bax蛋白表达量的影响,结果表明实验组与对照组的差异不具备显著性(P﹥0.05)。
结论:
1束骨姜黄醇能够显著影响舌癌Tca8113细胞的细胞周期,使细胞周期阻滞在G0/G1期,从而达到抗肿瘤的目的。该药物对细胞周期的阻滞作用呈时间和剂量依赖关系。
2束骨姜黄醇在一定浓度范围内能够呈时间、浓度依赖性地诱导舌癌Tca8113细胞凋亡。
3束骨姜黄醇能够显著影响凋亡相关基因蛋白Bcl-2的表达,使Bcl-2蛋白表达逐渐降低,蛋白表达含量的改变呈剂量依赖性,同时该药物对Bcl-2蛋白表达的影响在达到一定浓度后出现时间依赖性;药物对凋亡相关基因蛋白Bax的表达无明显影响。
Objective: Tongue Carcinoma is the most common malignancy In the oral and maxillofacial careinoma ,in recent years, worldwide ,its incidence is increasing year by year, and its age of onset is smaller and smaller, while the degree of its malignancy is increasing. Prone to early cervical lymph node metastases, and higher transfer rate is one of the main features of tongue Carcinoma. The current treatment of choice for tongue Carcinoma is surgery, while preoperative and postoperative chemotherapy often need to meet in order to improve efficacy, but there is often a traditional chemotherapy drug resistance of tumor cells, chemotherapy drugs in patients with toxic side effects of treatment difficult to tolerate, etc. Effects of unfavorable factors, so the research and development of new drugs to treat cancer is important in a cancer therapeutic area research. Currently, scholars more and more pay attention to find some efficiency and low toxicity of the treatment of cancer drugs from plants , animals, minerals, marine and other natural substances, and has achieved considerable success. Xanthorrhizol is one of the main ingredients extracted from volatile oil of the rhizome of Curcuma xanthorrhiza,the present study shows that Xanthorrhizol can significantly inhibit the Cervical cancer, breast cancer and other malignant tumors .The subject , tongue cancer cell Line Tca8113 by in vitro ,study the Influence of Xanthorrhizol on Tca8113 cell proliferation and apoptosis, discuss the mechanism of action for tongue Carcinoma, to provide experimental basis for the chemical treatment .
Methods: After recovery, Tca8113 cells were 37℃, CO_2 saturated 5% humidity, cultured under routine conditions, until after the cells entered the logarithmic growth phase for experiments.
1 Flow cytometry (FCM) to detect apoptosis, cell cycle distribution: collect control cells and Tca8113 cells which were effected by different concentrations of Xanthorrhizol in 0h, 12h, 24h, 48h, 72h, cold PBS rinse. 4℃70% ethanol pre-cooled fixed, centrifuged to remove pre-test on the machine stationary phase, with 0.5% pepsin digestion, propidium iodide (Propidium Iodide, PI: 50mg / L Trinton X-100 1.0%) at room temperature away from light staining . Detected by flow cytometry on the machine, application software for analysis.
2 Flow cytometry (FCM) detection of Bcl-2, Bax gene expression: The collection of Tca8113 cells effected by different concentrations of Xanthorrhizol in 0h, 12h, 24h, 48h, 72h and control cells, according to Conventional methods of fluorescent labeling, flow cytometry fluorescence intensity to the mean fluorescence intensity channel number indicates the amount of protein expression.
Results:
1 The effect of the cell cycle of Tca8113 cells by Xanthorrhizol.
1.1 20μmol / L, 40μmol / L, 60μmol / L Xanthorrhizol, respectively, the role of Tca8113 cells, after 12h, the treatment group S + G2 / M phase cell cycle percentages compared with the control group, the difference was significant ( P <0.01); 24h drugs, each drug group S + G2 / M cell cycle phase percentage compared with the control group, the difference was significant (P <0.05); 48h drugs, each drug group S + G2 / M cell cycle phase percentage compared with the control group, the difference was significant (P <0.05); 72h drugs, each drug group S + G2 / M cell cycle phasepercentage compared with the control group, the difference was significant sex (P <0.01). The treatment group and an increase of only dimethyl sulfoxide (DMSO) did not add the solvent of drug control group, the difference was significant (P <0.05). Different time points of various drugs between groups, in addition to medication 72h later, 20μmol / L group and 40μmol / L groups showed no significant external (P> 0.05), the rest of the time the group differences were significant ( P <0.05).
1.2 The role of drug concentrations at different times of cell cycle analysis of variance showed that, 20μmol / L group, 48h and 72h between the two time points, the difference was not significant (P> 0.05), the remaining time points, the difference significant (P <0.05); 40μmol / L group, 48h and 72h the two time points, the difference was not significant (P> 0.05), the remaining time points, the difference was significant (P <0.05) ; 60μmol / L group, comparison between different time points, the differences were significant (P <0.05).
1.3 The effect of different concentrations of drugs for different time, G2 / M phase cell cycle percentage has no significant change in regularity.
2 FCM detect apoptosis of Tca8113 cells by Xanthorrhizol.
2.1 20μmol / L, 40μmol / L, 60μmol / L Xanthorrhizol on the Tca8113 cells after 12h, the apoptotic rate compared with the control group, the difference was significant (P <0.01); drugs after 24h, apoptosis rate compared with the control group, the difference was significant (P <0.01); drugs after 48h, the apoptotic rate compared with the control group, the difference was significant (P <0.01); drugs after 72h, apoptosis rate compared with the control group, the difference was significant (P <0.01); the treatment groups compare with solvent control groups which only add dimethyl sulfoxide (DMSO) did not add drug, the difference is significant (P <0.01). Different time points of various treatment groups were compared, in addition to treatment 12h after, 20μmol / L group and 40μmol / L groups showed no significant external (P> 0.05), the remaining time points of the group differences were significant (P <0.01).
2.2 The role of drug concentrations at different times of apoptosis analysis of variance showed that, in addition to 20μmol / L grouP comparison between 48h and 72h, no significant difference in external (P> 0.05), the other groups at all time points in comparison, differences were significant (P <0.05).
3 FCM detection beam acting on the Tca8113 cells apoptosis related gene protein Bcl-2, Bax expression.
3.1 Bcl-2 Protein expression:
3.1.1 12h, 24h group of proteins were compared with the control group, in addition to 20μmol / L group, other differences between the groups was significant (P <0.05); 48h group protein Compared with the control group, the difference was significant (P <0.05); 72h group protein compared with the control group, the difference was significant (P <0.05); the treatment groups compare with solvent control groups which only add dimethyl sulfoxide (DMSO) did not add drug, the difference is significant (P <0.05); at different time points between the treatment group, the difference was significant (P <0.05).
3.1.2 The drug concentrations at different time expression analysis of variance showed that, 20μmol / L group at all time points no significant difference between (P> 0.05), 40μmol / L group, in addition to 48h and 72h was no significant difference between the sex (P> 0.05), the remaining differences between each time point were significantly (P <0.05), 60μmol / L group at each time point were significant differences between (P <0.05).
3.2 Bax protein expression: The role of drug concentrations for different time, Bax protein expression in each group by factorial design analysis of variance showed that, time factors and dose factors does not exist interaction in the effection of the expression of Bax protein by Xanthorrhizol(P﹥0.05); analyzing the expression of Bax protein by drug effects in the different time and different concentration , the results show that the experimental groups and control group do not have significant differences(P﹥0.05).
Conclusion:
1 Xanthorrhizol can significantly affect the cell cycle of Tca8113 cell, which can make Cell cycle arrest in G0/G1, So as to achieve the PurPose of anti-tumor. The drug on cell cycle arrest in a time and dose dePendent.
2 Xanthorrhizol can induced aPoPtosis in Tca8113 In a certain range of concentration, the effect was time and concentration dePendently.
3 Xanthorrhizol can significantly affect the aPoPtosis-related gene Bcl-2 Protein exPression, make the Bcl-2 Protein exPression decreased Gradually, the change in Protein exPression levels in a dose-dePendent, Meanwhile, the drugs arriving to a certain Concentration,the change of Bcl-2 Protein exPression levels in time-dePendent manner.Drugs on aPoPtosis-related gene had no effect on the exPression of Bax.
方法:将复苏后的舌癌Tca8113细胞置于37℃,CO_2饱和湿度5%的常规条件下培养,待细胞进入对数生长期后用于实验。
1流式细胞术(FCM)检测细胞凋亡、细胞周期分布:收集不同浓度束骨姜黄醇作用0h、12h、24h、48h、72h的舌癌Tca8113细胞及对照组细胞,冷PBS漂洗。预冷70%乙醇4℃固定,上机检测前离心去除固定液,用0.5%胃蛋白酶消化,碘化丙锭(ProPidium Iodide,PI:50mg/L Trinton X-100 1.0%)室温避光染色。流式细胞仪上机检测,应用软件进行分析。
2流式细胞术(FCM)检测Bcl-2、Bax基因蛋白的表达情况:收集经不同浓度束骨姜黄醇作用0h、12h、24h、48h、72h的舌癌Tca8113细胞及对照组细胞,按常规方法进行荧光标记,流式细胞仪检测荧光强度,以平均荧光强度道数表示蛋白的表达量。
结果:
1束骨姜黄醇对舌癌Tca8113细胞周期的影响。
1.1 20μmol/L、40μmol/L、60μmol/L束骨姜黄醇分别作用于舌癌Tca8113细胞12h后,各用药组S+G2/M期细胞周期百分比与对照组的相比,差异具有显著性(P<0.01);药物作用24h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.05);药物作用48h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.05);药物作用72h后,各用药组S+G2/M期细胞细胞周期百分比与对照组相比,差异具有显著性(P<0.01)。各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.05)。不同时间点各用药物组间比较,除用药72h后,20μmol/L组和40μmol/L组之间差异无显著性外(P﹥0.05),其余各时间点各组比较差异均具有显著性(P<0.05)。
1.2各用药浓度组作用不同时间S+G2/M期细胞周期百分比的方差分析表明,20μmol/L组中,48h与72h两时间点之间比较,差异无显著性(P﹥0.05),其余各时间点之间,差异具有显著性(P<0.05);40μmol/L组中,48h与72h两时间点之间比较,差异无显著性(P﹥0.05),其余各时间点之间,差异具有显著性(P<0.05);60μmol/L组中,各时间点之间比较,差异均具有显著性(P<0.05)。
1.3经不同浓度药物作用不同时间后,G2/M期细胞周期百分比变化无明显规律性。
2流式细胞仪检测束骨姜黄醇对舌癌Tca8113细胞细胞凋亡的影响。
2.1 20μmol/L、40μmol/L、60μmol/L束骨姜黄醇作用于舌癌Tca8113细胞12h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用24h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用48h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);药物作用72h后,细胞凋亡率与对照组相比,差异具有显著性(P<0.01);各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.01)。不同时间点各用药组间比较,除用药12h后,20μmol/L组和40μmol/L组之间差异无显著性外(P﹥0.05),其余个时间点各组比较差异均具有显著性(P<0.01)。
2.2各用药浓度组作用不同时间细胞凋亡的方差分析表明,除20μmol/L组内48h与72h之间比较,差异无显著性外(P﹥0.05),其余各组中各时间点间比较,差异均具有显著性(P<0.05)。
3 FCM检测束骨姜黄醇作用于舌癌Tca8113细胞后凋亡相关基因蛋白Bcl-2、Bax的表达。
3.1 Bcl-2蛋白表达量:
3.1.1 12h、24h组蛋白表达分别与对照组相比,除20μmol/L组外,其余各组差异具有显著性(P<0.05);48h组蛋白表达与对照组相比,差异具有显著性(P<0.05);72h组蛋白表达与对照组相比,差异具有显著性(P<0.05);各用药组与只加二甲基亚砜(DMSO)而不加药物的各溶剂对照组相比,差异具有显著性(P<0.05);不同时间点各用药组之间比较,差异具有显著性(P<0.05)。
3.1.2各用药浓度组作用不同时间蛋白表达的方差分析表明,20μmol/L组内各时间点之间差异不显著(P﹥0.05),40μmol/L组中除48h与72h之间差异无显著性(P﹥0.05)外,其余各时间点间差异均显著(P<0.05),60μmol/L组中各时间点间差异均显著(P<0.05)。
3.2 Bax蛋白表达量:各用药浓度组作用不同时间后,各组Bax蛋白表达量经析因设计的方差分析表明,时间因素与剂量因素在束骨姜黄醇对细胞Bax蛋白表达量的影响中,不存在交互效应(P﹥0.05);逐一分析药物作用后,不同时间和不同药物浓度对Bax蛋白表达量的影响,结果表明实验组与对照组的差异不具备显著性(P﹥0.05)。
结论:
1束骨姜黄醇能够显著影响舌癌Tca8113细胞的细胞周期,使细胞周期阻滞在G0/G1期,从而达到抗肿瘤的目的。该药物对细胞周期的阻滞作用呈时间和剂量依赖关系。
2束骨姜黄醇在一定浓度范围内能够呈时间、浓度依赖性地诱导舌癌Tca8113细胞凋亡。
3束骨姜黄醇能够显著影响凋亡相关基因蛋白Bcl-2的表达,使Bcl-2蛋白表达逐渐降低,蛋白表达含量的改变呈剂量依赖性,同时该药物对Bcl-2蛋白表达的影响在达到一定浓度后出现时间依赖性;药物对凋亡相关基因蛋白Bax的表达无明显影响。
Objective: Tongue Carcinoma is the most common malignancy In the oral and maxillofacial careinoma ,in recent years, worldwide ,its incidence is increasing year by year, and its age of onset is smaller and smaller, while the degree of its malignancy is increasing. Prone to early cervical lymph node metastases, and higher transfer rate is one of the main features of tongue Carcinoma. The current treatment of choice for tongue Carcinoma is surgery, while preoperative and postoperative chemotherapy often need to meet in order to improve efficacy, but there is often a traditional chemotherapy drug resistance of tumor cells, chemotherapy drugs in patients with toxic side effects of treatment difficult to tolerate, etc. Effects of unfavorable factors, so the research and development of new drugs to treat cancer is important in a cancer therapeutic area research. Currently, scholars more and more pay attention to find some efficiency and low toxicity of the treatment of cancer drugs from plants , animals, minerals, marine and other natural substances, and has achieved considerable success. Xanthorrhizol is one of the main ingredients extracted from volatile oil of the rhizome of Curcuma xanthorrhiza,the present study shows that Xanthorrhizol can significantly inhibit the Cervical cancer, breast cancer and other malignant tumors .The subject , tongue cancer cell Line Tca8113 by in vitro ,study the Influence of Xanthorrhizol on Tca8113 cell proliferation and apoptosis, discuss the mechanism of action for tongue Carcinoma, to provide experimental basis for the chemical treatment .
Methods: After recovery, Tca8113 cells were 37℃, CO_2 saturated 5% humidity, cultured under routine conditions, until after the cells entered the logarithmic growth phase for experiments.
1 Flow cytometry (FCM) to detect apoptosis, cell cycle distribution: collect control cells and Tca8113 cells which were effected by different concentrations of Xanthorrhizol in 0h, 12h, 24h, 48h, 72h, cold PBS rinse. 4℃70% ethanol pre-cooled fixed, centrifuged to remove pre-test on the machine stationary phase, with 0.5% pepsin digestion, propidium iodide (Propidium Iodide, PI: 50mg / L Trinton X-100 1.0%) at room temperature away from light staining . Detected by flow cytometry on the machine, application software for analysis.
2 Flow cytometry (FCM) detection of Bcl-2, Bax gene expression: The collection of Tca8113 cells effected by different concentrations of Xanthorrhizol in 0h, 12h, 24h, 48h, 72h and control cells, according to Conventional methods of fluorescent labeling, flow cytometry fluorescence intensity to the mean fluorescence intensity channel number indicates the amount of protein expression.
Results:
1 The effect of the cell cycle of Tca8113 cells by Xanthorrhizol.
1.1 20μmol / L, 40μmol / L, 60μmol / L Xanthorrhizol, respectively, the role of Tca8113 cells, after 12h, the treatment group S + G2 / M phase cell cycle percentages compared with the control group, the difference was significant ( P <0.01); 24h drugs, each drug group S + G2 / M cell cycle phase percentage compared with the control group, the difference was significant (P <0.05); 48h drugs, each drug group S + G2 / M cell cycle phase percentage compared with the control group, the difference was significant (P <0.05); 72h drugs, each drug group S + G2 / M cell cycle phasepercentage compared with the control group, the difference was significant sex (P <0.01). The treatment group and an increase of only dimethyl sulfoxide (DMSO) did not add the solvent of drug control group, the difference was significant (P <0.05). Different time points of various drugs between groups, in addition to medication 72h later, 20μmol / L group and 40μmol / L groups showed no significant external (P> 0.05), the rest of the time the group differences were significant ( P <0.05).
1.2 The role of drug concentrations at different times of cell cycle analysis of variance showed that, 20μmol / L group, 48h and 72h between the two time points, the difference was not significant (P> 0.05), the remaining time points, the difference significant (P <0.05); 40μmol / L group, 48h and 72h the two time points, the difference was not significant (P> 0.05), the remaining time points, the difference was significant (P <0.05) ; 60μmol / L group, comparison between different time points, the differences were significant (P <0.05).
1.3 The effect of different concentrations of drugs for different time, G2 / M phase cell cycle percentage has no significant change in regularity.
2 FCM detect apoptosis of Tca8113 cells by Xanthorrhizol.
2.1 20μmol / L, 40μmol / L, 60μmol / L Xanthorrhizol on the Tca8113 cells after 12h, the apoptotic rate compared with the control group, the difference was significant (P <0.01); drugs after 24h, apoptosis rate compared with the control group, the difference was significant (P <0.01); drugs after 48h, the apoptotic rate compared with the control group, the difference was significant (P <0.01); drugs after 72h, apoptosis rate compared with the control group, the difference was significant (P <0.01); the treatment groups compare with solvent control groups which only add dimethyl sulfoxide (DMSO) did not add drug, the difference is significant (P <0.01). Different time points of various treatment groups were compared, in addition to treatment 12h after, 20μmol / L group and 40μmol / L groups showed no significant external (P> 0.05), the remaining time points of the group differences were significant (P <0.01).
2.2 The role of drug concentrations at different times of apoptosis analysis of variance showed that, in addition to 20μmol / L grouP comparison between 48h and 72h, no significant difference in external (P> 0.05), the other groups at all time points in comparison, differences were significant (P <0.05).
3 FCM detection beam acting on the Tca8113 cells apoptosis related gene protein Bcl-2, Bax expression.
3.1 Bcl-2 Protein expression:
3.1.1 12h, 24h group of proteins were compared with the control group, in addition to 20μmol / L group, other differences between the groups was significant (P <0.05); 48h group protein Compared with the control group, the difference was significant (P <0.05); 72h group protein compared with the control group, the difference was significant (P <0.05); the treatment groups compare with solvent control groups which only add dimethyl sulfoxide (DMSO) did not add drug, the difference is significant (P <0.05); at different time points between the treatment group, the difference was significant (P <0.05).
3.1.2 The drug concentrations at different time expression analysis of variance showed that, 20μmol / L group at all time points no significant difference between (P> 0.05), 40μmol / L group, in addition to 48h and 72h was no significant difference between the sex (P> 0.05), the remaining differences between each time point were significantly (P <0.05), 60μmol / L group at each time point were significant differences between (P <0.05).
3.2 Bax protein expression: The role of drug concentrations for different time, Bax protein expression in each group by factorial design analysis of variance showed that, time factors and dose factors does not exist interaction in the effection of the expression of Bax protein by Xanthorrhizol(P﹥0.05); analyzing the expression of Bax protein by drug effects in the different time and different concentration , the results show that the experimental groups and control group do not have significant differences(P﹥0.05).
Conclusion:
1 Xanthorrhizol can significantly affect the cell cycle of Tca8113 cell, which can make Cell cycle arrest in G0/G1, So as to achieve the PurPose of anti-tumor. The drug on cell cycle arrest in a time and dose dePendent.
2 Xanthorrhizol can induced aPoPtosis in Tca8113 In a certain range of concentration, the effect was time and concentration dePendently.
3 Xanthorrhizol can significantly affect the aPoPtosis-related gene Bcl-2 Protein exPression, make the Bcl-2 Protein exPression decreased Gradually, the change in Protein exPression levels in a dose-dePendent, Meanwhile, the drugs arriving to a certain Concentration,the change of Bcl-2 Protein exPression levels in time-dePendent manner.Drugs on aPoPtosis-related gene had no effect on the exPression of Bax.
引文
1韩晓哲,高岩.舌癌的临床及免疫病理学研究.中华口腔医学杂志,2001,36(2):112-115
2 Lebrun DP,Warnke RA.Expression of bcl-2 in fetal tissues suggests a role in morphogenesis[J].Am J Pathol,1994,142:743-753
3 Jacobson MD,Burne JF,King MP,et al.Bcl-2 blocks apoptosis in lacking mitochondrial DNA[J].Nature,1993;361:365-369
4 Olivai Z,Milliman C,Yang T,et al.Bcl-2 heterodimers in vivo with a Conserved homologen,Bax,that accelerated programmed cell death[J].Cell,1993,74:609
5蒋永和,袁继承,沈志滨.姜黄属植物化学成分的研究进展.亚太传统医药,2009,5(2):124-127
6 Lee LY, Shim JS, Rukayadi Y, et al.Antibacterial activity of xanthorrhizol isolated from Curcuma xanthorrhiza Roxb.against foodborne pathogens. J Food Prot 2008, 71:1926-1930
7 Yaya R, Dongeun Y,Jae-K H, In vitro anticandidal activity of xanthorrhizol isolated from Curcuma xanthorrhiza Roxb. Journal of Antimicrobial ChemotheraPy,2006,57:1231-1234
8 Lim CS, Jin DQ, Mok H, et al. Antioxidant and antiinflammatory activities of xanthorrhizol inhiPPocamPal neurons and primary cultured microglia. [J]Neurosci Res 2005, 82:831-838
9 Ismail N, Pihie AH, Nallapan M.Xanthorrhizol induces apoptosisvia the uP-regulation of bax and P53 in HeLa cells.AnticancerRes 2005, 25:2221-2227
10 Cheah YH, Azimahtol HL, Abdullah NR.Xanthorrhizol exhibits antiproliferative activity on MCF-7 breast cancer cells via apoptosis induction[J]. Anticancer Res 2006, 26:4527-4534
11 Seong HK, Kyoung OH, Jae KH, et al . Xanthorrhizol has a potential to attenuate the high dose cisplatin-induced nephrotoxicity in mice. Food andChemical Toxicology 2005,43:117-122
12叶志佳,舒建昌,朱海燕,等.姜黄素对人舌鳞癌Tca8113细胞增殖和凋亡作用研究[J/CD].中华口腔医学研究杂志:电子版,2009,3(3):287-291
13赵德强,李新明.姜黄素对人舌鳞癌Tca8113细胞株增殖抑制机制的研究.医药论坛杂志,2007,28(5):34-35,39
14吴大鹏,卢红,张洪志.β-榄香烯乳联合照射对人舌鳞状细胞癌裸鼠移植瘤的影响[J].郑州大学学报(医学版),2010,45(3):474-477
15吴大鹏,姜睿斌,赵德强等.β-榄香烯乳对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2):414-417
16 You JK,Kwang KP,Won YC, et al. Xanthorrhizol,a Natural Sesquiterpenoid,Induces Apoptosis and Growth Arrest in HCT116 Human Colon Cancer Cells[J]. Journal of pharmacological Sciences,2009,111:276-284
17史庆华,Adler ID,Schriever-schwemmer G,等.秋水仙碱(COM)诱发小鼠骨髓细胞C-有丝分裂效应的研究.细胞生物学杂志,1996,18(4):172-176
18 Demarcq C,Bunch RT,Creswell D,et al.The role of cycle progression in cisPlatin-induced apoptosis in chinese hamster ovary cells [J] .Cell Growth Differ.1994,5(9):983-993
19 Majong JI. Apoptosis oncosis and necrosis: an overview of cell death[J]. Am J Pathol, 1995,146(1):3-15
20 Adams JM, Cory S: The Bcl-2 Protein family: Arbiters of cell survival[J]. Science 281:1309-1312, 1998
21 Strasser A, Huang DCS, Vaux DL: The role of the bcl-2/ced-9 gene family in cancer and the general implications in cell death control for tumorigenesis and resistance to chemotheraPy[J]. Biochim Biophys Acta 1997,1333:151-178
22 Scott N,Hale A,Deakin M,et al.A histopathological assessment of the response of rectal adenocarcinoma to combination chemo-radiotherapy:relationship to apoptotic activity, p53 and bcl-2 expression[J].Eur J Surg Oncol,1998,24:169-173
23刘善廷,魏林,秦建武,等.舌鳞癌中bcl-2的表达及意义.医学论坛杂志,2009, 30(3):11-12
24郅克谦,袁常莉,林兆全,等.舌癌术前化疗PCNA、bcl-2、Bax基因表达及临床意义.新疆医科大学学报,2001,11,24(4):328-330
25 Metenssian S,Kontogiannea M.Fas antigen expression and function in human colorectal carcinoma.correlation with Bcl-2 expression[J].Proc Am Asso Cancer Res,1996,37:15
26彭玮丹,张杰,惠宏襄,等.bcl-2核酶与bax在诱导食管癌细胞凋亡中的协同作用[J].中国生物化学与分子生物学学报[J],2000,16(6):827-831
27 Tri H,Sharifah S, Meenakshii N,et al. Regulation of P53-,Bcl-2- and CasPase-dePendent Signaling Pathway in Xanthorrhizol-induced APoPtosis of HePG2 HePatoma Cells. Anticancer Res,2007,27: 965-971
28王彤,刘存志,刘玉珍等. bcl-2/bax基因调控机体细胞凋亡的机制研究进展[J].中国老年学杂志,2008,28(16):1658-1660
1蒋永和,袁继承,沈志滨.姜黄属植物化学成分的研究进展[J].亚太传统医药,2009,5(2):124-127
2王琰,王慕邹.姜黄属常用中药的研究进展[J].中国药学杂志,2001,36(2):80-84
3舒建昌,叶国荣,吕霞,等.姜黄素治疗肝纤维化及其作用机制的初步研究.中华肝脏病杂志,2007,15(10):753-757
4赵心宇,孟秀香,贾莉,等.姜黄素抗肿瘤机制.实用药物与临床,2006,9(1):51-52
5许东晖,王胜,金晶,等.姜黄素的药理作用研究进展[J].中草药,2005,36(11):1737-1740
6 Bush JA,Cheung KJ Jr,Li G. Curcumin induces apoptosis in human melanoma cells through a Fas receptor/caspase-8 pathway independent of p53. ExP Cell Res,2001,271(2):305-314
7 Shishodia S,Amin HM,Lai R,et al.Curcumin (diferuloyl-methane) inhibits constitutive NF-kappa B activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma[J].Biochem Pharmaco,2005, 70 (4):700-713
8 Rajasingh J, Raikwar HP,Muthian G, et al.Curcumin induces growth-arrest and apoptosis in association with the inhibition of constitutively active JAK-STAT pathway in T cell leukemia [J].Biochem Biophys Res Commun, 2006, 340(2):359-368
9 Liu HL,Chen Y,Cui GH,et al. Curcumin,a potent anti-tumor reagent, is a novel histone deacetylase inhibit or regulating B-NHL cell line Raji proliferation[J]. Acta Pharmacol Sin,2005,26(5):603-609
10 Aggarwal BB, Shishodia S,Takada Y, et al.Curcumin suppresses the paclitaxel-induced nuclear factor-kappa B pathway in breast cancer cells and inhibits lung metastasis of human breast cancer in nude mice[J].Clin CancerRes, 2005, 11(20): 7490-7498
11周钱梅,苏式兵,张辉,等.姜黄素对乳腺癌MCF-7细胞信号传导通路相关蛋白激酶的调节作用[J].中药材, 2009, 32(5): 728-732
12黄冬生,李军.姜黄素对人肺癌细胞凋亡相关蛋白Survivin表达的影响[J].临床肺科杂志, 2009, 14(6): 723-724
13 Duvoix A, Morceau F, Schnekenburger M, et al. rcumin-induced cell death in two leukemia cell lines:K562 and Jurkat[J]. Ann NY Acad Sci,2006,1010:389-392
14叶志佳,舒建昌,朱海燕,等.姜黄素对人舌鳞癌Tca8113细胞增殖和凋亡作用研究[J/CD].中华口腔医学研究杂志:电子版,2009,( 3):287-291
15赵德强,李新明.姜黄素对人舌鳞癌Tca8113细胞株增殖抑制机制的研究.医药论坛杂志,2007,28(5):34-35,39
16 BanerjiA,ChakrabartiJ,MitraA,et al. Effect of curcumin on gelatinase A(MMP-2) activity in B16F10 melanoma cells.Cancer Lett,2004,211(2):235-242
17 Kunnumakkara AB , Anand P , Aggarwal BB.Curcumin inhibits proliferation,invasion,angiogenesis and metastasis of different cancers through interaction with multiPle cell signaling proteins.Cancer Lett,2008;269:199-255
18 Narasimhan M,Ammanamanchi S. Curcumin blocks RON tyrosine kinase-mediated invasion of breast carcinoma cells.Cancer Res,2008,68:5185-5192
19 Weir NV,Selvendiran K,Kutala VK,et al. Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK.Cancer Biol Ther ,2007,6:178-184
20 Aravindan N,Madhusoodhanan R,Ahmad S,Johnson D,Herman TS. Curcumin inhibits NFkappaB mediated radioprotection and modulate apoptosis related genes in human neuroblastoma cells. Cancer Biol Ther,2008,7:569-576
21 umar A,Dhawan S,Hardegen NJ,et al.Curcumin(diferuloylmethane) inhibition of tumor necrosis factor(TNF)-mediated adhesion of monocytes to endothelialcells by suppression of cell surface expression of adhesionmolecules and of nuclear factor-kappaB activation[J].Bio-chem Pharmacol,1998,55(6):775-783
22汪茜,汪宏波.姜黄素对IGF-1调控的宫颈癌淋巴转移的影响[J].数理医药学杂志,2009,22(3):290-292
23 nuchaPreeda S,Leechanachai P,Smith MM,et al. Modulation of P-glycoProtein expression and function by curcumin in multidrug-resistant human KB cells[J].Biochem Pharmacol,2002,64(4):573-582
24 Chan MM,Fong D,SoPrano KJ,et al. Inhibition of growth and sensitization to cisplation-mediated killing of ovarian cancer cells by polyphenolic chemopreventive agents[J].J Cell Physiol , 2003 ,194(1):63-70
25李广朝.β-榄香烯的基础研究与临床应用[J].长春中医药大学学报, 2009(2): 185-186
26陆羡,向金峰.β-榄香烯对K562细胞周期与细胞凋亡的影响及其机制探讨[J].中国小儿血液与肿瘤杂志,2008,13(4):149-152
27方卓,陈洪胜,陈宏勃,等.榄香烯对PLA801D细胞株诱导分化研究[J].中国老年杂志, 2008, 28(8): 768-769
28 Wang G,Li X,Huang F,et al.Antitumor effect ofβ-elemene in non-small-cell lung cancer cells is mediated via induction of cell cycle arrest and apoptotic cell death[J].Cell Mol Life Sci,2005,62:881–893
29刘跃明,孙立群,王义.榄香烯诱导HeP-2细胞凋亡及其对细胞周期各时相的影响[J].中国老年学杂志,2008,28(9):868-869
30 Li X,Wang G,Zhao J,et al.Antiproliferative effect ofβ-elemene in chemoresistant ovarian carcinoma cells is mediated through arrest of the cell cycle at the G2-M phase[J].Cell Mol Life Sci,2005,62:894–904
31陈春美,杨卫忠,王春华,等.榄香烯诱导人脑胶质瘤U251细胞凋亡及对端粒酶活性和hTERT表达影响的研究[J].中国肿瘤临床,2006,35(5):280-283
32 Jiang Hao,Ma Shenglin,Feng Jianguo.In vitro study of radiosensitization byβ-Elemene in A549 cell line from adenocarcinoma of lung[J]. Chin Ger J Clin Oncol,2009,8(1):12-15
33 Zhao J,Li Q Q,Zou B,et al.In vitro combination characterization of the new anticancer Plant drug beta-elemene with taxanes against human lung carcinoma[J].Int J oncol,2007,31(2):241-252
34李传刚,李墨林,周琴,等.β-榄香烯对人膀胱癌BIU-87细胞磷脂膜功能及Bcl-2表达的影响[J].中草药,2007,36(6):886-889
35 Hu J,Jin W,Yang PM.Reversal of resistance to adriamycin in human breast cancer cell line MCF-7/ADM by beta-elemene[J].Chin J Oncol,2004,26(5):268-270
36陈浩,师亮,成志勇,姚丽,杨营营,潘崚.β-榄香烯对人骨髓瘤细胞RPM I-8226增殖与凋亡的影响[J].中国实验血液学杂志.2010,18(2):368-371
37魏为,唐罗生,李立,等.榄香烯乳联合足叶乙苷对视网膜母细胞瘤HXO-RB44细胞凋亡及survivin表达的影响[J].国际眼科杂志,2007,7(1):64-66
38 She JJ,Wang ZM,Che XM,et al.Research of beta-elemene interventional treatment on VX2 carcinoma transplanted on kidney in rabbits[J].J Chin integ Med,2006,4(6):611-614
39 Mukherjee S,Ghosh U,Bhattacharyya N P,et al. Curcumin-induced apoptosis in human leukemia cell HL-60 is associated with inhibition of telomerase activity[J].Mol Cell Biochem,2007,29(7):31-39
40 Wang Yi,Fang Mei-Yun. Effect of Ginseng saPonin,arse trioxide,β-elemene combined with CTX on Telomere-Telomerase system in K562 cell line[J].J Exp Hematol,2006,14(6):1089-1095
41周昆,崔黎,闫焱,等.榄香烯对人肺腺癌SPC-A-1细胞VEGF-C及VEGFR-3表达的影响[J].中国老年学杂志,2008,28(5):551-553
42 Cao Z,Ji W.Elemene in the treatment of the transplantable model of human laryngeal squamous carcinoma in nude mice[J].Chin J otorhinolaryngol head and neck surg,2007,21(9):417-420
43郑瑾,刘强,任凯夕,等.β-榄香烯对肝癌细胞SK-hep-1的迁移和侵袭力的影响[J].现代肿瘤医学, 2009, 17(11):2054-2058
44 Tan G,Wang ZY,Che LQ,et al. Immunotherapeutic effects on murine pancreatic carcinoma byβ-elemene combined with dendritic cells modified with genes encoding interleukin-23[J].Front Med Chin,2007,1(1):41-45
45衣服新,黄强,周丽英.温莪术提取物协同抗CD3-抗胶质瘤双特异性抗体抑制神经胶质瘤生长[J].中国临床复,2006,10(23):124-126
46姚淑娟,刘伯阳,吕丽艳.榄香烯提高肿瘤化疗药物疗效及抗免疫抑制作用的研究[J].中医药学刊, 2006, 24 (3): 456-457
47官成浓,何国章,银正民.榄香烯乳配合化疗对进展期胃癌患者免疫功能的影响[J].中国肿瘤临床, 2001, 28 (2): 123-124
48大鹏,卢红,张洪志.β-榄香烯乳联合照射对人舌鳞状细胞癌裸鼠移植瘤的影响[J].郑州大学学报(医学版),2010,45(3):474-477
49 Jiang H,Ma SL,Feng JG. In vitro study of radiosensitization byβ-Elemene inA549 cell line from adenocarcinoma of lung[J].Chi-nese-German Journal ofClinicalOncology, 2009, 8(1): 12-15
50陈春美,杨卫忠,王春华,等.榄香烯对人脑胶质瘤U251 /ADM耐药细胞株多药耐药性的逆转作用[J].中华实验外科杂志,2006, 23(4): 601-603
51郝立宏,赵瑾瑶,高船舟,等.β-榄香烯对K562/阿霉素细胞多药耐药性的逆转及其对P-糖蛋白表达的影响[J].解剖学报,2006,37(1):48-51
52高海德,张佩禹,赵荣宇,等.榄香烯对小鼠瘤株Hca-A2/p化疗的影响[J].中华肿瘤防治杂志,2006,13(7):492-494
53郝立宏,卢步峰,于丽敏,等.β-榄香烯吗素与阿霉素联合应用对CEM /ADM细胞生长的影响[J].大连医科大学学报,2002, 22(3): 165-167
54向彬,张福胤,吴佩春.β-榄香烯及顺铂对人涎腺腺样囊性癌SACC-83细胞毒作用的研究[J].现代口腔医学杂志.2005,19(4):433-434
55徐莉英,王宪明,于志瀛,董金华.β-榄香烯胺类衍生物的合成及抗癌活性研究[J].中国药物化学杂志。2009,19(4):247-250、260
56于志瀛.β-榄香烯衍生物(ET)诱导NB4细胞凋亡的机制研究[J].中国中药杂志,2010,35(10):1324-1327
57陈光,丁晓飞,肖晋芳,等.榄香烯衍生物诱导HeLa细胞凋亡的实验研究[J].中国药理学通报,2007,23(2):246-250
58徐立春,边可君,刘志敏,等.天然药物莪术醇抑制肿瘤细胞生长及RNA合成影响的初步研究[J].肿瘤, 2005, 25(6): 570-572
59徐立春,卜平,许祥裕,等.中药莪术醇构建胃癌细胞疫苗对胃癌的实验治疗效果[J].中医杂志, 2007 48(5):450-452
60徐立春,边可君,卜平,等.莪术醇修饰的肿瘤疫苗抗MFC效应的研究[J].现代肿瘤医学,2006,14(11):1357-1359
61徐立春,陈平,卜平,等.莪术醇自体瘤苗治疗胃癌的初步研究[J].中国肿瘤防治杂志,2009,16(20):1587-1589
62唐渊,李晓辉.莪术提取物对肝癌细胞系HePG2的抗癌作用及机制研究[J].中国药理学通报,2007,23(6):790-794
63杜小燕,丰培勋,侯颖,覃华,韩艳,张琰.莪术醇抑制人结肠癌SW1116细胞增殖及相关机制的研究[J].科学技术与工程,2010,10(28):6967-6970
64林海,李晓辉.莪术醇诱导白血病L1210细胞凋亡作用研究[J].中国药房, 2008, 19(30): 2328-2329
65林海,李晓辉.莪术醇诱导慢性粒细胞白血病K562细胞分化的研究[J].现代生物学进展, 2007, 7(11): 1674-1676
66 Ismail N, Pihie AH, NallaPan M: Xanthorrhizol induces apoptosisvia the up-regulation of bax and p53 in HeLa cells.AnticancerRes 2005, 25:2221-2227
67 Cheah YH, Azimahtol HL, Abdullah NR.Xanthorrhizol exhibits antiProliferative activity on MCF-7 breast cancer cells via apoptosis induction. Anticancer Res 2006, 26:4527-4534
68 Choi MA , Kim Sh, Chung WY, et al.Xanthorrhizol , a natural sesquiterpenoid from Curcuma xanthorrhiza, has an anti-metastatic potential in experimental mouse lung metastatic model. Biochem Biophys Res Comm 2005, 326:210-217
69 Kim SH, Hong KO, Chung WY, et al. Abbrogation of cisplatin-induced hepatotoxicity in mice by xanthorrhizol is related to its effects on the regulation of gene transcription.Toxicol Appl Pharmacol ,2004, 196:346-355
70 Seong HK ,Kyoung OH, Jae KH, et al. Xanthorrhizol has a potential to attenuate the high dose cisplatin-induced nephrotoxicity in mice[J]. Food and Chemical Toxicology,2005,43:117-122
2 Lebrun DP,Warnke RA.Expression of bcl-2 in fetal tissues suggests a role in morphogenesis[J].Am J Pathol,1994,142:743-753
3 Jacobson MD,Burne JF,King MP,et al.Bcl-2 blocks apoptosis in lacking mitochondrial DNA[J].Nature,1993;361:365-369
4 Olivai Z,Milliman C,Yang T,et al.Bcl-2 heterodimers in vivo with a Conserved homologen,Bax,that accelerated programmed cell death[J].Cell,1993,74:609
5蒋永和,袁继承,沈志滨.姜黄属植物化学成分的研究进展.亚太传统医药,2009,5(2):124-127
6 Lee LY, Shim JS, Rukayadi Y, et al.Antibacterial activity of xanthorrhizol isolated from Curcuma xanthorrhiza Roxb.against foodborne pathogens. J Food Prot 2008, 71:1926-1930
7 Yaya R, Dongeun Y,Jae-K H, In vitro anticandidal activity of xanthorrhizol isolated from Curcuma xanthorrhiza Roxb. Journal of Antimicrobial ChemotheraPy,2006,57:1231-1234
8 Lim CS, Jin DQ, Mok H, et al. Antioxidant and antiinflammatory activities of xanthorrhizol inhiPPocamPal neurons and primary cultured microglia. [J]Neurosci Res 2005, 82:831-838
9 Ismail N, Pihie AH, Nallapan M.Xanthorrhizol induces apoptosisvia the uP-regulation of bax and P53 in HeLa cells.AnticancerRes 2005, 25:2221-2227
10 Cheah YH, Azimahtol HL, Abdullah NR.Xanthorrhizol exhibits antiproliferative activity on MCF-7 breast cancer cells via apoptosis induction[J]. Anticancer Res 2006, 26:4527-4534
11 Seong HK, Kyoung OH, Jae KH, et al . Xanthorrhizol has a potential to attenuate the high dose cisplatin-induced nephrotoxicity in mice. Food andChemical Toxicology 2005,43:117-122
12叶志佳,舒建昌,朱海燕,等.姜黄素对人舌鳞癌Tca8113细胞增殖和凋亡作用研究[J/CD].中华口腔医学研究杂志:电子版,2009,3(3):287-291
13赵德强,李新明.姜黄素对人舌鳞癌Tca8113细胞株增殖抑制机制的研究.医药论坛杂志,2007,28(5):34-35,39
14吴大鹏,卢红,张洪志.β-榄香烯乳联合照射对人舌鳞状细胞癌裸鼠移植瘤的影响[J].郑州大学学报(医学版),2010,45(3):474-477
15吴大鹏,姜睿斌,赵德强等.β-榄香烯乳对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2):414-417
16 You JK,Kwang KP,Won YC, et al. Xanthorrhizol,a Natural Sesquiterpenoid,Induces Apoptosis and Growth Arrest in HCT116 Human Colon Cancer Cells[J]. Journal of pharmacological Sciences,2009,111:276-284
17史庆华,Adler ID,Schriever-schwemmer G,等.秋水仙碱(COM)诱发小鼠骨髓细胞C-有丝分裂效应的研究.细胞生物学杂志,1996,18(4):172-176
18 Demarcq C,Bunch RT,Creswell D,et al.The role of cycle progression in cisPlatin-induced apoptosis in chinese hamster ovary cells [J] .Cell Growth Differ.1994,5(9):983-993
19 Majong JI. Apoptosis oncosis and necrosis: an overview of cell death[J]. Am J Pathol, 1995,146(1):3-15
20 Adams JM, Cory S: The Bcl-2 Protein family: Arbiters of cell survival[J]. Science 281:1309-1312, 1998
21 Strasser A, Huang DCS, Vaux DL: The role of the bcl-2/ced-9 gene family in cancer and the general implications in cell death control for tumorigenesis and resistance to chemotheraPy[J]. Biochim Biophys Acta 1997,1333:151-178
22 Scott N,Hale A,Deakin M,et al.A histopathological assessment of the response of rectal adenocarcinoma to combination chemo-radiotherapy:relationship to apoptotic activity, p53 and bcl-2 expression[J].Eur J Surg Oncol,1998,24:169-173
23刘善廷,魏林,秦建武,等.舌鳞癌中bcl-2的表达及意义.医学论坛杂志,2009, 30(3):11-12
24郅克谦,袁常莉,林兆全,等.舌癌术前化疗PCNA、bcl-2、Bax基因表达及临床意义.新疆医科大学学报,2001,11,24(4):328-330
25 Metenssian S,Kontogiannea M.Fas antigen expression and function in human colorectal carcinoma.correlation with Bcl-2 expression[J].Proc Am Asso Cancer Res,1996,37:15
26彭玮丹,张杰,惠宏襄,等.bcl-2核酶与bax在诱导食管癌细胞凋亡中的协同作用[J].中国生物化学与分子生物学学报[J],2000,16(6):827-831
27 Tri H,Sharifah S, Meenakshii N,et al. Regulation of P53-,Bcl-2- and CasPase-dePendent Signaling Pathway in Xanthorrhizol-induced APoPtosis of HePG2 HePatoma Cells. Anticancer Res,2007,27: 965-971
28王彤,刘存志,刘玉珍等. bcl-2/bax基因调控机体细胞凋亡的机制研究进展[J].中国老年学杂志,2008,28(16):1658-1660
1蒋永和,袁继承,沈志滨.姜黄属植物化学成分的研究进展[J].亚太传统医药,2009,5(2):124-127
2王琰,王慕邹.姜黄属常用中药的研究进展[J].中国药学杂志,2001,36(2):80-84
3舒建昌,叶国荣,吕霞,等.姜黄素治疗肝纤维化及其作用机制的初步研究.中华肝脏病杂志,2007,15(10):753-757
4赵心宇,孟秀香,贾莉,等.姜黄素抗肿瘤机制.实用药物与临床,2006,9(1):51-52
5许东晖,王胜,金晶,等.姜黄素的药理作用研究进展[J].中草药,2005,36(11):1737-1740
6 Bush JA,Cheung KJ Jr,Li G. Curcumin induces apoptosis in human melanoma cells through a Fas receptor/caspase-8 pathway independent of p53. ExP Cell Res,2001,271(2):305-314
7 Shishodia S,Amin HM,Lai R,et al.Curcumin (diferuloyl-methane) inhibits constitutive NF-kappa B activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma[J].Biochem Pharmaco,2005, 70 (4):700-713
8 Rajasingh J, Raikwar HP,Muthian G, et al.Curcumin induces growth-arrest and apoptosis in association with the inhibition of constitutively active JAK-STAT pathway in T cell leukemia [J].Biochem Biophys Res Commun, 2006, 340(2):359-368
9 Liu HL,Chen Y,Cui GH,et al. Curcumin,a potent anti-tumor reagent, is a novel histone deacetylase inhibit or regulating B-NHL cell line Raji proliferation[J]. Acta Pharmacol Sin,2005,26(5):603-609
10 Aggarwal BB, Shishodia S,Takada Y, et al.Curcumin suppresses the paclitaxel-induced nuclear factor-kappa B pathway in breast cancer cells and inhibits lung metastasis of human breast cancer in nude mice[J].Clin CancerRes, 2005, 11(20): 7490-7498
11周钱梅,苏式兵,张辉,等.姜黄素对乳腺癌MCF-7细胞信号传导通路相关蛋白激酶的调节作用[J].中药材, 2009, 32(5): 728-732
12黄冬生,李军.姜黄素对人肺癌细胞凋亡相关蛋白Survivin表达的影响[J].临床肺科杂志, 2009, 14(6): 723-724
13 Duvoix A, Morceau F, Schnekenburger M, et al. rcumin-induced cell death in two leukemia cell lines:K562 and Jurkat[J]. Ann NY Acad Sci,2006,1010:389-392
14叶志佳,舒建昌,朱海燕,等.姜黄素对人舌鳞癌Tca8113细胞增殖和凋亡作用研究[J/CD].中华口腔医学研究杂志:电子版,2009,( 3):287-291
15赵德强,李新明.姜黄素对人舌鳞癌Tca8113细胞株增殖抑制机制的研究.医药论坛杂志,2007,28(5):34-35,39
16 BanerjiA,ChakrabartiJ,MitraA,et al. Effect of curcumin on gelatinase A(MMP-2) activity in B16F10 melanoma cells.Cancer Lett,2004,211(2):235-242
17 Kunnumakkara AB , Anand P , Aggarwal BB.Curcumin inhibits proliferation,invasion,angiogenesis and metastasis of different cancers through interaction with multiPle cell signaling proteins.Cancer Lett,2008;269:199-255
18 Narasimhan M,Ammanamanchi S. Curcumin blocks RON tyrosine kinase-mediated invasion of breast carcinoma cells.Cancer Res,2008,68:5185-5192
19 Weir NV,Selvendiran K,Kutala VK,et al. Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK.Cancer Biol Ther ,2007,6:178-184
20 Aravindan N,Madhusoodhanan R,Ahmad S,Johnson D,Herman TS. Curcumin inhibits NFkappaB mediated radioprotection and modulate apoptosis related genes in human neuroblastoma cells. Cancer Biol Ther,2008,7:569-576
21 umar A,Dhawan S,Hardegen NJ,et al.Curcumin(diferuloylmethane) inhibition of tumor necrosis factor(TNF)-mediated adhesion of monocytes to endothelialcells by suppression of cell surface expression of adhesionmolecules and of nuclear factor-kappaB activation[J].Bio-chem Pharmacol,1998,55(6):775-783
22汪茜,汪宏波.姜黄素对IGF-1调控的宫颈癌淋巴转移的影响[J].数理医药学杂志,2009,22(3):290-292
23 nuchaPreeda S,Leechanachai P,Smith MM,et al. Modulation of P-glycoProtein expression and function by curcumin in multidrug-resistant human KB cells[J].Biochem Pharmacol,2002,64(4):573-582
24 Chan MM,Fong D,SoPrano KJ,et al. Inhibition of growth and sensitization to cisplation-mediated killing of ovarian cancer cells by polyphenolic chemopreventive agents[J].J Cell Physiol , 2003 ,194(1):63-70
25李广朝.β-榄香烯的基础研究与临床应用[J].长春中医药大学学报, 2009(2): 185-186
26陆羡,向金峰.β-榄香烯对K562细胞周期与细胞凋亡的影响及其机制探讨[J].中国小儿血液与肿瘤杂志,2008,13(4):149-152
27方卓,陈洪胜,陈宏勃,等.榄香烯对PLA801D细胞株诱导分化研究[J].中国老年杂志, 2008, 28(8): 768-769
28 Wang G,Li X,Huang F,et al.Antitumor effect ofβ-elemene in non-small-cell lung cancer cells is mediated via induction of cell cycle arrest and apoptotic cell death[J].Cell Mol Life Sci,2005,62:881–893
29刘跃明,孙立群,王义.榄香烯诱导HeP-2细胞凋亡及其对细胞周期各时相的影响[J].中国老年学杂志,2008,28(9):868-869
30 Li X,Wang G,Zhao J,et al.Antiproliferative effect ofβ-elemene in chemoresistant ovarian carcinoma cells is mediated through arrest of the cell cycle at the G2-M phase[J].Cell Mol Life Sci,2005,62:894–904
31陈春美,杨卫忠,王春华,等.榄香烯诱导人脑胶质瘤U251细胞凋亡及对端粒酶活性和hTERT表达影响的研究[J].中国肿瘤临床,2006,35(5):280-283
32 Jiang Hao,Ma Shenglin,Feng Jianguo.In vitro study of radiosensitization byβ-Elemene in A549 cell line from adenocarcinoma of lung[J]. Chin Ger J Clin Oncol,2009,8(1):12-15
33 Zhao J,Li Q Q,Zou B,et al.In vitro combination characterization of the new anticancer Plant drug beta-elemene with taxanes against human lung carcinoma[J].Int J oncol,2007,31(2):241-252
34李传刚,李墨林,周琴,等.β-榄香烯对人膀胱癌BIU-87细胞磷脂膜功能及Bcl-2表达的影响[J].中草药,2007,36(6):886-889
35 Hu J,Jin W,Yang PM.Reversal of resistance to adriamycin in human breast cancer cell line MCF-7/ADM by beta-elemene[J].Chin J Oncol,2004,26(5):268-270
36陈浩,师亮,成志勇,姚丽,杨营营,潘崚.β-榄香烯对人骨髓瘤细胞RPM I-8226增殖与凋亡的影响[J].中国实验血液学杂志.2010,18(2):368-371
37魏为,唐罗生,李立,等.榄香烯乳联合足叶乙苷对视网膜母细胞瘤HXO-RB44细胞凋亡及survivin表达的影响[J].国际眼科杂志,2007,7(1):64-66
38 She JJ,Wang ZM,Che XM,et al.Research of beta-elemene interventional treatment on VX2 carcinoma transplanted on kidney in rabbits[J].J Chin integ Med,2006,4(6):611-614
39 Mukherjee S,Ghosh U,Bhattacharyya N P,et al. Curcumin-induced apoptosis in human leukemia cell HL-60 is associated with inhibition of telomerase activity[J].Mol Cell Biochem,2007,29(7):31-39
40 Wang Yi,Fang Mei-Yun. Effect of Ginseng saPonin,arse trioxide,β-elemene combined with CTX on Telomere-Telomerase system in K562 cell line[J].J Exp Hematol,2006,14(6):1089-1095
41周昆,崔黎,闫焱,等.榄香烯对人肺腺癌SPC-A-1细胞VEGF-C及VEGFR-3表达的影响[J].中国老年学杂志,2008,28(5):551-553
42 Cao Z,Ji W.Elemene in the treatment of the transplantable model of human laryngeal squamous carcinoma in nude mice[J].Chin J otorhinolaryngol head and neck surg,2007,21(9):417-420
43郑瑾,刘强,任凯夕,等.β-榄香烯对肝癌细胞SK-hep-1的迁移和侵袭力的影响[J].现代肿瘤医学, 2009, 17(11):2054-2058
44 Tan G,Wang ZY,Che LQ,et al. Immunotherapeutic effects on murine pancreatic carcinoma byβ-elemene combined with dendritic cells modified with genes encoding interleukin-23[J].Front Med Chin,2007,1(1):41-45
45衣服新,黄强,周丽英.温莪术提取物协同抗CD3-抗胶质瘤双特异性抗体抑制神经胶质瘤生长[J].中国临床复,2006,10(23):124-126
46姚淑娟,刘伯阳,吕丽艳.榄香烯提高肿瘤化疗药物疗效及抗免疫抑制作用的研究[J].中医药学刊, 2006, 24 (3): 456-457
47官成浓,何国章,银正民.榄香烯乳配合化疗对进展期胃癌患者免疫功能的影响[J].中国肿瘤临床, 2001, 28 (2): 123-124
48大鹏,卢红,张洪志.β-榄香烯乳联合照射对人舌鳞状细胞癌裸鼠移植瘤的影响[J].郑州大学学报(医学版),2010,45(3):474-477
49 Jiang H,Ma SL,Feng JG. In vitro study of radiosensitization byβ-Elemene inA549 cell line from adenocarcinoma of lung[J].Chi-nese-German Journal ofClinicalOncology, 2009, 8(1): 12-15
50陈春美,杨卫忠,王春华,等.榄香烯对人脑胶质瘤U251 /ADM耐药细胞株多药耐药性的逆转作用[J].中华实验外科杂志,2006, 23(4): 601-603
51郝立宏,赵瑾瑶,高船舟,等.β-榄香烯对K562/阿霉素细胞多药耐药性的逆转及其对P-糖蛋白表达的影响[J].解剖学报,2006,37(1):48-51
52高海德,张佩禹,赵荣宇,等.榄香烯对小鼠瘤株Hca-A2/p化疗的影响[J].中华肿瘤防治杂志,2006,13(7):492-494
53郝立宏,卢步峰,于丽敏,等.β-榄香烯吗素与阿霉素联合应用对CEM /ADM细胞生长的影响[J].大连医科大学学报,2002, 22(3): 165-167
54向彬,张福胤,吴佩春.β-榄香烯及顺铂对人涎腺腺样囊性癌SACC-83细胞毒作用的研究[J].现代口腔医学杂志.2005,19(4):433-434
55徐莉英,王宪明,于志瀛,董金华.β-榄香烯胺类衍生物的合成及抗癌活性研究[J].中国药物化学杂志。2009,19(4):247-250、260
56于志瀛.β-榄香烯衍生物(ET)诱导NB4细胞凋亡的机制研究[J].中国中药杂志,2010,35(10):1324-1327
57陈光,丁晓飞,肖晋芳,等.榄香烯衍生物诱导HeLa细胞凋亡的实验研究[J].中国药理学通报,2007,23(2):246-250
58徐立春,边可君,刘志敏,等.天然药物莪术醇抑制肿瘤细胞生长及RNA合成影响的初步研究[J].肿瘤, 2005, 25(6): 570-572
59徐立春,卜平,许祥裕,等.中药莪术醇构建胃癌细胞疫苗对胃癌的实验治疗效果[J].中医杂志, 2007 48(5):450-452
60徐立春,边可君,卜平,等.莪术醇修饰的肿瘤疫苗抗MFC效应的研究[J].现代肿瘤医学,2006,14(11):1357-1359
61徐立春,陈平,卜平,等.莪术醇自体瘤苗治疗胃癌的初步研究[J].中国肿瘤防治杂志,2009,16(20):1587-1589
62唐渊,李晓辉.莪术提取物对肝癌细胞系HePG2的抗癌作用及机制研究[J].中国药理学通报,2007,23(6):790-794
63杜小燕,丰培勋,侯颖,覃华,韩艳,张琰.莪术醇抑制人结肠癌SW1116细胞增殖及相关机制的研究[J].科学技术与工程,2010,10(28):6967-6970
64林海,李晓辉.莪术醇诱导白血病L1210细胞凋亡作用研究[J].中国药房, 2008, 19(30): 2328-2329
65林海,李晓辉.莪术醇诱导慢性粒细胞白血病K562细胞分化的研究[J].现代生物学进展, 2007, 7(11): 1674-1676
66 Ismail N, Pihie AH, NallaPan M: Xanthorrhizol induces apoptosisvia the up-regulation of bax and p53 in HeLa cells.AnticancerRes 2005, 25:2221-2227
67 Cheah YH, Azimahtol HL, Abdullah NR.Xanthorrhizol exhibits antiProliferative activity on MCF-7 breast cancer cells via apoptosis induction. Anticancer Res 2006, 26:4527-4534
68 Choi MA , Kim Sh, Chung WY, et al.Xanthorrhizol , a natural sesquiterpenoid from Curcuma xanthorrhiza, has an anti-metastatic potential in experimental mouse lung metastatic model. Biochem Biophys Res Comm 2005, 326:210-217
69 Kim SH, Hong KO, Chung WY, et al. Abbrogation of cisplatin-induced hepatotoxicity in mice by xanthorrhizol is related to its effects on the regulation of gene transcription.Toxicol Appl Pharmacol ,2004, 196:346-355
70 Seong HK ,Kyoung OH, Jae KH, et al. Xanthorrhizol has a potential to attenuate the high dose cisplatin-induced nephrotoxicity in mice[J]. Food and Chemical Toxicology,2005,43:117-122