胃复方含药血浆对人胃癌BGC-823细胞增殖影响的实验研究
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摘要
目的基于过氧化物酶体增殖体激活受体γ(PPARγ)、垂体肿瘤转化基因(PTTG)途径探讨胃复方含药血浆对人胃癌BGC-823细胞株增殖的影响。方法制备胃复方含药血浆,用不同浓度含药血浆作用人胃癌BGC-823细胞株,24 h、48 h、72 h后分别用MTS法检测细胞增殖抑制率并计算半数抑制率IC_(50)。并以1/4IC_(50)浓度胃复方含药血浆作用胃癌BGC-823细胞株72 h后,流式细胞术检测细胞凋亡变化、Western blot方法测定人胃癌BGC-823细胞中PPARγ、PTTG、原癌基因(c-Myc)、端粒酶亚单位(h TERT)蛋白表达情况。结果胃复方含药血浆对人胃癌BGC-823细胞有明显的增殖抑制作用,且呈剂量及时间依赖性,5%、10%、20%浓度的药物干预后各组间比较差异有统计学意义(P <0. 05),而10%浓度以上的各药物组细胞生长大部分停滞,24 h、48 h各组间比较差异无统计学意义(P> 0. 05),72 h各组间比较差异有统计学意义(P <0. 05)。胃复方含药血浆低、中、高剂量组IC_(50)分别为30. 32%、19. 37%、14. 69%。与空白组、PBS阴性对照组凋亡率比较,胃复方组凋亡率升高(P <0. 01)。与PBS阴性对照组比较,中剂量1/4IC_(50)含药血浆作用细胞72 h后,PPARγ蛋白表达升高(P <0. 05,P <0. 01),PTTG、c-Myc、hTERT蛋白表达下调(P <0. 01)。结论胃复方含药血浆对人胃癌BGC-823细胞有增殖抑制作用,呈时间和浓度依赖性;胃复方含药血浆可能通过上调PPARγ蛋白表达及下调PTTG、c-Myc、hTERT蛋白表达来抑制人胃癌BGC-823细胞增殖,从而诱导细胞凋亡。
Objective To explore the effect of weifufang medicated-plasma on BGC-823 proliferation in human gastric cancer on the base of PPARγ | PTTG pathway. Methods Weifufang medicated-plasma was prepared and exerted to BGC-823 proliferation in human gastric cancer with different concentrations. Separately,in 24 h,48 h and 72 h,MTS method was adopted to determine the inhibition rate of cell proliferation. The half-number inhibition rate of IC_(50) was calculated. Using weifufang medicated-plasma on BGC-823,with 1/4 IC_(50) concentration,72 h later,the flow cytometry was applied to determine the changes in cell apoptosis. Western blot method was used to determine the protein expressions of PPARγ,PTTG,c-Myc and h TERT in BGC-823 in human gastric cancer. Results The weifufang medicated-plasma displayed the obvious inhibition on BGC-823 proliferation in human gastric cancer and the dose-time dependence was presented. In comparison among the interventions with 5%,10% and 20% medication,the differences were significant(P< 0. 05). In the medication groups with the concentration over 10%,the cell growth mostly stopped. The differences were not significant among the groups in 24 h and 48 h(P > 0. 05),and the differences were significant among groups in 72 h(P< 0. 05). In the low-dose,middle-dose and high-dose groups of weifufang medicated-plasma,IC_(50) was 30. 32%,19. 37% and 14. 69% successively. Compared with the blank group and PBS negative control group,the apoptosis rate in the weifufang group was increased(P < 0. 01). In the middle-dose group,after intervention with IC_(50) drug-contained plasma for 72 h,compared with the control group,weifufang medicated-plasma increased the protein expression of PPARγ(P < 0. 05,P < 0. 01) and down-regulated the protein expressions of PTTG,c-Myc and h TERT(P < 0. 01). Conclusion The weifufang medicated-plasma presents the inhibition on BGC-823 proliferation in human gastric cancer,indicating the time-concentration dependence. Weifufang medicated-plasma suppresses BGC-823 proliferation through the up-regulation of PPARγ protein expression and the down-regulation of the protein expressions of PTTG,c-Myc and h TERT so as to induce cell apoptosis.
Objective To explore the effect of weifufang medicated-plasma on BGC-823 proliferation in human gastric cancer on the base of PPARγ | PTTG pathway. Methods Weifufang medicated-plasma was prepared and exerted to BGC-823 proliferation in human gastric cancer with different concentrations. Separately,in 24 h,48 h and 72 h,MTS method was adopted to determine the inhibition rate of cell proliferation. The half-number inhibition rate of IC_(50) was calculated. Using weifufang medicated-plasma on BGC-823,with 1/4 IC_(50) concentration,72 h later,the flow cytometry was applied to determine the changes in cell apoptosis. Western blot method was used to determine the protein expressions of PPARγ,PTTG,c-Myc and h TERT in BGC-823 in human gastric cancer. Results The weifufang medicated-plasma displayed the obvious inhibition on BGC-823 proliferation in human gastric cancer and the dose-time dependence was presented. In comparison among the interventions with 5%,10% and 20% medication,the differences were significant(P< 0. 05). In the medication groups with the concentration over 10%,the cell growth mostly stopped. The differences were not significant among the groups in 24 h and 48 h(P > 0. 05),and the differences were significant among groups in 72 h(P< 0. 05). In the low-dose,middle-dose and high-dose groups of weifufang medicated-plasma,IC_(50) was 30. 32%,19. 37% and 14. 69% successively. Compared with the blank group and PBS negative control group,the apoptosis rate in the weifufang group was increased(P < 0. 01). In the middle-dose group,after intervention with IC_(50) drug-contained plasma for 72 h,compared with the control group,weifufang medicated-plasma increased the protein expression of PPARγ(P < 0. 05,P < 0. 01) and down-regulated the protein expressions of PTTG,c-Myc and h TERT(P < 0. 01). Conclusion The weifufang medicated-plasma presents the inhibition on BGC-823 proliferation in human gastric cancer,indicating the time-concentration dependence. Weifufang medicated-plasma suppresses BGC-823 proliferation through the up-regulation of PPARγ protein expression and the down-regulation of the protein expressions of PTTG,c-Myc and h TERT so as to induce cell apoptosis.
引文
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[17]沈云龙,安泰学,罗巧灵,等.慢病毒介导shRNA抑制垂体腺瘤PTTG基因表达及对细胞增殖和侵袭能力的影响[J].中山大学学报:医学科学版,2016,37(4):522-529.
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[19]Kim T,Cui R,Jeon Y J,et al.MYC-repressed long noncoding RNAs antagonize MYC-induced cell proliferation and cell cycle progression[J].Oncotarget,2015,6(22):18780-18789.
[20]潘华峰,赵自明,蔡甜甜,等.胃痞消阻断Notch/DLLs通路抑制胃癌前病变大鼠Lgr5胃癌干细胞增殖与侵袭迁移的分子机制研究[J].中华中医药杂志,2017,32(2):523-528.
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[2]Guo F,Ren X,Dong Y,et al.Constitutive expression of PPARγinhibts proliferation and migeation of gastric cancer cells and downregulates Wnt-Catenin signaling pathway downstream target genes TERT and ENAH[J].Gene,2016,584(1):31-37.
[3]Cao LQ,Shao ZL,Liang HH,et al.Activation of peroxisome proliferator-activated receptor-γ(PPARγ)inhibits hepatoma cell growth via downregulation of SEPT2 expression[J].Cancer Lett,2015,359(1):127-135.
[4]魏刚.标准汤剂论[J].中国中医药信息杂志,1998,5(4):6-8.
[5]Siegel R,Ma J,Zou Z,et al.Cancer statistics[J].CA Cancer J Clin,2014,64(1):9-29.
[6]Oba K,Paoletti X,Bang YJ,et al.Role of chemotherapy for advancerd/recurrent gastric cancer:an individual-patient-data meta analysis[J].Eur J Cancer,2013,49(7):1565-1577.
[7]吴军,张俊玲,王婧,等.黄芪多糖通过抑制磷酸化蛋白激酶B表达增强阿帕替尼抗胃癌细胞作用的研究[J].中国医院用药评价与分析,2017,17(6):731-734.
[8]陈雪梅,杨焕芝,施敏,等.长柱重楼总皂苷的抗肿瘤活性及急性毒性作用研究[J].药物评价研究,2017,40(7):904-910.
[9]徐卫东,芮梦珏,欧阳臻,等.术属植物水提物对胃癌SGC-7901细胞的增殖抑制作用研究[J].时珍国医国药,2015,26(12):2861-2864.
[10]罗金强,刘宏斌.半枝莲、白花蛇舌草抗肿瘤的研究进展[J].现代肿瘤医学,2014,22(2):481-484.
[11]王晓菲,刘春琰,窦德强.中药茯苓抗肿瘤有效组分研究[J].辽宁中医杂志,2014,41(6):1240-1243.
[12]吉爱军,陈建伟,刘沈林,等.三棱散对人胃癌SGC-7901细胞增殖作用的影响[J].辽宁中医药杂志,2016,43(1):114-117.
[13]唐德才,臧文华,冯海红.莪术不同品种含药血清对人胃癌细胞BGC823增殖、凋亡及核质比的影响[J].北京中医药大学学报,2013,36(4):254-257.
[14]Bojkova B,Kajo K,Garajova M,et al.Rosiglitazone shows Partial oncostatic etffect in rat mammary carcinogen esis[J].Neoplasm,2013,60(1):46-55.
[15]胡亚,李东芳.胃复方含药血浆对胃癌SGC-7901细胞株增殖及PPARγ基因活性的影响[J].中国中西医结合消化杂志,2016,24(3):178-181.
[16]梁华晟,秦映芬,罗佐杰.PPARγ与内分泌肿瘤关系的研究进展[J].陕西医学杂志,2006,35(7):865-866,880.
[17]沈云龙,安泰学,罗巧灵,等.慢病毒介导shRNA抑制垂体腺瘤PTTG基因表达及对细胞增殖和侵袭能力的影响[J].中山大学学报:医学科学版,2016,37(4):522-529.
[18]Suojun Z,Feng W,Dongsheng G,et al.Targeting Raf/MEK/ERKpathway in pituitary adenomas[J].Eur J Cancer,2012,48(3):389-395.
[19]Kim T,Cui R,Jeon Y J,et al.MYC-repressed long noncoding RNAs antagonize MYC-induced cell proliferation and cell cycle progression[J].Oncotarget,2015,6(22):18780-18789.
[20]潘华峰,赵自明,蔡甜甜,等.胃痞消阻断Notch/DLLs通路抑制胃癌前病变大鼠Lgr5胃癌干细胞增殖与侵袭迁移的分子机制研究[J].中华中医药杂志,2017,32(2):523-528.
[21]Albihn A,Johnsen J I,Arsenian Henriksson M.MYC in oncogenesis and as a target for cancer therapies[J].Advances in cancer research,2010(107):163-224.