文章摘要
蒯兴旺,黄骁辰,唐 奇,赵 薇,章明炯,陈 渊,贾立周,杨婷婷,仇镇宁,朱 进,冯振卿.Trop2对胃癌细胞耐药性的影响及机制研究[J].南京医科大学学报,2018,(7):885~891
Trop2对胃癌细胞耐药性的影响及机制研究
Effects and mechanism of Trop2 on drug resistance of gastric cancer cells
投稿时间:2018-03-30  
DOI:10.7655/NYDXBNS20180704
中文关键词: Trop2  胃癌  耐药  柔红霉素  TopoⅡα
英文关键词: Trop2  gastric cancer  drug resistance  daunorubicin  TopoⅡα
基金项目:国家自然科学基金(81773100)
作者单位
蒯兴旺 南京医科大学病理学系国家卫计委抗体技术重点实验室江苏 南京 211166 
黄骁辰 南京医科大学病理学系国家卫计委抗体技术重点实验室江苏 南京 211166 
唐 奇 国家卫计委抗体技术重点实验室江苏 南京 211166 
赵 薇 南京医科大学附属南京医院病理科江苏 南京 210006 
章明炯 南京医科大学第二附属医院耳鼻喉科,江苏 南京 210011 
陈 渊 南京医科大学第二附属医院消化科江苏 南京 210012 
贾立周 南京医科大学病理学系国家卫计委抗体技术重点实验室江苏 南京 211166 
杨婷婷 南京医科大学病理学系国家卫计委抗体技术重点实验室江苏 南京 211166 
仇镇宁 国家卫计委抗体技术重点实验室江苏 南京 211166 
朱 进 国家卫计委抗体技术重点实验室江苏 南京 211166
华东医学生物技术研究所江苏 南京 210002 
冯振卿 南京医科大学病理学系国家卫计委抗体技术重点实验室江苏 南京 211166 
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中文摘要:
      目的:探讨滋养层细胞表面抗原2(Trop2)对胃癌细胞化疗耐药的影响及机制。方法:应用慢病毒转染技术将Trop2 shRNA转染至BGC823细胞株,通过嘌呤霉素筛选出稳定转染质粒的细胞株;qRT?PCR、Western blot和免疫荧光方法检测质粒干扰效率;CCK?8细胞增殖试验和流式细胞术检测柔红霉素对BGC823细胞株增殖能力和细胞凋亡的影响;qRT?PCR、Western blot检测耐药相关基因TopoⅡα的mRNA和蛋白表达变化。结果:在稳定转染Trop2 shRNA质粒的shTrop2组中,Trop2 mRNA及蛋白表达水平较对照组(未处理)和shNC组(转染空载体质粒)明显下调;在相同药物浓度下,柔红霉素对shTrop2组的增殖抑制率明显高于对照组和shNC组,shTrop2组半数抑制浓度低于对照组和shNC组(P<0.05),shTrop2组中柔红霉素诱导的细胞凋亡率显著高于对照组和shNC组(P<0.05);稳定干扰Trop2表达后耐药相关基因TopoⅡα的表达显著上调。结论:Trop2通过抑制TopoⅡα的表达,可增强胃癌细胞对柔红霉素的耐药性。
英文摘要:
      Objective:To investigate the effects of trophoblast cell?surface antigen 2(Trop2)on drug resistance of gastric cancer and the underlying mechanism. Methods:BGC823 cells were infected with the lentivirus encoding shRNA against Trop2 and puromycin was used to select stable cell lines;The effects of interference were detected by qRT?PCR,Western blot and immunofluorescence;CCK?8 proliferation test and flow cytometric detection were performed to detect the influence of daunorubicin on cell proliferation and apoptosis. The mRNA and protein expressions of TopoⅡα were detected by qRT?PCR and Western blot. Results:The mRNA and protein expressions of Trop2 in the shTrop2 group(transfected with Trop2 shRNA)were significantly lower than those in the control group(untreated)and the shNC group(transfected with normal control shRNA);The shTrop2 group showed higher proliferation inhibition rates after daunorubicin treatment than these in the control group and the shNC group,and the IC50 value of the shTrop2 group was lower than that in the control group and the shNC group(P<0.05);The daunorubicin?induced cell apoptosis rate in the shTrop2 group was higher than that in the control group and the shNC group(P<0.05);The mRNA and protein expressions of drug resistance?related gene TopoⅡα decreased after interference with Trop2 expression stably. Conclusion:Trop2 can intensify daunorubicin resistance of gastric cancer cells by inhibiting the expression of TopoⅡα.
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