摘要
目的探讨非整倍体结肠癌对顺铂的耐药性。方法结肠癌细胞HCT116经多西环素(Dox)诱导为敲低MAD2表达的为Dox(+)组(非整倍体),未经Dox处理的为Dox(-)组(整倍体)。顺铂处理2组细胞48 h,以CCK-8法和细胞计数法分析顺铂对细胞生长的影响。Western blot法检测顺铂对细胞造成的凋亡影响。实时荧光定量即时聚合酶链锁反应(qRT-PCR)实验检测细胞中BAX、PUMA和NOXA凋亡基因表达情况。结果以不同浓度顺铂处理细胞,通过CCK-8实验和细胞计数实验检测发现,Dox(+)组细胞较Dox(-)组细胞对顺铂具有耐药性。Western Blot实验发现,Dox(+)组细胞的耐药性与其经顺铂处理后凋亡蛋白表达较低有关。qRT-PCR实验验证顺铂处理后Dox(+)组细胞较Dox(-)组细胞凋亡基因PUMA、NOXA、BAX表达显著减少(P<0.05或P<0.01)。结论非整倍体结肠癌细胞对顺铂具有耐药性。
Objective To explore drug resistance of aneuploid colon cancer cells to cisplatin.Methods The HCT116 cells with expression of knockdown MAD2 induced by doxycycline(DOX)were designed as DOX(+)group(aneuploid),and HCT116 cells without process of DOX were designed as DOX(-)group(euploid).CCK-8 method was used to process cells in both groups,and CCK-8 test and cytometry were used to analyze the effect of cisplatin on cell growth.Quantitative real time polymerase chain reaction(qRT-PCR)was used to detect the expression of pro-apoptotic genes BAX,PUMA and NOXA in the cells.Results Cells were treated with cisplatin at different concentrations.CCK-8 test and cytometry showed that cells in DOX(+)group were more resistant to cisplatin than those in DOX(-)group.Western blot showed that the resistance of cells in DOX(+)group was related to the lower expression of apoptotic protein after cisplatin treatment.The qRT-PCR showed that the expressions of apoptosis gene PUMA,NOXA and BAX in DOX(+)group were significantly lower than those in DOX(-)group(P<0.05 or P<0.01)after process of cisplatin.Conclusion The aneuploid colon cancer cells are resistant to cisplatin treatment.
作者
张汉卿
刘颖
房晓
ZHANG Hanqing;LIU Ying;FANG Xiao(Medical College of Yangzhou University,Yangzhou,Jiangsu,225001;Clinical Medical College of Yangzhou University,Yangzhou,Jiangsu,225001)
出处
《实用临床医药杂志》
CAS
2020年第1期42-45,共4页
Journal of Clinical Medicine in Practice
基金
国家自然科学基金资助项目(81402482)
江苏省青年医学重点人才培养项目(QNRC2016322)
关键词
顺铂
非整倍体
结肠癌细胞
耐药性
cisplatin
aneuploidy
colon cancer cells
drug resistance
