摘要
目的:构建奥沙利铂耐药结肠癌细胞株(SW480/ADR),观察miRNA-34a对SW480/ADR细胞奥沙利铂敏感性的影响。方法:使用浓度梯度法构建奥沙利铂耐药细胞株(SW480/ADR)。miR-34a慢病毒转染SW480/ADR细胞,MTT法检测细胞增殖,Real-time PCR定量检测miR-34a表达,Western blot检测蛋白表达。结果:SW480、SW480/ADR和过表达miR-34a-SW480/ADR细胞miR-34a的2-△△Ct值为0.15±0.03、0.12±0.02和0.74±0.13,各组间差异有统计学意义(P<0.01)。奥沙利铂对SW480、SW480/ADR和过表达miR-34a-SW480/ADR细胞的IC50值分别为8.64±1.54、24.16±3.72和11.52±1.22μmol/L,各组间差异有统计学意义(P<0.01)。SW480、SW480/ADR和过表达miR-34a-SW480/ADR细胞OCT-4蛋白的相对灰度值为0.096±0.014、0.352±0.053和0.136±0.017,各组间差异有统计学意义(P<0.01)。结论:miR-34a可以显著降低结肠癌奥沙利铂耐药细胞OCT-4蛋白表达,逆转结肠癌细胞对奥沙利铂的耐药。
Objective:To establish oxaliplatin-resistant colon cancer cell line(SW480/ADR)and observe the change of sensitivity of oxaliplatin in microRNA-34a-overexpression SW480/ADR cells.Methods:The SW480/ADR cell line was established by concentration gradient method.SW480/ADR cells were transfected by miRNA-34a lentivirus vector.mi R-34a was detected by Real-time PCR.The cell proliferation was completed by MTT analysis.The protein expression was determined by western blot. Results: 2-△△Ct values of miR-34a in SW480, SW480/ADR, and miR-34a overexpression SW480/ADR cells was 0.15±0.03, 0.12±0.02, and 0.74±0.13. There were significant differences among the three groups(P<0.01). The IC50 values of oxaliplatin to SW480, SW480/ADR, and miR-34a overexpression SW480/ADR cells was 8.64±1.54, 24.16±3.72, and 11.52±1.22 μmol/L. There were significant differences among the three groups(P<0.01). The protein expression of OCT-4 in SW480, SW480/ADR, and miR-34a overexpression SW480/ADR cells was 0.096±0.014, 0.352±0.053, and 0.136±0.017. There were significant differences among the three groups(P<0.01). Conclusion: miR-34a can significantly reduce OCT-4 expression in oxaliplatin-resistant colon cancer cells and reverse the resistance of colon cancer cells to oxaliplatin.
作者
田由京
陈兴超
张浩
李军
李合
TIAN You-jing;CHEN Xing-chao;ZHANG Hao;LI Jun;LI He(Department of General Surgery,First Affiliated Hospital of Hainan Medical College,Haikou 570000,China)
出处
《中国现代普通外科进展》
CAS
2020年第2期89-92,共4页
Chinese Journal of Current Advances in General Surgery
基金
2015年海南自然科学基金项目(20158351)。
