摘要
目的:建立耐伊立替康(CPT-11)的人结直肠癌细胞株(HT-29/CPT-11),并研究其生物学特性。方法:采用药物浓度梯度递增间歇诱导法建立人结直肠癌耐药株HT-29/CPT-11;CCK-8法检测CPT-11对亲本细胞HT-29和耐药细胞株HT-29/CPT-11细胞的半数抑制率(IC50);绘制细胞生长曲线,并计算细胞倍增时间;流式细胞术分析细胞周期分布;RT-PCR检测HT-29/CPT-11和HT-29细胞MDR-1 mRNA的表达。结果:(1)成功建立人结直肠癌耐药细胞株HT-29/CPT-11,耐药指数为6.51。(2)HT-29/CPT-11耐药细胞株倍增时间较原代细胞HT-29延长[分别为(41.29±1.69)h和(27.12±2.73)h,P<0.05]。(3)流式细胞术检测细胞周期结果显示,耐药细胞株HT-29/CPT-11 G1期细胞数目增加,S期细胞数目减少(P<0.05)。(4)耐药细胞HT-29/CPT-11的MDR-1 mRNA表达水平明显高于亲本细胞株HT-29,分别为1.086±0.054和0.416±0.02(P<0.05)。结论:成功构建了人结直肠癌耐药细胞株HT-29/CPT-11,为下一步研究耐药机制奠定实验基础。
Objective： To establish a irinotecan（CPT-11） resistance human colorectal cancer cell line and study its biological characteristics,Methods： HT-29/CPT-11 cell line was established by stepwise selection in increasing dose of CPT-11.IC50 of CPT-11 in HT-29 and HT-29/CPT-11 cell lines was detected by CCK-8.Cell growth curve was drawed and the doubling time was accounted.The cell cycle distribution was determined by flow cytometry.The expression levels of MDR-1 mRNA in HT-29 and HT-29/CPT-11 cell lines were examined by RT-PCR.Results：（1） The CPT-11 resistance human colorectal cancer cell line HT-29/CPT-11 was successfully established,the resistance index was 6.51.（2） The doubling time of HT-29/CPT-11 obviously longer than that of HT-29 cell[（41.29±1.69）h vs（27.12±2.73）h,P0.05].（3） Flow cytometry demonstrated that HT-29/CPT-11 cells of phase G1 were increased and S phase were reduced.（4） The expression level of MDR-1 mRNA in HT-29/CPT-11 cell line was higher than that of HT-29（1.086±0.054 vs 0.416±0.02,P0.05）.Conclusion： We have successfully established CPT-11 resistance cell line HT-29/CPT-11,it lays a foundation for the study of resistance mechanism.
出处
《东南大学学报（医学版）》
CAS
2012年第1期31-35,共5页
Journal of Southeast University(Medical Science Edition)
基金
江苏省卫生厅面上科研项目(H200652)
