论文部分内容阅读
目的探讨腺病毒介导 p53基因(Ad-p53)对人乳腺癌阿霉素(ADM)耐药细胞株MCF-7/ADM 的耐药性及多药耐药基因1(MDR1)的影响。方法以 Ad-p53转染 MCF-7/ADM 细胞株,流式细胞术检测 p53蛋白变化;锥虫蓝活细胞拒染法观察细胞生长情况;MTT 法观察 MCF-7/ADM 细胞对阿霉素耐药性的变化,实时荧光 RT-PCR 法检测 MDR1 mRNA 变化。结果流式细胞术观察到 MCF-7/ADM 细胞经 MOI_(50)的 Ad-p53作用48 h 后,p53蛋白表达率由10.24%升高到36.20%;细胞抑制率为7.4%(P=0.003);逆转 MCF-7/ADM 对 ADM 的耐药倍数11.6倍(P=0.001);实时荧光 RT-PCR 检测结果显示 MDR1 mRNA 相对表达量由1.25±0.01下降至0.91±0.01(P=0.011)。结论腺病毒介导 p53基因能抑制 MCF-7/ADM 细胞 MDR1基因的表达,并能部分逆转MCF-7/ADM 的耐药性,增加阿霉素化疗敏感性。
Objective To investigate the effect of adenovirus-mediated p53 gene (Ad-p53) on the drug resistance of human breast cancer ADM cell line MCF-7 / ADM and multidrug resistance gene 1 (MDR1). Methods MCF-7 / ADM cells were transfected with Ad-p53 and the expression of p53 was detected by flow cytometry. The growth of cells was detected by trypan blue exclusion assay. The proliferation of MCF-7 / ADM cells was evaluated by MTT assay. Drug resistance changes, real-time fluorescence RT-PCR assay MDR1 mRNA changes. Results The expression of p53 protein was up-regulated from 10.24% to 36.20% in 48 h after MOI_ (50) Ad-p53 treatment by flow cytometry. The cell inhibition rate was 7.4% (P = 0.003 (P = 0.001). The real-time RT-PCR results showed that the relative expression of MDR1 mRNA decreased from 1.25 ± 0.01 to 0.91 ± 0.01 (P = 0.011). Conclusion Adenovirus-mediated p53 gene can inhibit the expression of MDR1 gene in MCF-7 / ADM cells and partially reverse the drug resistance of MCF-7 / ADM cells and increase the chemosensitivity of doxorubicin.