论文部分内容阅读
目的:研究RNA干扰人抗原R(human antigen R,HuR)基因的表达对人乳腺癌耐药细胞株MCF-7/Adr对多柔比星(doxorubicin)敏感性的影响。方法:构建靶向HuR基因的shRNA表达质粒(pGenesil-siHuR),稳定转染至MCF-7/Adr细胞,real-time PCR检测细胞中MDR1 mRNA的表达,Western blotting检测MCF-7/Adr细胞中由MDR1基因编码的P糖蛋白(P-glycoprotein,P-gp)的表达,MTT法检测pGenesil-siHuR转染后MCF-7/Adr细胞在多柔比星作用后的存活率和IC50,流式细胞术检测MCF-7/Adr细胞的凋亡率。结果:与未转染的MCF-7/Adr细胞比较,pGenesil-siHuR质粒转染MCF-7/Adr细胞中MDR1 mRNA的表达水平明显减低[(0.184±0.029)vs(1.203±0.026),P<0.01],P-gp表达水平明显降低。pGenesil-siHuR质粒转染MCF-7/Adr细胞后,MCF-7/Adr细胞对多柔比星的IC50从未转染的(148.2±2.3)nmol/L降至(42.9±0.4)nmol/L;经多柔比星处理后,pGenesil-siHuR质粒转染组MCF-7/Adr细胞的凋亡率明显上升[(34.6±1.1)%vs(1.1±0.2)%,P<0.01]。结论:RNA干扰HuR的表达能抑制MDR1基因的表达,增加耐药乳腺癌MCF-7/Adr细胞对多柔比星的敏感性。
AIM: To investigate the effect of human interferor RNA (HuR) gene expression on the sensitivity of human breast cancer cell line MCF-7 / Adr to doxorubicin. Methods: shRNA expression plasmid targeting HuR gene (pGenesil-siHuR) was constructed and stably transfected into MCF-7 / Adr cells. Real-time PCR was used to detect the expression of MDR1 mRNA in MCF-7 / Adr cells The expression of P-glycoprotein (P-glycoprotein) was detected by MDR1 gene. The survival rate and IC50 of MCF-7 / Adr cells after transfection with pGenesil-siHuR were determined by MTT assay. Cytometry was used to detect the apoptosis rate of MCF-7 / Adr cells. Results: The expression of MDR1 mRNA in MCF-7 / Adr cells transfected with pGenesil-siHuR plasmid was significantly lower than that in untransfected MCF-7 / Adr cells [(0.184 ± 0.029) vs (1.203 ± 0.026, P < 0.01], P-gp expression was significantly lower. The IC50 of doxorubicin in MCF-7 / Adr cells transfected with pGenesil-siHuR plasmid was decreased from (148.2 ± 2.3) nmol / L to (42.9 ± 0.4) nmol / L ; After treated with doxorubicin, the apoptosis rate of MCF-7 / Adr cells transfected with pGenesil-siHuR plasmid was significantly increased [(34.6 ± 1.1)% vs (1.1 ± 0.2)%, P <0.01]. Conclusion: The expression of HuR induced by RNAi can inhibit the expression of MDR1 gene and increase the susceptibility of multidrug-resistant breast cancer MCF-7 / Adr cells to doxorubicin.