过表达hsa-miR-202对非小细胞肺癌顺铂耐药细胞生物学行为的影响

摘要

目的 探讨过表达hsa-miR-202对非小细胞肺癌(NSCLC)顺铂耐药A549/DDP细胞(以下称A549/DDP细胞)生物学行为的影响.方法 体外传代培养A549/DDP细胞,取传3代对数生长期细胞,随机分为空白对照组、阴性对照组、hsa-miR-202组.阴性对照组、hsa-miR-202组分别转染mimic NC、hsa-miR-202 mimic,空白对照组不予转染.采用MTT法检测各组转染24 h再培养24、48、72、96、120 h细胞增殖率,采用平板集落形成实验检测各组转染24 h细胞集落形成个数,采用流式细胞术检测各组转染24 h细胞周期,采用FITC Annexin V/PI双染法检测各组转染24 h细胞凋亡率.结果 三组转染24 h再培养24、48 h细胞增殖率比较P均>0.05;hsa-miR-202组转染24 h再培养72、96、120 h细胞增殖率明显低于空白对照组和阴性对照组(P均<0.05),空白对照组与阴性对照组比较P>0.05.hsa-miR-202组细胞集落形成个数明显少于空白对照组和阴性对照组(P均<0.05),空白对照组与阴性对照组比较P>0.05.hsa-miR-202组G1期所占比例明显高于空白对照组和阴性对照组,S期、G2期所占比例明显低于空白对照组和阴性对照组,空白对照组与阴性对照组各期细胞所占比例比较P均>0.05.hsa-miR-202组细胞凋亡率明显高于空白对照组和阴性对照组(P均<0.05),空白对照组与阴性对照组比较P>0.05.结论 过表达hsa-miR-202能够抑制A549/DDP细胞增殖并促进其凋亡.%Objective To investigate the effect of hsa-miR-202 on the biological behavior of cisplatin-resistant A549/ DDP cells (hereinafter referred to as A549/DDP cells) of non-small-cell lung cancer (NSCLC). Methods A549/DDP cells were subcultured and the cells of passage 3 in the logarithmic growth phase were inoculated on 6-well plate and randomly divided into the blank control group, negative control group, and hsa-miR-202 group. Cells in the negative control group and hsa-miR-202 group were transfected with mimic NC and hsa-miR-202 mimic, respectively, and the blank control group was not transfected. The cells were collected at 24 h after transfection and the proliferation rate of the cells was detected by MTT assay at 24, 48, 72, 96 and 120 h after transfection. The number of colony formation was measured by plate colony formation assay. The cell cycle was detected by flow cytometry, and the apoptosis rate was detected by FITC Annexin V/PI double staining. Results The cell proliferation rate in the hsa-miR-202 group was significantly lower than that in the blank control group and negative control group (P＜0. 05); the number of colony forming in the hsa-miR-202 group was significantly lower than that in the blank control group and negative control group (P＜0. 05); the proportion of cells in the G1 phase of the hsa-miR-202 group was significantly higher than that in blank control group and negative control group, and the proportion of cells in S phase and G2 phase was significantly lower than that in the blank control group and negative control group; the apoptosis rate of the hsa-miR-202 group was significantly higher than that of the blank control group and the negative control group (P < 0. 05); no significant differences were found in the above parameters between the blank control group and negative control group (all P > 0. 05). Conclusion Overexpression of hsa-miR-202 can inhibit the pro-liferation of A 549/DDP cells and promote their apoptosis.