miR-200a对肺癌A549细胞增殖、迁移及凋亡的影响

Effect of miR-200a on the proliferation,migration and apoptosis of lung cancer cell line A549

目的探讨miR-200a在肺癌细胞中的表达及其对A549细胞增殖、迁移及凋亡等生物学功能的影响。方法采用RTPCR检测肺癌细胞株A549、H23、H460和永生化人支气管上皮细胞株16HBE的表达水平。采用脂质体转染法将miR-200a mimics和阴性对照序列(miR-negtive control,miR-NC)分别转染A549细胞,运用CCK-8实验和平板克隆形成实验检测细胞增殖能力的改变情况;Transwell实验检测细胞迁移能力;流式细胞术检测细胞凋亡情况。利用生物信息学方法预测miR-200a的靶基因。结果 miR-200a在肺癌细胞株A549、H23和H460相对表达水平均低于16HBE组,差异有统计学意义(P<0.05)。miR-200a mimics和miR-NC组分别转染A549细胞,CCK-8实验显示miR-200a mimics组在第4、5天时的OD值明显低于miRNC组,差异有统计学意义(P<0.05);平板克隆形成实验显示miR-200a mimics组细胞克隆形成率为(33.13±0.74)%,明显少于miR-NC组(45.57±1.27)%,差异有统计学意义(P<0.05)。Transwell实验显示,miR-200a mimics穿膜细胞数为(71.60±17.90)个,少于miR-NC组[(140.20±17.88)个],差异有统计学意义(P<0.05)。流式细胞术检测显示,miR-200a mimics凋亡率为(17.80±1.90)%,明显高于miR-NC组[(11.33±1.96)%],差异有统计学意义(P<0.05)。生物信息学方法预测Tiam1等可能是miR-200a的靶基因。结论 miR-200a能够抑制肺癌细胞A549的增殖、迁移并促进其凋亡,为肺癌的治疗提供潜在靶点。miR-200a可能通过靶基因Tiam1发挥其对肿瘤的调控作用。

Purpose To investigate the expression of miR- 200a in different lung cancer cell lines and its effect on proliferation, migration, and apoptosis in A549 cells. Methods The expressions of miR-200a in different lung cancer cell lines were detected by RT-PCR. miR-200a mimics was transfected into A549 cells by Lipofectamine RNAiMax. The change of proliferation ablility of A549 cells was detected by CCK-8 method and plate clone formation assay. Cell migration was examined by Transwell chamber assay. The flow cytometry was used to examine the changes of apoptosis. The possible target genes of miR-200a were forecasted by bioinformatics tools. Results The results of RT-PCR showed that the expression of miR-200a was significant- ly down-regulated in A549, H23 and H460 cell lines than 16HBE cell line. CCK-8 assay showed that the OD values of the mimics group at 4, and 5 days were significantly lower than those in the negative control （ NC ） group （ P 〈 0. 05 ）. Plate clone formation assay showed rate of colony formation in the mimics group was significantly lower than that in the NC group [（33. 13 ±0.74）% vs （45.57 ±1.27）%, P〈0.051. Transwell migration assay showed that the cell number of mimics group that passed the Transwell membrane was significantly lower than that of the NC group E（71.60 ± 17.90） vs （140.20 ± 17. 88）, P 〈0. 05]. Flow cytometry showed that the apoptosis rate of the mimics group was significantly higher than that of the NC group [（17.80 ± 1.90）% vs （11.33 ± 1.96）%, P 〈 0.05 ]. Tiaml may be one of the target gene of miR-200a. Conclusion miR-200a can inhibit the proliferation and migration, and promote apoptosis of lung cancer A549 cells, suggesting a potential new therapeutic agent for lung cancer cell. MiR-200a may play a function of regulation of tumor development through target gene Tiaml.