miRNA21对人肺癌A549细胞增殖及迁移的影响

The effects of miRNA21 on the proliferation and migration of human lung cancer cell line A549 cells

目的:探讨抑制miRNA21对人肺癌A549细胞体外增殖、迁移及凋亡的影响。方法:采用体外转染法将miRNA21-5p瞬时转染A549细胞后,应用Real-time PCR特异探针法检测人肺癌细胞系中miRNA21的表达情况;应用MTT法检测人肺癌A549细胞的增殖情况;应用Transwell小室检测人肺癌A549细胞的迁移情况;流式细胞术(flow cytometry,FCM)检测细胞的凋亡情况;Western blot检测人肺癌A549细胞中表皮因子生长受体(epidermal growth factor receptor,EGFR)的表达。结果:与正常对照组和miRNA21-NC阴性对照组相比,miRNA21-5p转染组人肺癌A549细胞中miRNA21表达水平显著降低,EGFR表达显著下调,细胞生长显著受抑制,且促进人肺癌A549细胞的凋亡(P<0.05)。结论:抑制miRNA21在人肺癌A549细胞中的表达,能够抑制A549细胞的生长,促进其凋亡,miRNA21有可能作为治疗肺癌的新靶标之一。

Objective： To explore the effects of miRNA21 on the proliferation and migration of human lung cancer cell line A549 in vitro. Methods： miRNA21- 5p was transiently transferred into A549 cells using FUGENE- HD reagent in vitro. The expression level of miRNA21 in A549 cells was determined using specific probe of Real- time PCR before and after transfection. The proliferation of A549 cells was accessed using MTT assay. The migration of A549 cells was accessed using Transwell. The cell cycle was analyzed by FCM. The expression of epidermal growth factor receptor was determined by Western blot. Results： The expression level of miRNA21 significantly decreased in miRNA21- 5p transfected A549 cells compared with that in normal control group and miRNA21 negative control group（ P〈0. 05）. Similarly,the expression of EGFR was markedly down- regulated,and the proliferation was significantly inhibited（ P〈0. 05）. Conclusion： Inhibiting miRNA21 may decrease the proliferation and migration of A549 cells,suggesting that miRNA21 might be one of novel targets for the biological therapy of human lung cancer.