RNA干扰HO-1表达对肺腺癌A549细胞生物学行为的影响研究

Effect of silencing HO-1 gene expression on biological behavior of lung adenocarcinoma A549 cells

目的应用RNA干扰技术特异性抑制血红蛋白加氧酶-1(HO-1)的表达,观察HO-1对肺腺癌A549细胞增殖、凋亡及侵袭能力的影响。方法将体外构建的HO-1小分子RNA(siRNA)转染入肺腺癌A549细胞,分为空白对照组(blank组)、脂质体组(mock组)、阴性对照组(NC组)、HO-1 siRNA组;应用实时荧光定量PCR、蛋白质印迹法检测HO-1 mRNA和蛋白表达水平;分别以CCK-8法、流式细胞术检测细胞增殖能力和凋亡率,用Transwell试验检测细胞迁移能力。结果细胞培养48、72h后,与blank组、mock组、NC组比较,HO-1 si RNA组细胞存活率均降低(均P<0.05),细胞凋亡率均增高(均P<0.05),G_0期/G_1期细胞比例均增高(均P<0.05),Transwell试验中穿膜细胞数均减少(均P<0.05)。结论 RNA干扰HO-1基因表达后能有效调控肺腺癌A549细胞的恶性生物学行为,抑制肺腺癌A549细胞的增殖,促进凋亡,降低细胞的侵袭能力。

Objective To investigate the effects of silencing heme oxygenase- 1（HO- 1） gene expression by RNA interference（RNAi） on proliferation and invasiveness of human lung adenocarcinoma A549 cells in vitro. Methods The small interfering RNA（ si RNA） targeting HO- 1 gene was constructed and transfected into A549 cells. Reverse transcriptionpolymerase chain reaction（RT- PCR） and Western blot was used to detect the silencing effect of HO- 1 expression. The assays of cell counting kit- 8（CCK- 8）, flow cytometry and Transwell were performed to assess the malignant phenotypes of transfected A549 cells. Results The proliferation of interference group was markedly decreased（P〈0.05）. The apoptosis rate was significant higher in the interference group（P0.05）, and the number of cells in G_o/G_1 phase was markedly increased（P0.05）.The number of migrated cells in interference group was significant higher than those in non- transfected control groups（P0.05）. Conclusion Silencing HO- 1 gene expression by RNA interference can significantly induce cell apoptosis, and inhibit the proliferation and migratory capacity of lung adenocarcinoma A549 cells in vitro.