| 摘要: |
| 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-1siRNA组；应用实时荧光定量PCR、蛋白质印迹法检测HO-1mRNA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-1siRNA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），G0期/G1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。 |
| 关键词: 肺肿瘤 HO-1 RNA 干扰 |
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| Effect of silencing HO-1 gene expression on biological behavior of lung adenocarcinoma A549 cells |
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XIA Lilong, JU Sheng, YUAN Feng, ZHU Xinhai, SHU Yue, CHEN Guoping
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Zhejiang Hospital
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| Abstract: |
| 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 ( siRNA ) targeting HO-1 gene was constructed and transfected into A549 cells . Reverse transcription- polymerase 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(P＜0.05), and the number of cells in Go/G1 phase was markedly increased(P＜0.05). The number of migrated cells in interference group was significant higher than those in non-transfected control groups (P＜ 0.05). 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． |
| Key words: Lung neoplasms HO-1 RNA interference |
