【摘要】 目的探讨慢病毒短发夹RNA(shRNA)介导的多形性腺瘤基因样蛋白2(PLAGL2)沉默对肝癌细胞恶性行为的影响及其机制。方法 Real-time PCR与Western blotting分别检测肝癌组织及癌旁组织中PLAGL2的表达水平;体外培养肝癌细胞MHCC97-L,构建慢病毒载体质粒PLAGL2-shRNA与对照NC-shRNA,转染MHCC97-L细胞,采用嘌呤霉素筛选稳转株;CCK-8、Transwell小室实验检测沉默PLAGL2后MHCC97-L细胞的增殖活力以及迁移和侵袭数量;Western blotting检测p-PI3K和p-Akt蛋白表达。用PI3K/Akt信号通路激活剂处理MHCC97-L细胞,检测细胞增殖、迁移及侵袭能力。结果肝癌组织中PLAGL2表达水平显著升高(P<0.05);9株MHCC97-L细胞转染PLAGL2-shRNA可明显降低PLAGL2表达水平,同时,细胞增殖、迁移、侵袭能力减弱(P<0.05),p-PI3K、 p-Akt表达受抑制(P<0.05);PI3K/Akt激活剂可明显逆转上述现象。结论慢病毒shRNA载体介导的PLAGL2沉默,可显著抑制肝癌细胞恶性行为,其作用机制可能与抑制PI3K/Akt信号通路的激活有关。更多还原

【Abstract】 Objective To investigate the effect of short hairpin RNA(shRNA)-mediated pleomorphic adenoma gene like-2(PALAG2) silencing on the malignant behavior of hepatocellular carcinoma cells and its mechanism. Methods Real-time PCR and Western blotting were used to detect the expression level of PLAGL2 in liver cancer tissues and adjacent tissues. Hepatoma cells MHCC97-L were cultured in vitro, the lentiviral vector plasmid PLAGL2-shRNA and control NC-shRNA were constructed, transfected into MHCC97-L cells, and stable transfected strains were selected with puromycin. CCK-8 and Transwell chamber assay detected the proliferation activity and the number of migration and invasion of MHCC97-L cells after silencing PLAGL2. Western blotting was used to detect the expression of p-PI3 K and p-Akt proteins. The PI3 K/Akt signaling pathway activator was used to treat MHCC97-L cells to detect cell proliferation, migration and invasion. Results The expression of PLAGL2 was significantly increased in liver cancer tissue(P<0.05). Transfection of 9 strains of MHCC97-L cells with PLAGL2-shRNA could significantly reduce the expression level of PLAGL2, and the ability of proliferation, migration, and invasion of MHCC97-L cells was also weakened(P<0.05), and the expression levels of p-PI3 K, and p-Akt were inhibited(P<0.05), PI3 K/Akt activator could obviously reverse the above phenomenon. Conclusion shRNA lentiviral vector pathway can effectively silence the expression of PLAGL2 gene in hepatocarcinoma cells. Silencing of PLAGL2 can significantly inhibit the malignant behavior of proliferation, migration and invasion of hepatocarcinoma cells, and its mechanism may be related to the inhibition of PI3 K/Akt signaling pathway activation.更多还原