RNA干扰沉默mTOR基因对肝癌HepG2细胞侵袭和转移的影响(1)
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[摘要] 目的：探讨RNA干扰技术对人肝癌HepG2细胞mTOR的表达及细胞侵袭与转移的影响。方法：体外合成mTOR siRNA，并转染HepG2细胞24 h，同时设无义对照组(转染无义对照siRNA)、空白对照组(转染空脂质体)及正常对照组(不转染)。采用western blot法检测各组HepG2细胞的mTOR蛋白表达；应用Transwell小室细胞体外实验检测转染后肝癌HepG2细胞的穿膜细胞数，评价肝癌细胞体外侵袭和转移能力的变化。结果：mTOR siRNA转染组HepG2细胞mTOR蛋白的表达低于各对照组(P<0.05)；HepG2细胞侵袭和转移力均显著低于各对照组(P<0.05)。结论：mTOR siRNA可有效抑制HepG2细胞mTOR蛋白的表达，且能抑制HepG2细胞的侵袭和迁移。
[关键词] 原发性肝癌 HepG2细胞 mTOR RNA干扰 HepG2细胞 侵袭 转移
[中图分类号] R735.7 [文献标识码]A [文章编号]1673-7210(2011)02(c)-014-03
Effect of mTOR gene inhibition by siRNA on invasion and metastasis of human Hepatoma HepG2 cells
GU Liang, ZHANG Yangde*
(Ministry of Health, Central South University, Hepatobiliary & Enteric Surgery Research Center, Changsha City, Hu′nan Province, Changsha410008, China)
[Abstract] Objective: To study the effect of mTOR siRNA on the expression of mTOR gene, cell invasion and metastasis of human Hepatoma HepG2 cell. Methods: The oligonucleotide templates of mTOR siRNA were designed primarily, then were used to transfect HepG2 with 24 h, and the control groups were established accordingly by using siRNA with insignificant order, liposome only and no transfection. The level of protein of mTOR was measured by western blot. Transwell chamber in vitro was to detect the cell count which were able to invade matrigel membrane to assess the ability of cell invasion and metastasis. Results: The expression of mTOR protein in the groups transfected with mTOR siRNA was lower than that of the control groups (P<0.05). The invasion and metastasis of HepG2 cells transfected with mTOR siRNA were decreased than those of the control groups (P<0.05). Conclusion: RNA interference of mTOR in HepG2 cell line could effetively restrain the expression of mTOR, which could significantly inhibit the invasion and metastasis of HepG2 cell.
[Key words] Hepatocellular carcinoma; HepG2 cells; mTOR; RNA interference; Invasion; Metastasis
原发性肝癌是原发于肝细胞或肝内胆管上皮细胞的恶性肿瘤之一。肝癌起病隐匿，发展迅速，恶性程度高，一旦发现多为中晚期，不论是手术还是姑息治疗，效果均不理想。由于肝癌是一种高侵袭、高转移性癌症，临床治疗又易复发转移，使疗效难以提高。据统计 ......
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