| RNA干扰GPC3基因表达影响肝癌细胞Huh-7凋亡的机制研究 |
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| 引用本文： | 郑大勇,阮健,刘斐烨,何本夫,黄娜,罗西,秦如斋,罗荣城.RNA干扰GPC3基因表达影响肝癌细胞Huh-7凋亡的机制研究[J].热带医学杂志,2011,11(5):516-519. |
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| 作者姓名： | 郑大勇 阮健 刘斐烨 何本夫 黄娜 罗西 秦如斋 罗荣城 |
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| 作者单位： | 南方医科大学南方医院肿瘤中心,广东,广州,510515 |
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| 摘 要： | 目的观察RNA干扰GPC3基因表达对人肝癌细胞Huh-7凋亡的影响,并探讨其初步机制。方法设计并合成靶向GPC3的特异性siRNA片段,通过脂质体转染法瞬时转染肝癌细胞Huh-7,48h后验证siRNA的干扰效率;Annexin V/PI染色法观察GPC3基因沉默对细胞凋亡的影响;通过Western blot和定量PCR来检测caspase3的蛋白水平和核酸水平表达。结果设计合成的GPC3siRNA转染后,能够有效抑制Huh-7细胞中GPC3的表达;流式细胞仪检测结果显示,GPC3干扰组细胞在转染后48、72h的凋亡率显著高于空白对照组(P<0.01),96h后两组凋亡率差异无统计学意义(P>0.05)。GPC3干扰组的caspase3在转染后48、72h的表达高于未转染组,96h后两组间caspase3水平差异无统计学意义(P>0.05)。结论 siRNA静默GPC3基因能促进肝癌细胞凋亡,其机理可能与上调caspase3有关。进而推测,GPC3基因可能成为肝癌靶向治疗的一个新靶点。
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| 关 键 词： | 肝癌 RNA干扰 细胞凋亡 GPC3 caspase-3 |
Inhibition of GPC3 expression via RNA interference influences the apoptosis of a human hepatocellular carcinoma cell line Huh-7 |
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| ZHENG Da-yong,RUAN Jian,LIU Fei-ye,HE Ben-fu,HUANG Na,LUO Xi,QIN Ru-zhai,LUO Rong-cheng.Inhibition of GPC3 expression via RNA interference influences the apoptosis of a human hepatocellular carcinoma cell line Huh-7[J].Journal Of Tropical Medicine,2011,11(5):516-519. |
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| Authors: | ZHENG Da-yong RUAN Jian LIU Fei-ye HE Ben-fu HUANG Na LUO Xi QIN Ru-zhai LUO Rong-cheng |
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| Institution: | ZHENG Da-yong,RUAN Jian,LIU Fei-ye,HE Ben-fu,HUANG Na,LUO Xi,QIN Ru-zhai,LUO Rong-cheng(Oncology Center,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China) |
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| Abstract: | Objective To investigate the effect of RNA knockdown of the GPC3 gene on the apoptosis of hepatocellular carcinoma Huh-7 cells.Methods siRNA targeting GPC3 was designed and synthesized,and then transfected into the Huh-7 cells via Lipofectamine 2000 mediation.The cell apoptotic rate was analyzed by using flow cytometry.Expression of capase3 was detected by RT-PCR and western blot.Results GPC3-targeted siRNA could down-regulate the GPC3 expression of Huh-7.Compared with control group,cell apoptosis rates wer... |
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| Keywords: | GPC3 caspase-3 |
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