| GPC3对肝癌细胞株MHCC97-L增殖的影响 |
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| 引用本文： | 秦如斋,刘斐烨,陈斌,谢剑明,杨洋,郑大勇,罗荣城.GPC3对肝癌细胞株MHCC97-L增殖的影响[J].南方医科大学学报,2011,31(3):448-452. |
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| 作者姓名： | 秦如斋 刘斐烨 陈斌 谢剑明 杨洋 郑大勇 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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| 基金项目： | 国家自然科学基金，广东省自然科学基金 |
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| 摘 要： | 目的通过构建磷脂酰肌醇蛋白聚糖3(GPC3)增强型绿色荧光蛋白真核表达载体,研究GPC3对人肝癌细胞株MHCC97-L增殖的影响。方法用基因重组及限制性内切酶技术构建并鉴定表达载体pEGFP-c3-GPC3,通过脂质体介导转染人肝癌细胞株MHCC-97L,流式细胞术和G418筛选建立稳定细胞株;采用荧光定量PCR、Western blotting分别检测MHCC-97L中GPC3 mRNA和蛋白的表达情况,并在荧光显微镜下观察细胞荧光;MTT法检测GPC3对肝癌细胞MHCC97-L体外增殖能力的影响。结果经酶切及测序鉴定,确认成功构建真核表达载体pEGFP-c3-GPC3,荧光显微镜下观察到绿色荧光表达;从mRNA和蛋白水平证实,GPC3基因在转染后的MHCC-97L中成功表达;MTT生长曲线证实MHCC97-L/GPC3较其他两株细胞增殖显著加强(P<0.001)。结论成功构建pEGFR-c3-GPC3真核表达载体;筛选出稳定表达GPC3基因的MHCC97-L/GPC3细胞株;GPC3基因表达升高可促进肝癌细胞株MHCC97-L的增殖。
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| 关 键 词： | 磷脂酰肌醇蛋白聚糖3 绿色荧光蛋白 MHCC-97L 真核表达载体/构建 肝癌 |
Effect of glypican-3 on the proliferation of human hepatoma cell line MHCC97-L in vitro |
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| QIN Ru-zhai,LIU Fei-ye,CHEN Bin,XIE Jian-ming,YANG Yang,ZHENG Da-yong,LUO Rong-cheng.Effect of glypican-3 on the proliferation of human hepatoma cell line MHCC97-L in vitro[J].Journal of Southern Medical University,2011,31(3):448-452. |
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| Authors: | QIN Ru-zhai LIU Fei-ye CHEN Bin XIE Jian-ming YANG Yang ZHENG Da-yong LUO Rong-cheng |
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| Institution: | QIN Ru-zhai,LIU Fei-ye,CHEN Bin,XIE Jian-ming,YANG Yang,ZHEGN Da-yong,LUO Rong-cheng Oncology Center,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China |
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| Abstract: | Objective To construct glypican-3(GPC3)-green fluorescent protein eukaryotic expression vector pEGFP-c3-GPC3,and analyze the effect of GPC3 on the proliferation of human hepatoma cell line MHCC-97L.Methods The eukaryotic expression vector pEGFP-c3-GPC3 was constructed with recombinant DNA technique and transfected into MHCC-97L cells via Lipofectamine 2000.The cells stably expressing GPC3 were screened by flow cytometry and G418.The mRNA expression of GPC3 was detected by RT-QPCR method,and the protein expr... |
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| Keywords: | MHCC-97L |
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