| 罗春华,朱建思.稳定表达外源性p16基因肝癌SMMC-7721细胞株的建立及鉴定.[J].中南医学科学杂志.,2005,(2):184-187. |
| 稳定表达外源性p16基因肝癌SMMC-7721细胞株的建立及鉴定 |
| Establishment and Identification of SMMC-7721 Cell Line With Stable Expression of Exogenous p16 Gene |
| 修订日期：2004-12-10 |
| DOI： |
| 中文关键词: p16抑癌基因 基因转染 肝肿瘤 |
| 英文关键词:p16,gene transfection,hepatocellular carcinoma |
| 基金项目:湖南省卫生厅课题基金 (4 -0 2 -WS -YO2 0 68) |
| 罗春华 朱建思 |
| 南华大学肿瘤研究所,湖南衡阳421001 |
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| 中文摘要: |
| 目的构建稳定表达外源性p16基因的肝癌SMMC-7721细胞株。方法利用脂质体介导的基因转染方法，借助真核质粒表达载体pcDNA3．1( )，将外源性p16基因转染入此基因表达下调的人肝癌细胞株SMMC-7721细胞中，经G418筛选，建立稳定表达的细胞株，用逆转录聚合酶链反应(RT—PCR)及免疫组织化学法鉴定p16基因的表达，同时对细胞株分泌蛋白进行活性检测。结果转染p16基因的SMMC-7721细胞中可以检测到p16mRNA及蛋白的表达。结论建立稳定表达P16抑癌蛋白的SMMC-7721细胞株有助于研究p16抑癌基因在肝癌发生中的作用。 |
| 英文摘要: |
| Objective To establish SMMC-7721 cell line with stably expressing tumor suppressor gene p16. Methods The exogenous tumor suppressor gene p16 was transfected into SMMC-7721 cells with lipofectamine TM 2000 by eukaryotic expressive vector(pcDNA3.1 (+) ) . We got clone cells after selected by G418. All the clone cells were detected by RT- PCR and immunocytochemistry staining on mRNA and protein levels. Meanwhile,P16 protein activity were measured by MTT. Results p16 mRNA and protein were expressed in those transfected with p16 gene cells. Conclusion Exogenous p16 gene may stably expressed in human hepatocellular carcinoma cell line SMMC-7721. The establishment of the cell line can help to study the functions of tumor suppression gene p16 in hepatocellular carcinoma cells. |
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