| 摘要: |
| 目的:观察转化生长因子β1(TGFβ1)转入对人肝癌细胞系癌基因表达的影响。方法:pL（TGFβ1）SN和相应重组病毒上清用脂质体转染包装细胞及肝癌细胞，经G418筛选后，进行免疫组化染色，观察癌基因蛋白表达的改变。结果:提取的TGFβ1cDNA连接物经酶切鉴定，证明有正向插入及反向插入两类。病毒上清TGFβ1含量在16-41ng/ml,套式PCR后均出现290bp条带。转基因后肝癌细胞癌基因蛋白免疫组化染色结果:c-myc，c-met及ras癌基因表达，反向TGFβ1转入者为(+)？(++)，而正向转入均为(-)。免疫组化检测其TGFβ1表达蛋白，空白对照染色(±)，正向则为(++)。结论:正向插入的TGFβ1cDNA可使癌基因c-myc，c-met及ras表达减少，提示它在体外对肝癌细胞具有负调控作用。 |
| 关键词: 转染生长因子β1、肝肿瘤、癌基因 |
| DOI： |
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| In vitro observation of the oncogene expression of human hepatocellular carc inoma cells by TGFβ1 gene transfer |
| 孔宪涛,高峰,张晓琴,曹广文,封岩 |
| () |
| Abstract: |
| Objective: To investigate the effect of transforming growth fact or β1 gene transfer on the oncogene expression. Methods: Positi ve and reverse pL(TGFβ1 )SN were used to transfect PA317 and Ψ2 packaging cel l line by lipofectamine. After G418 selection, the positive cloning and serum of retroviral were obtained. Meanwhile, retroviral serum and positive, reverse TGF β1 gene were used for transfecting hepatocellular carcinoma cell line. Pos itive cell slide for immunohistochemical staining was prepared to observe change s in oncogene expression. Results: PL(TGFβ1 )SN was digested b y restriction enzyme. Positive and reverse fragments were formed. 290 bp Neo gene band was discovered by nested primer RT-PCR in all the samples. Conc lusion: Positive TGFβ1 gene reduces the c-myc, c-met and ras gene exp ression suggesting that TGFβ1 could act as a negative growth regulator for hu man hepatocellular carcinoma cells in vitro. |
| Key words: transforming growth factor beta liver neoplasms oncogenes |
