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。
Objective: To investigate the effect of transforming growth factor 1 gene transfer on the oncogene expression. Methods: Positive and reverse pL(TGF1 )SN were used to transfect PA317 and 2 packaging cell line by lipofectamine. After G418 selection, the positive cloning and serum of retroviral were obtained. Meanwhile, retroviral serum and positive, reverse TGF1 gene were used for transfecting hepatocellular carcinoma cell line. Positive cell slide for immunohistochemical staining was prepared to observe changes in oncogene expression. Results: PL(TGF1 )SN was digested by restriction enzyme. Positive and reverse fragments were formed. 290 bp Neo gene band was discovered by nested primer RTPCR in all the samples. Conclusion : Positive TGF1 gene reduces the cmyc, cmet and ras gene expression suggesting that TGF1 could act as a negative growth regulator for human hepatocellular carcinoma cells in vitro.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1999年第5期292-294,F003,共4页
Academic Journal of Second Military Medical University
