摘要
目的研究肝细胞生长因子（ＨＧＦ）对ＶＰ１６诱导肝癌细胞凋亡及ｂｃｌ－２基因表达的影响．方法我们用ＶＰ１６联合ＨＧＦ处理肝癌ＳＭＭＣ７７２１细胞．肝癌细胞处理分３组：ＶＰ１６（０５，１，５μＭ／Ｌ）；ＨＧＦ（１０，２０，３０μｇ／Ｌ）＋ＶＰ１６（０５μＭ／Ｌ）；生理盐水对照组．Ｗｅｓｔｅｒｎｂｌｏｔ分析ｂｃｌ２及核增殖抗原（ＰＣＮＡ）表达，ＤＮＡ凝胶电泳，倒置显微镜及ＨＥ染色观察细胞形态．结果应用ＶＰ１６５μＭ／Ｌ，０５μＭ／Ｌ，ＨＧＦ＋ＶＰ１６，细胞凋亡分别在５ｈ，７ｄ和７２ｈ后发生，单用０５μＭ／ＬＶＰ１６诱发凋亡不明显．ＤＮＡ电泳∶０５～１μＭ／ＬＶＰ１６处理１ｈ对ＤＮＡ无影响，２４ｈ时１μＭ／Ｌ和５μＭ／Ｌ组ＤＮＡ在１８０ｂｐ左右形成条带．３６ｈ后，ＨＧＦ＋ＶＰ１６０５μＭ／Ｌ组也在１８０ｂｐ处集中．Ｗｅｓｔｅｒｎｂｌｏｔ：ＳＭＭＣ７７２１细胞均不同程度表达ＰＣＮＡ，ＶＰ１６＋ＨＧＦ组表达量最高．ｂｃｌ２在５μＭ／Ｌ组处理后２４ｈ，１μＭ／Ｌ组７２ｈ，ＨＧＦ＋ＶＰ１６（０５μＭ／Ｌ）９６ｈ表达下降，１２０ｈ后未见表达．结论ＨＧＦ促进ＶＰ１６诱发肝癌细胞凋亡?
AIM To study the hepatocyte growth factor (HGF) in etoposide (VP 16) induced apoptosis and bcl 2 gene. METHODS Apoptosis (hepatic cancer cell line, SMMC 7721) was induced by VP 16 (0 5, 1, 5μM/L), HGF (10, 20, 30μg/L)+VP 16 (0 5μM/L) and physiological salt solution served as control. The morphology of apoptosis was observed by HE staining. By Western blot, the expressions of bcl 2 and PCNA protein were analyzed in induced apoptosis. RESULTS Apotosis was found after 5h, 7 days and 72h in VP 16 5μM/L, 0 5μM/L and HGF (10, 20, 30μg/L)+VP 16 (0 5μM/L) groups, respectively. It was not killed completely by single 0 5μM/L VP 16 after 7 days. Western blot analysis of bcl 2 showed that the expressions were reduced in VP 16 5μM/L group for 24h, 1μM/L for 72h, HGF+VP 16 for 96h, respectively. No expression of the oncoprotein was seen after 120h. The expressions of PCNA were the highest in VP 16+HGF group. CONCLUSION HGF might improve etoposide induced apoptosis in hepatic carcinoma cells. The proliferating carcinoma cells may be regulated, and bcl 2 gene altered by their combination.
关键词
肝细胞生长因子
药理学
原癌基因
肝癌
细胞凋亡
Hepatocyte growth factor/pharmacology\ \ Etoposide/pharmacology\ \ Apoptosis/drug effects\ \ Liver neoplasms/drug therapy\ \ Proto_oncogenes/drug effects\ \ Gene/expression/drug effects
