论文部分内容阅读
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?
Objective To study the effect of hepatocyte growth factor (HGF) on the apoptosis and expression of bcl-2 gene in hepatocellular carcinoma cells induced by VP16. Methods We treated SMMC-7721 cells with VP-16 combined with HGF. Hepatoma cells were divided into three groups: VP 16 (0 5, 1, 5 μM/L); HGF (10, 20, 30 μg/L) + VP 16 (0 5 μM/L); saline control group. Western blot analysis of bcl 2 and nuclear proliferating antigen (PCNA) expression, DNA gel electrophoresis, inverted microscope and HE staining observed cell morphology. Results VP 165μM/L, 05μM/L, HGF+VP16, apoptosis occurred after 5h, 7d and 72h respectively. The apoptosis was not obvious with 05μM/LVP16 alone. DNA electrophoresis: 05～1μM/LVP16 treatment for 1h had no effect on DNA. At 24h, DNA was formed around 180bp at 1μM/L and 5μM/L. After 36h, HGF+VP-1605μM/L group was also concentrated at 180bp. Western blot: SMMC 7721 cells expressed PCNA to varying degrees, and VP 16+HGF had the highest expression. Bcl 2 in the 5μM / L group 24h, 1μM / L group 72h, HGF + VP 16 (0 5μM/L) 96h decreased expression, 120h after no expression. Conclusion HGF promotes apoptosis of hepatoma cells induced by VP16.