肝癌MHCC97H细胞肝细胞生长因子(HGF)/c-Met通路激活对血管内皮生长因子表达的影响  

The Effection of HGF/cMet Pathway on the Expression of VEGF in MHCC97H

作　　者：潘志刚[1] 刘康达[2] 胡美玉[2] 吴伟忠[3] 

机构地区：[1]复旦大学附属中山医院全科医学科,上海200032 [2]复旦大学附属中山医院实验研究中心,上海200032 [3]复旦大学附属中山医院肝癌研究所,上海200032

出　　处：《中国临床医学》2008年第5期629-632,共4页Chinese Journal of Clinical Medicine

摘　　要：目的:研究肝癌MHCC97H细胞中肝细胞生长因子(HGF)诱导血管内皮生长因子(VEGF)表达过程中的转录因子Sp1的活性。方法:将MHCC97H细胞分为对照组、HGF组、光辉霉素组,分别用RT-PCR及Western blot分析与VEGF mRNA、蛋白质表达及Sp1水平、磷酸化Sp1水平变化以及应用Sp1阻断剂光辉霉素后上述目标蛋白的变化情况。结果:外源性HGF60ng·mL-1孵育15min、16h后分别可以使MHCC97H细胞的VEGF mRNA及其蛋白质的表达明显提高(P<0.01)。这一过程中Sp1水平无明显变化(P>0.05)。进一步分析发现磷酸化Sp1水平明显升高(P<0.01)。用Sp1抑制剂光辉霉素预先处理MHCC97H细胞1h后,再加入HGF 60ng·mL-1 VEGFmRNA及其随后蛋白质表达均受到抑制(P<0.01)。结论:HGF诱导MHCC97H肝癌细胞VEGF表达是通过增强Sp1磷酸化实现的。Objective：To study the involvement of Spl in the process of HGF-induced expression of VEGF in a hepatoma cell line with high metastasis potential, MHCC97H cell line. Methods： M HCC97H cell line were cultured and divided into 3 groups： control group that without any additional stimuli; HGF group that added HGF at 60ng·mL^-1; Mithramycin group that added Mithramyein at 150 nM 1 h in advance and then added HGF at 60ng·mL^-1. RT PCR,Western blot and immunoprecipitation were applied to analysis the expression of VEGF mRNA, VEGF protein, Spl and phosphorylated Spl, respectively. Results： In HGF group, there were obviously increasing level of VEGF mRNA when HGF was added for 15min and VEGF protein elevating after 16 h later, P〈0.01 respectively. Western blot analysis showed Spl was stable in HGF group comparing with the control group （P〉0.05）, a remarkable rising of phosphorylated Spl was detected between the two groups, P〈0.01. The elevation of VEGF mRNA and protein could be notably inhibited by pretreating with Mithramyein, an inhibitor of Spl, at 150nM for lh, P〈0.01. Conclusion： HGF could induce expression of VEGF mRNA and protein in MHCC97H cell line. Spl phosphorylation play an important role in this induction.

关 键 词：肝癌 肝细胞生长因子 血管内皮生长因子 转录因子 

分 类 号：R735.7[医药卫生—肿瘤]