索拉非尼对Hela细胞增殖及VEGFR-3和PDGFR-β表达的影响

Effect of sorafenib on proliferation of Hela cells and the expression of VEGFR-3 and PDGFR-β in Hela cells

目的:研究索拉非尼能否抑制宫颈癌Hela细胞增殖、抑制Hela细胞中VEGFR-3和PDGFR-β的表达。方法:四甲基偶氮唑蓝(MTT)快速比色法检测索拉非尼作用于Hela细胞24h、48h、72h后细胞增殖率,Hoechst荧光染色观察药物作用(1mg/L)48h后的凋亡细胞,细胞免疫荧光法和Western blot检测不同浓度的索拉非尼(1mg/L、0.1mg/L、0.01mg/L)作用Hela细胞VEGFR-3和PDGFR-β表达情况。结果:不同浓度的索拉非尼作用Hela细胞后,Hela细胞的增殖率下降,呈浓度依赖性(P<0.05),随着药物浓度的增加Hela细胞的增殖率不断下降。药物作用后能观察到凋亡的Hela细胞,细胞核变小、固缩,荧光明显增强。正常Hela细胞中有VEGFR-3和PDGFR-β表达,索拉非尼作用后,Hela细胞绿色荧光明显减弱,VEGFR-3和PDGFR-β表达明显降低,呈浓度依赖性。结论:索拉非尼能够诱导Hela细胞的凋亡,抑制其增殖且呈浓度依赖性。索拉非尼能够抑制Hela细胞VEGFR-3和PDGFR-β表达,可能降低Hela细胞侵袭转移的能力。

Objective:To investigate whether sorafenib can inhabit the proliferation of Hela cells and inhibit the expression of VEGFR-3 and PDGFR-βin Hela cells.Methods:MTT assay was used to analyze cells proliferation rate after treatment with different concentration of sorafenib for 24h,48h,72h.Hoechst 33258 was used to stain nucle-us of Hela cells treatment with sorafenib (1.0mg/L),48h later,observed nucleus morphological changes.Immunoflu-orescence and Western blot was used to observe the expression of VEGFR-3 and PDGFR-βin Hela cells with the treatment of different concentration of sorafenib (1mg/L,0.1mg/L,0.01mg/L).Results:After treatment with differ-ent concentration of sorafenib ,the proliferation rate of Hela cells was down ,and it has dose-dependent effect (P<0.05).Sorafenib can make Hela cells apoptosis.The apoptotic cells,nuclear small,pyknosis,fluorescence enhanced obviously.We can observe the expression of VEGFR-3 and PDGFR-βin Hela cells.The green fluorescence was re-duced with the treatment of sorafenib,the expression of VEGFR-3 and PDGFR-βin Hela cells were down,and it had dose -dependent effect.Conclusion:Sorafenib can make Hela cells apoptosis,inhibit the proliferation of Hela cells and it has dose-dependent effect.Sorafenib can inhibit the expression of VEGFR-3 and PDGFR-βin Hela cells,maybe sorafenib can decrease the property of its invasion and metastasis.