索拉非尼对Hela细胞增殖及VEGFR-3和PDGFR-β表达的影响

摘要

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.