摘要
目的探讨Hedgehog(Hh)信号在SMMC-7721肝癌细胞运动侵袭能力中的作用及机制。方法应用KAAD-环巴胺(KAAD-cyclopamine)特异性阻断Hh信号,通过细胞划痕愈合实验以及Transwell小室检测肝癌细胞的迁移速率与侵袭能力,蛋白质印迹法检测E-钙黏蛋白(E-cad)在肝癌细胞中的表达差异。结果 Hh信号通路失活后,肝癌细胞迁移率由(41.3±7.0)μm/h显著下降至(24.9±3.8)μm/h(P<0.01),运动及侵袭能力明显减弱,穿过Transwell小室和人工基底膜的肝癌细胞数量分别减少了(54.4±7.0)%和(48.3±11.9)%(P<0.05)。随着Hh信号下调,E-cad在肝癌细胞中的表达量较前升高了2.32倍(P<0.01)。结论 Hh信号通路与肝癌细胞侵袭力密切相关,KAADcyclopamine抑制Hh信号通路可导致SMMC-7721细胞E-cad的表达上调,抑制肝癌细胞侵袭力。
Objective To explore the role of Hedgehog（Hh）signaling pathway in the migration and invasion of hepatocarcinoma SMMC-7721 cells and its mechanism.Methods Hedgehog signaling pathway was specifically blocked by KAAD-cyclopamine.Wound healing assay and Transwell assay were employed to detect the migration rate and invasiveness of SMMC-7721 cells and Western blot to measure the expression of E-cad in these cells.Results After inhibition of Hedgehog signaling pathway,the migration rate of SMMC-7721 cells was significantly decreased from（41.3±7.0）μm/h to（24.9±3.8）μm/h（P0.01）,the migrating and invasive capacity of these cells was conspicuously reduced,and the number of HCCs crossing the Transwell and matrigel was reduced by（54.4±7.0）% and（48.3±11.9）%（P0.05）,respectively.The expression of E-cad protein in SMMC-7721 cells after blockage of Hedgehog signaling pathway was 2.32 times higher than that before inhibition（P0.01）.Conclusion Hh signaling pathway is closely related to the invasiveness of hepatocarcinoma cells.KAAD-cyclopamine may suppress the invasiveness of hepatocarcinoma cells by inhibiting the Hh signaling pathway and up-regulating the expression of E-cad.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2015年第2期143-146,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81202121)
武汉市卫生局科研项目(No.WX12C07)
