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.