RhoGAP结构域在DLC-1基因抑制人结肠癌HT29细胞增殖、侵袭中的作用

DLC-1 gene inhibits proliferation and invasion of colon cancer HT29 cells via its RhoGAP domain

目的:探讨Rho蛋白GTP酶活化蛋白(Rho GTPase activation protein,RhoGAP)结构域在肝癌缺失基因1(frequentlydeleted in liver,DLC-1)抑制人结肠癌HT29细胞的增殖、侵袭中的作用。方法:脂质体转染DLC-1基因及RhoGAP结构域缺失的ΔDLC-1亚克隆至大肠癌HT29细胞株;应用MTT、平板克隆实验检测细胞增殖的改变;Transwell实验观察细胞侵袭迁移能力的改变;流式细胞术检测细胞周期。结果:转染成功后,全长DLC-1基因转染组(pcDNA3.1-DLC1-HT29)与野生型及空载组相比,细胞增殖能力下降,侵袭能力减弱,细胞周期G1期阻滞并诱导凋亡,而ΔDLC-1亚克隆转染组(pcDNA3.1-ΔDLC1-HT29)对细胞增殖、侵袭及细胞周期均无明显影响。结论:DLC-1基因可能是通过其内在的RhoGAP结构域抑制人结肠癌细胞HT29的增殖和侵袭。

Objective：To analyze the role of Rho GTPase activating protein （RhoGAP） domain of frequently deleted in liver （DLC-1） gene in suppression of proliferation and invasion in colon cancer HT29 cells.Methods：Recombinant pcDNA3.1 vector containing the DLC-1 gene and RhoGAP domain deleted ΔDLC1 subclone was constructed and transfected into HT29 cell line.The cell growth was measured by MTT and colony formation assay.The ability of cell migration of HT29 cells was examined by using Transwell migration assays.Furthermore,cell cycle was evaluated by flow cytometry.Results：After transfected successfully,pcDNA3.1DLC1-HT29 cells showed a remarkable suppression of proliferation and invasion,as well as G1 sphase arrested and apoptosis induced,compared with wild or mock-vector HT29 cells.However,pcDNA3.1-ΔDLC1 subclone transfection had no effect on cell proliferation,invasion and cell cycle distribution.Conclusion：DLC-1 gene inhibits proliferation and invasion of colon cancer HT29 cells via its RhoGAP domain.