羽扇豆醇通过抑制RhoA-ROCK1信号通路抑制结肠癌细胞增殖

该文探讨了羽扇豆醇(Lupeol)对人结肠癌HCT116和SW620细胞增殖的影响及相关作用机制。使用不同浓度的Lupeol处理HCT116和SW620细胞后,用MTT法检测细胞活性,CCK8法检测细胞增殖能力,平板克隆实验检测细胞克隆形成能力,流式细胞术检测细胞周期和细胞凋亡,(quantitative real-time PCR,qPCR)和Western blot检测相应mRNA和蛋白表达水平,免疫荧光检测β-Catenin蛋白细胞内分布情况。通过构建shRNA敲低两种结肠癌细胞中RhoA,进一步研究Lupeol影响细胞增殖的分子机制。结果显示,Lupeol处理后,HCT116和SW620细胞增殖能力明显下降,克隆形成能力受到抑制,细胞周期阻滞于G0/G1期,细胞内RhoA、ROCK1、β-Catenin、Cyclin D1 mRNA和蛋白表达水平均显著下降,β-Catenin蛋白胞质和胞膜上分布减少。敲低RhoA后抑制了细胞增殖,同时使得RhoA-ROCK1-β-Catenin信号通路蛋白受到抑制,β-Catenin蛋白胞质和胞膜上分布减少。综上所述,Lupeol可通过抑制RhoA-ROCK1信号通路,抑制β-Catenin蛋白表达,进而抑制HCT116和SW620细胞增殖,Lupeol有望成为临床结肠癌治疗的新药物。

Lupeol Inhibits Proliferation of Colorectal Cancer Cells by Suppressing RhoA-ROCK1 Signaling Pathway

This study aimed to explore the effect of Lupeol on the proliferation of CRC(colorectal cancer)cells and its underlying mechanism.HCT116 and SW620 cells were treated with Lupeol for 48 h.Then,MTT assay,CCK8 assay,plate clone assay,and flow cytometry were performed to detect the cell viability,proliferation ability,colony-forming ability,cell cycle,and cell apoptosis.The qPCR(quantitative real-time PCR)and Western blot experiments were used to evaluate the mRNA and protein expression levels.Immunofluorescence assay was used to explore the intracellular distribution of the β-Catenin protein.Further,RhoA was knockdown in CRC cells to explore the mechanism of Lupeol.Our results showed that Lupeol inhibited the proliferation of HCT116 and SW620 cells but did not induce cell apoptosis at selected doses.And the cell cycle was arrested in the G0/G1 phase after Lupeol treatment.In addition,Lupeol downregulated the expression of RhoA,ROCK1,β-Catenin and Cyclin D1 in both CRC cells.The distribution of β-Catenin protein in the cytoplasm and membrane was reduced after Lupeol administration.Moreover,the proliferation ability was inhibited after RhoA knockdown.In conclusion,Lupeol could suppress the proliferation of HCT116 and SW620 cells by inhibiting the RhoA-ROCK1 signaling pathway and might provide an effective agent for CRC patients.