羟基红花黄色素A在结直肠癌发生发展中的影响及其机制

摘要

目的 观察羟基红花黄色素A(HSYA)对结直肠癌细胞增殖、侵袭、迁移和凋亡的影响及其分子机制.方法 采用HSYA低、中、高剂量作用于HT-29细胞,在24 h、48 h、72 h用MTT法检测细胞抑制率,Transwell实验分析HT-29细胞迁移和侵袭情况,流式细胞术分析HSYA对HT-29细胞凋亡及周期分布的影响;免疫印迹法(western blot)检测E-cad、Vi、FN、TGF-β、Smad2和α-SMA蛋白的表达水平.结果 HSYA作用于HT-29细胞后.HT-29细胞体外增殖均受到抑制(均P＜0.05),其抑制细胞增殖的效果与HSYA的浓度和作用时间有关.HSYA明显抑制HT-29细胞的侵袭和转移(均P＜ 0.05),同时HT-29细胞凋亡率也均增高(均P＜ 0.05),表现为G0/G1期细胞比例增加(均P＜ 0.05),S期细胞比例降低(均P＜ 0.05).HSYA能够通过增加E-cad蛋白表达(均P＜ 0.05)和减少Vi和FN蛋白表达(均P＜0.05)抑制HT-29细胞上皮间质化(EMT)过程的影响.随着HSYA的浓度升高,TGF-β 、Smad2和α-SMA蛋白表达均下降(均P＜ 0.05).结论 HSYA能够抑制结直肠癌细胞增殖、侵袭、迁移、EMT和促进结直肠癌细胞凋亡,其机制可能与抑制TGF-β信号通路激活有关,HSYA可能成为结直肠癌的治疗靶点.%Objective:To study the effects of hydroxysafflor yellow A (HSYA) on the proliferation,invasion,migration and apoptosis in colorectal cancer cell and its mechanism.Methods:The low,middle and high dose HSYA were applied to treat HT-29 cells.The MTT assay was used to observe the inhibiting effects of HSYA on the cell proliferation at 24 h,48 h and 72 h.Transwell experiment was used to investigate the migration and invasion in HT-29 cells.The flow cytometry (FCM) was adopted to investigate the influence of HSYA on the HT-29 cells apoptosis and cell cycle.Western blot was used to measure the expression level of E-cad,Vi,FN,TGF-β,Smad2 and α-SMA in cells.Results:After HT-29 cells were disposed by HSYA,significantly inhibiting effects on the proliferation of HT-29 cell were observed(all P＜0.05).The effect was correlated with the density and action time of HSYA.HSYA could significantly inhibit the migration and invasion in HT-29 cells (all P＜ 0.05),induce the apoptosis of HT-29 cell(all P＜0.05) and change cell cycle which showed mainly the percentage of HT-29 cell increased in stage G0/G1 (all P＜0.05) and decreased in stage S (all P＜0.05).HSYA promoted E-cad protein expression (all P＜0.05) and decreased protein expression of Vi and FN (all P＜ 0.05) to inhibit epithelial to mesenchymal transition in HT-29 cells.Moreover,with the increase of HSYA concentrations,it significantly increased the protein expression level of TGF-β,Smad2 and α-SMA (all P＜ 0.05).Conclusions:HSYA can inhibit the proliferation,invasion,migration and EMT of colorectal cancer cell,and promote the apoptosis of colorectal cancer cells.The mechanism may be related to the inhibition of TGF-β signaling pathway activation,and HSYA may be a therapeutic target for colorectal cancer.