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

Effects of Hydroxysafflor Yellow A on the Development and Progression in Colorectal Cancer and Its Mechanism

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 hydroxysaffior yellow A （HSYA） on the proliferation,invasion,mi- gration 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 MTr 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 inva- sion in HT-29 ceils. 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 ac- tion time of HSYA. HSYA could significantly inhibit the migration and invasion in HT-29 cells（all P〈 0.05）,in- duce the apoptosis of HT-29 ceil（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 in- creased the protein expression level of TGF-β,Smad2 and α-SMA （all P〈 0.05）. Conclusions： HSYA can in- hibit the proliferation, invasion, migration and EMT of colorectal cancer cell, and promote the apoptosis of colorec- tal cancer cells. The mechanism may be related to the inhibition of TGF-β signnling pathway activation,and HSYA may be a therapoutic target for colorectal cancer.