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目的探讨汉黄芩素对人结肠癌LoVo细胞体外增殖、凋亡和周期及G_1/S关卡调控因子的影响,为其临床应用提供理论依据。方法汉黄芩素处理LoVo细胞后,采用CCK-8法检测细胞增殖情况,Graphpad计算IC50;FCM检测细胞周期和凋亡率;Western blot检测G_1/S关卡调控因子蛋白表达。结果与对照组相比,汉黄芩素对LoVo细胞增殖抑制作用呈现时间和剂量依赖性(P<0.05);24、48和72 h的IC_(50)分别是198.7、65.1和38.6μmol·L~(-1)。不同浓度汉黄芩素作用48 h后,汉黄芩素组LoVo细胞凋亡率均明显高于对照组,且呈现出剂量依赖性(P<0.05);LoVo细胞G_0/G_1期比例增大(P<0.05),S期比例减少(P<0.05),G2/M期比例变化不明显(P>0.05);随着汉黄芩素浓度增加,Cyclin D1、Cyclin E、CDK2和CDK4蛋白相对表达量均逐渐下调(P<0.05),而p21和p27蛋白相对表达量则逐渐上调(P<0.05)。Spearman相关分析显示,LoVo细胞G_0/G_1期比例与p21和p27蛋白表达呈正相关,而与Cyclin D1、Cyclin E、CDK2和CDK4蛋白表达呈负相关。结论汉黄芩素通过G_1/S关卡调控因子蛋白表达来阻滞结肠癌LoVo细胞于G_0/G_1期,进而有效地抑制细胞增殖和诱导细胞凋亡,提示汉黄芩素在结肠癌的辅助治疗中有一定的应用前景。
Objective To investigate the effect of wogonin on proliferation, apoptosis, cycle and G_1 / S level regulatory factors of human colon cancer LoVo cells in vitro and to provide a theoretical basis for its clinical application. Methods The proliferation of LoVo cells was detected by the method of CCK-8. The IC50 was determined by Graphpad. The cell cycle and apoptosis rate were detected by FCM. The protein expression of G_1 / S level regulator was detected by Western blot. Results Compared with the control group, the inhibitory effect of wogonin on LoVo cells was time-and dose-dependent (P <0.05). The IC 50 values at 24, 48 and 72 h were 198.7, 65.1 and 38.6 μmol·L- (-1). The apoptosis rate of LoVo cells was significantly higher than that of the control group at different concentrations of wogonin for 48 h (P <0.05), and the proportion of G0 / G1 phase of LoVo cells increased (P < 0.05). The proportion of S phase was decreased (P <0.05) and the ratio of G2 / M phase was not changed significantly (P> 0.05). The relative expression of Cyclin D1, Cyclin E, CDK2 and CDK4 protein gradually increased with the increase of wogonin (P <0.05), while the relative expression of p21 and p27 protein gradually increased (P <0.05). Spearman correlation analysis showed that the proportion of G0 / G1 phase of LoVo cells was positively correlated with the expression of p21 and p27, but negatively correlated with the expressions of Cyclin D1, Cyclin E, CDK2 and CDK4. Conclusion Wogonin can block LoVo cells in G_0 / G_1 phase through the expression of G_1 / S regulators, which can effectively inhibit cell proliferation and induce apoptosis. It suggests that Wogonin has the potential of adjuvant treatment of colon cancer Certain application prospects.