【摘要】 目的:探讨汉黄芩素对人结肠癌细胞HCT116和HT29细胞系的增殖、侵袭能力的影响,并研究汉黄芩素对酪氨酸3/色氨酸5-单加氧酶激活蛋白(Ywhaz)蛋白水平的影响。方法:体外培养HCT116和HT29细胞系,设空白组、汉黄芩素不同浓度组(浓度分别为5,10,20,40μmol·L-1)作用不同时间后,采用细胞计数试剂盒(CCK-8)法检测汉黄芩素对结肠癌细胞增殖的影响,并用Annexin V-FITC/PI双标后流式细胞仪检测结肠癌细胞的凋亡率;穿透小室(Transwell)小室法检测处理24 h后细胞侵袭和迁移力的变化;实时荧光定量-聚合酶链式反应(q PCR)检测不同浓度汉黄芩素作用24 h后Ywhaz mRNA的水平,并运用蛋白质免疫印迹(Western blot)检测该蛋白及其磷酸化水平。结果:与空白组比较,汉黄芩素可明显抑制结肠癌细胞系的增殖,具有浓度和时间依赖性,并促进结肠癌细胞系的凋亡率,其中20和40μmol·L-1的汉黄芩素抑制细胞增殖作用显著(P<0.01),且不同浓度的汉黄芩素(10,20,40μmol·L-1)作用于结肠癌细胞后,明显降低肿瘤细胞的穿膜数(P<0.05,P<0.01),汉黄芩素可下调Ywhaz mRNA水平和蛋白水平的表达,并降低Ywhaz的磷酸化水平(P<0.05,P<0.01)。结论:汉黄芩素可显著抑制HCT116和HT29细胞系的增殖、侵袭和迁移能力,并诱导细胞凋亡,其抗肿瘤机制可能与下调Ywhaz的蛋白水平及其蛋白的磷酸化水平有关。更多还原

【Abstract】 Objective: To study the effect of wogonin on inhibiting the proliferation,migration and invasion of human colorectal cancer cells（ HCT116 and HT29 cell lines）,and investigate its effect on tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein（ Ywhaz） expression levels. Method: HCT116 and HT29 cell lines were cultured in vitro,and then treated with wogonin of different concentrations( 5,10,20,40μmol·L^-1) for various periods. Another blank group was also set up. Then the cell counting kit（ CCK-8） was used to detect the effect of wogonin on proliferation of colorectal cancer cells; Annexin V-FITC / PI double standard flow cytometry was used to detect the apoptosis of colorectal cancer cells; Transwell method was used to detect the cells migration and invasioncapacities after 24 h treatment; Fluorescense real-time quantitative PCR method was used to detect mRNA expression levels of Ywhaz after 24 h treatment with different concentrations of wogonin,and their protein expression level and phosphorylation level were detected by Western blot assay. Result: As compared with blank group, wogonin significantly inhibited the proliferation of colorectal cancer cell lines in a dosedependent manner, and promoted apoptosis of colorectal cancer cell lines; the inhibitory effect on cells proliferation was significant by 20,40 μmol·L^-1 wogonin（ P < 0. 01）. Moreover, different concentrations of wogonin( 10,20,40 μmol·L^-1) could significantly reduce the number of transmembrane tumor cells（ P < 0. 05,P < 0. 01）,down-regulate the protein and mRNA expression levels of Ywhaz and reduce the phosphorylation level of Ywhaz in HCT116 and HT29 cell lines（ P < 0. 05,P < 0. 01）. Conclusion: Wogonin can significantly inhibit the proliferation,migration and invasion of HCT116 and HT29 cell lines and induce apoptosis. Its anti-tumor mechanism may be associated with down-regulating the protein expression of Ywhaz and its phosphorylation level.更多还原