黄芩素联合U0126促进人膀胱癌T24细胞凋亡的分子机制研究

摘要

目的 探讨黄芩素联合U0126促进人膀胱癌细胞系T24凋亡的分子机制.方法 用黄芩素和U0126处理人膀胱癌T24细胞株,(1)CCK8检测细胞增殖;(2)流式细胞仪检测T24细胞周期的变化;(3)显微镜观察细胞凋亡;(4)流式细胞仪检测早期细胞凋亡和晚期细胞凋亡;(5)Real Time PCR检测细胞增殖和凋亡相关蛋白的表达水平;(6)Western blot检测细胞增殖和凋亡相关蛋白的表达水平.结果 黄芩素或U0126浓度越高,T24细胞增殖能力逐渐下降(P<0.05);不同浓度的黄芩素刺激人膀胱癌细胞株T24,T24细胞数量随浓度上升而逐渐减少(P<0.05);黄芩素与U0126联合处理组的T24细胞株S期的细胞数量显著低于黄芩素、U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞株早期凋亡和晚期凋亡均显著高于黄芩素、U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞株ERK1/2、Bcl-2和P38的mRNA表达水平显著低于黄芩素或U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞ERK1/2、P38的磷酸化水平以及Bcl-2的蛋白表达量均显著低于黄芩素或U0126单独处理组(P<0.05).结论 黄芩素和U0126均可显著抑制人膀胱癌细胞增殖,诱导T24早期凋亡和晚期凋亡且具有浓度依赖性,且两者具有协同效应.因此,黄芩素和U0126可用于治疗膀胱癌.%Objective To investigate the molecular mechanism of baicalein combined with U0126 in promot-ing apoptosis of human bladder cancer cell line T24. Methods Human lung cancer T24 cell line was treated with baica-lein and U0126. CCK8 was used to detect cell proliferation, and flow cytometry was used to detect the cell cycle of T24 cells. Cell apoptosis was observed by microscope, and early cell apoptosis and late cell apoptosis were detected by flow cytometry. Cell proliferation and apoptosis-related protein expression levels were detected by Real time PCR and west-ern blot (P<0.05). Results The proliferation ability of human bladder cancer cell lines T24 decreased with the increase of concentration of baicalein or U0126 (P<0.05). When treated with baicalein, the number of T24 cells decreased with the increase of the concentration of baicalein (P<0.05). The number of cells in S phase of T24 cell line treated with baica-lein and U0126 was significantly lower than that of baicalein and U0126 alone (P<0.05), and the early apoptosis and late apoptosis of T24 cell line treated with baicalein and U0126 were significantly higher (P<0.05). The expression of ERK1/2, Bcl-2 and P38 mRNA in the T24 cell line treated with baicalein and U0126 was significantly lower than that of baicalein or U0126 (P<0.05). The phosphorylation level of ERK1/2 and P38 and the protein expression of Bcl-2 in T24 cells treated with baicalein and U0126 were significantly lower than those of bauxine or U0126 alone (P<0.05). Conclusion Both baicalein and U0126 can significantly inhibit the proliferation of human bladder cancer cells, induce early apoptosis and late apoptosis of T24, in a synergistic effect. Therefore, baicalein and U0126 can be used to treat bladder cancer.