Wang Yongzhou, Xia Jiyi, Tang Xiaoping, Tang Li, Mao Xiguang, Zhang Yujiao, Yu Xiaolan
Department of Obstetrics and Gynecology, Affiliated TCM Hospital of Southwest Medical University, Luzhou 646000, China.
Experimental Medicine Center, Affiliated Hospital of Southwest Medical University, Department of Science and Technology, Southwest Medical University, Luzhou 646000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Nov;32(11):1507-1512.
Objective To investigate the effects of baicalein and U0126 treatment on the apoptosis of human cervical carcinoma HeLa cells and the potential mechanism. Methods HeLa cells were subjected to (1, 2, 5, 10, 20, 50, 100, 200, 300) μmol/L baicalein or (1, 2, 5, 10, 20, 30) μmol/L U0126 treatment for 24 hours. The optimal concentrations of baicalein and U0126 for HeLa inhibition was determined by a cell counting Kit-8 assay. HeLa cells were then treated with these inhibitory concentrations for 24 hours separately or in combination. The cell cycle and the degree of apoptosis were analyzed by flow cytometry. The cell apoptosis index was evaluated by the TUNEL assay. The expressions of extracellular signal-regulated kinase 1/2 (ERK1/2), Bax, and Bcl-2 at the mRNA and protein levels were examined by real-time PCR and Western blotting, respectively. Results Optimal inhibitory concentrations of baicalein and U0126 for HeLa cells were 200 μmol/L and 10 μmol/L, respectively. Compared with the control group, baicalein treatment increased the growth rate of cells in the G0/G1 phase but decreased the S phase. Combination treatment of 200 μmol/L baicalein and 10 μmol/L U0126 for 24 hours further reduced the S phase growth rate. Treatment with 10 μmol/L U0126 or 200 μmol/L baicalein for 24 hours induced cell apoptosis, and the combination treatment induced more apoptosis. Treatment by baicalein alone or in combination with U0126 for 24 hours significantly decreased ERK1/2 and Bcl-2 mRNA expressions, and upregulated Bax mRNA expression. It also downregulated ERK1/2 phosphorylation and Bcl-2 protein expression, while increasing Bax protein expression. Conclusion Both baicalein and U012 appear to inhibit proliferation, induce apoptosis, and increase the growth rate in the G0/G1 phase but reduce the S phase of HeLa cells. This effect is enhanced when they are used synergistically.
目的 探讨黄芩苷和U0126处理对人宫颈癌HeLa细胞凋亡的影响及其潜在机制。方法 将HeLa细胞分别用(1、2、5、10、20、50、100、200、300)μmol/L黄芩苷或(1、2、5、10、20、30)μmol/L U0126处理24小时。通过细胞计数试剂盒-8法确定黄芩苷和U0126对HeLa细胞抑制的最佳浓度。然后将HeLa细胞分别用这些抑制浓度单独或联合处理24小时。通过流式细胞术分析细胞周期和凋亡程度。通过TUNEL法评估细胞凋亡指数。分别通过实时PCR和蛋白质印迹法检测细胞外信号调节激酶1/2(ERK1/2)、Bax和Bcl-2在mRNA和蛋白质水平的表达。结果 黄芩苷和U0126对HeLa细胞的最佳抑制浓度分别为200μmol/L和10μmol/L。与对照组相比，黄芩苷处理增加了G0/G1期细胞的生长速率，但降低了S期细胞的生长速率。200μmol/L黄芩苷和10μmol/L U0126联合处理24小时进一步降低了S期细胞的生长速率。10μmol/L U0126或200μmol/L黄芩苷处理24小时诱导细胞凋亡，联合处理诱导更多凋亡。黄芩苷单独或与U0126联合处理24小时显著降低ERK1/2和Bcl-2 mRNA表达，并上调Bax mRNA表达。它还下调ERK1/2磷酸化和Bcl-2蛋白表达，同时增加Bax蛋白表达。结论 黄芩苷和U0126似乎均能抑制HeLa细胞增殖、诱导凋亡，并增加G0/G1期细胞生长速率，但降低S期细胞生长速率。二者协同使用时，这种作用增强。
