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目的探讨维生素C与维生素K3联合用药能否诱导人膀胱癌细胞株T24发生凋亡及机制。方法 (1)用噻唑蓝(MTT)法筛选出维生素C作用于T24细胞72h后的最佳作用浓度。(2)用该最佳浓度下的VC联合不同浓度的VK3(1,2.5,5,10μM),筛选出作用72h后的最佳联合用药浓度。(3)分组并培养各组细胞:空白对照组,最佳浓度的维生素C作用组,最佳浓度的联合用药组。(4)用碘化丙啶(PI)流式细胞术观察上述三组的细胞凋亡情况,通过流式软件FACS分析细胞周期变化。结果 (1)以MTT法筛选出单用维生素C作用72h后的最佳药物浓度为1000μM,(2)以MTT法筛选出浓度为该浓度维生素C与不同浓度的VK3联用时,VK3的最佳药物浓度为10μM。(3)流式细胞仪检测各组细胞凋亡率结果表明:10μMVK3+1000μMVC联用组与1000μMVC组相比,可显著提高细胞凋亡率(p<0.05)。(4)流式细胞仪检测各组细胞周期示:VC+K3可诱导细胞凋亡的作用集中于S期(p<0.05)。结论维生素K3与维生素C联合用药可增强诱导人膀胱癌T24细胞凋亡的作用,其机制可能与维生素C阻滞T24细胞周期于S期有关。
Objective To investigate whether vitamin C and vitamin K3 can induce the apoptosis of human bladder cancer cell line T24 and its mechanism. Methods (1) Thiazolyl blue (MTT) method was used to screen out the best effect concentration of vitamin C on T24 cells after 72 hours. (2) With the optimal concentration of VC combined with different concentrations of VK3 (1, 2.5, 5, 10 μM), the optimal combination concentration after 72 h was screened out. (3) Grouping and culture of each group of cells: blank control group, the best concentration of vitamin C group, the best concentration of combination group. (4) The apoptosis of the above three groups was observed by propidium iodide (PI) flow cytometry, and the cell cycle was analyzed by flow cytometry FACS. Results (1) The optimum concentration of vitamin C was 1000μM after single-dose vitamin C was screened by MTT method. (2) The optimal concentration of vitamin C and VK3 The drug concentration is 10 μM. (3) Flow cytometry to detect the apoptosis rate of each group The results showed that: 10μMVK3 + 1000μMVC combination group and 1000μMVC group, can significantly increase the rate of apoptosis (p <0.05). (4) Flow cytometry showed that the effect of VC + K3 on apoptosis was concentrated in S phase (p <0.05). Conclusion The combination of vitamin K3 and vitamin C can enhance the apoptosis of human bladder cancer T24 cells. The mechanism may be related to the block of T24 cell cycle of vitamin C in S phase.