KAI1基因对膀胱癌细胞株T24增殖和凋亡的影响及相关机制研究

The Research of KAI1 Gene in the Proliferation and Apoptosis of Bladder Cancer Cell Line T24 and the Related Mechanism

摘要: 目的：探讨KAI1对膀胱癌细胞株T-24的增殖和凋亡的影响及相关机制。方法：采用慢病毒感染的方法，使T24细胞株过表达KAI1基因，QPCR及细胞组织化学技术检测T24转染后KAI1的基因及蛋白表达量。MTT技术检测T24细胞的增殖能力；流式细胞技术检测细胞周期及T24细胞凋亡情况；Western blotting检测CyclinD1、caspase-3蛋白表达。结果：慢病毒感染后，T24细胞株的KAI1的mRNA转录水平及蛋白的表达水平显著增强(P < 0.01)；细胞增殖能力明显低于空载组及空白对照组(P < 0.01)；细胞凋亡能力明显高于空载组及空白对照组(P < 0.01)；细胞周期阻滞于G0/G1期，S期细胞明显减少(P < 0.01)；CyclinD1蛋白的表达明显下调，而caspase-3蛋白表达上调。结论：过表达KAI1可显著降低膀胱癌细胞株T24的增殖能力，增强细胞凋亡。而这种作用是通过降低细胞内CyclinD1蛋白表达以及上调caspase-3蛋白实现的。

Abstract: Objective: To investigate how the KAI1 gene effect the proliferation and apoptosis of bladder cancer cell line T24 and the related mechanism. Methods: T24 cell line were over-expressed KAI1 gene by lentivirus infection. The KAI1 expression in infected T24 cell were detected by QPCR and cell histochemical technique. MTT technique was used to detect the proliferation ability of T24 cells. The cell cycle and apoptosis of T24 cells were detected by flow cytometry. The expression of CyclinD1 and caspase-3 protein was detected by Western blotting. Results: After virus infection, the expression level of mRNA and protein in T24 cell line KAI1 was significantly increased (P < 0.01); cell proliferation was significantly lower than that of no load group and blank control group (P < 0.01); cell apoptosis was significantly higher than those in no load group and blank control group (P < 0.01); cell cycle arrest in G0/G1 phase, S phase cells significantly decreased (P < 0.01); the expression of CyclinD1 protein was down regulated, while caspase-3 protein expression was high regulated. Conclusion: overexpression of KAI1 can significantly reduce the proliferation of bladder cancer cell line T24, enhance cell apoptosis. This effect is achieved by reducing intracellular CyclinD1 protein expression and upregulation of caspase-3 protein.