KDM2A对人卵巢癌顺铂耐药细胞株A2780增殖和凋亡的影响及机制研究

Effects of KDM2A on the Resistance of Human Ovarian Cancer A2780 Cells to Cisplatin and Its Mechanisms

目的:探讨赖氨酸脱甲基酶2A(Lysine-specific demethylase 2A,KDM2A)对顺铂(cisplatin,DDP)耐药的人卵巢癌细胞A2780细胞增殖和凋亡的影响及其可能作用机制。方法:通过构建慢病毒载体转染A2780/DDP,分为A2780、A2780/DDP、A2780/DDP/KDM2A(转染KDM2A)、A2780/DDP/Jagged1(转染Jagged1)以及A2780/DDP/NC(转染病毒载体)组。采用Western blot检测3组KDM2A、Jagged1、Bcl2和BAX蛋白表达,CCK8和平板克隆形成实验检测细胞对顺铂的敏感性,流式细胞术检测细胞凋亡情况。结果:A2780/DDP细胞KDM2A和Jagged1的蛋白表达水平均显著高于A2780细胞(P<0.05),且A2780/DDP/KDM2A细胞中KDM2A和Jagged1的蛋白表达均低于A2780/DDP以及阴性对照组A2780/DDP/NC(P<0.05);A2780/DDP/Jagged1细胞的Jagged1蛋白表达低于A2780/DDP以及A2780/DDP/NC(P<0.05),而其KDM2A蛋白的表达比较差异无统计学意义(P>0.05)。不同浓度DDP处理的A2780/DDP/KDM2A细胞的生长抑制率均显著高于A2780/DDP/NC和A2780/DDP细胞(P<0.05),A2780/DDP/KDM2A细胞克隆形成数量亦明显高于A2780/DDP/NC和A2780/DDP细胞(P<0.05)。A2780/DDP/KDM2A细胞凋亡率为(25.84±3.27)%,明显高于A2780/DDP细胞[(14.29±1.96)%](P<0.05)和A2780/DDP/NC细胞[(12.46±2.15)%](P<0.05)。A2780/DDP/KDM2A细胞中Bcl-2蛋白表达明显低于A2780/DDP细胞(P<0.05),而A2780/DDP/KDM2A细胞Bax的表达水平却高于A2780/DDP细胞(P<0.05)。结论:KDM2A可能通过上调Jagged1的表达,促进人卵巢癌细胞A2780的增殖并抑制其凋亡,进而降低人卵巢癌耐药细胞A2780的顺铂敏感性。

Objective： To investigate the effect of lysine emethylase 2A on the proliferation and apoptosis of human ovarian cancer A2780 cells with cisplatin resistance and its potential mechanism. Methods： A2780/DDP was transfected by lentivirus vector including A2780/DDP/KDM2A,A2780/DDP/Jagged1 and A2780/DDP/NC cells. The expression of KDM2A,Jagged1,Bcl2 and BAX protein in 3 groups was detected by Western blot. The sensitivity of cells to cisplatin was compared by CCK8 and plate clone assay. The flow cytometry was used to detect the apoptosis in 3 groups. Results： The expression levels of KDM2A and Jagged1 were significantly higher in A2780/DDP cells than those in A2780 cells. The expression levels of KDM2A and Jagged1 proteins were significantly lower in A2780/DDP/KDM2A cells than those in A2780/DDP and A2780/DDP/NC cells（P〈0.05）. The expression of Jagged1 was significantly lower in A2780/DDP/Jagged1 cells than those in A2780/DDP cells（P〈0.05）,and the expression of KDM2A in A2780/DDP/Jagged1 cells showed no significant differences in comparison with that in A2780/DDP and A2780/DDP/NC cells（P〉 0.05）. The inhibition rate of A2780/DDP/KDM2A cells treated with different concentrations of DDP was significantly higher than that of A2780/DDP and A2780/DDP/NC cells（P〈0.05）. Moreover,the cell formation clones of A2780/DDP/KDM2A cells were also higher than those of A2780/DDP and A2780/DDP/NC cells（P〈0.05）. The 10 μmol/L DDP induced apoptotic rate of A2780/DDP/KDM2A cells [（25.84±3.27）%] was significantly higher than that of A2780/DDP [（14.29±1.96）%]（P〈0.05） and A2780/DDP/NC cells [（12.46±2.15）%]（P〈0.05）. The expression of Bcl2 in A2780/DDP/KDM2A cells was significantly lower than that of A2780/DDP cells（P〈0.05）,but the expression of Bax in A2780/DDP/KDM2A cells was higher than that of A2780/DDP cells（P〈0.05）. Conclusion： KDM2A could upregulate the expression of Jagged1,facilitate the proliferation,inhibitedapoptosis and enhance the cisplatin sensitivity of human ovarian cancer cell line A2780.