川芎嗪基于KDM2B调控卵巢癌细胞顺铂耐药的机制
Mechanism of ligustrazine regulating cisplatin resistance in ovarian cancer cells based on KDM2B
蔡月红 1麦燕 1钱沁佳1
作者信息
- 1. 三亚中心医院妇产科,三亚 572000
- 折叠
摘要
目的 探索川芎嗪调控卵巢癌细胞顺铂耐药的潜在机制.方法 用顺铂(cisplatin,DDP)诱导A2780细胞成为A2780顺铂耐药细胞株(A2780/DDP);将其分为对照组、阴性对照组、干扰组和川芎嗪组.干扰组和阴性对照组分别用si-KDM2B和si-NC进行转染.川芎嗪组给予终浓度为5 nmol∙L-1的川芎嗪(培养基溶解)处理细胞,对照组给予相同体积的培养基.每组细胞培养48 h后,收集耐药细胞.实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,qRT-PCR)用于检测mRNA表达水平,噻唑蓝(methyl thiazolyl tetrazolium,MTT)用于细胞增殖实验,流式细胞术用于检测细胞凋亡,Western blotting用于检测细胞凋亡相关蛋白[(B淋巴细胞瘤-2(B-cell lymphoma-2,BCl-2)、B淋巴细胞瘤-2相关蛋白X(BCL2-associated X protein,Bax)和半胱氨酸蛋白酶-3(caspase-3)]的表达水平.结果 赖氨酸特异性去甲基化酶2B(lysine-specific demethylase 2B,KDM2B)在A2780/DDP中高表达,A2780/DDP细胞经川芎嗪或者KDM2B敲减处理后可以抑制A2780/DDP细胞增殖、促进细胞凋亡、上调凋亡相关蛋白(Bax和caspase-3)和下调BCl-2的表达水平.结论 川芎嗪可能通过调控KDM2B抑制A2780/DDP细胞增殖和促进细胞凋亡.
Abstract
Objective To explore the potential mechanism of ligustrazine regulating cisplatin resistance in ovarian cancer cells.Methods Cisplatin(DDP)was used to induce A2780 to become A2780 cisplatin-resistant cell lines(A2780/DDP).This study was divided into control group,negative control group,interference group,and ligustrazine group.The interference group and the negative control group were transfected with si-KDM2B and si-NC,respectively.The ligustrazine group was given a final concentration of 5 nmol∙L-1 ligustrazine(dissolved in the medium)to treat the cells,and the control group was given the same volume of the medium.After each group of cells were cultured for 48 hours,the drug-resistant cells were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect mRNA expression levels,methyl thiazolyl tetrazolium(MTT)was used for exploring cell proliferation,flow cytometry was carried out for detecting apoptosis,and Wes-tern blotting was used for detecting the expression levels of apoptosis-related proteins[B-cell lymphoma-2(BCl-2),BCL2-as-sociated X protein(Bax)and caspase-3].Results Lysine-specific demethylase 2B(KDM2B)was highly expressed in A2780/DDP.After treated with ligustrazine or knockdown of KDM2B,A2780/DDP cells inhibited cell proliferation,promoted cell apoptosis,up-regulated apoptosis-related proteins(Bax and caspase-3)and down-regulated BCl-2 expression level in A2780/DDP.Conclusion Ligustrazine may inhibit A2780/DDP cell proliferation and promote cell apoptosis by KDM2B.
关键词
川芎嗪/赖氨酸特异性去甲基化酶2B/卵巢癌/A2780顺铂耐药细胞株Key words
ligustrazine/KDM2B/ovarian cancer/A2780/DDP引用本文复制引用
基金项目
海南省卫生计生行业科研计划(18A200164)
出版年
2024
NETL
NSTL
维普
万方数据