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

作　　者：蔡月红 麦燕 钱沁佳 CAI Yuehong;MAI Yan;QIAN Qinjia(Department of Obstetrics and Gynecology,Sanya Central Hospital,Sanya 572000,China)

机构地区：[1]三亚中心医院妇产科,三亚572000

出　　处：《西北药学杂志》2024年第1期61-65,共5页Northwest Pharmaceutical Journal

基　　金：海南省卫生计生行业科研计划项目(编号:18A200164)。

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 demethylase2B,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。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 Western blotting was used for detecting the expression levels of apoptosis-related proteins[B-cell lymphoma-2(BCl-2),BCL2-associated 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 

分 类 号：R965[医药卫生—药理学]