吉非替尼通过靶向circ-0000745/miR-421轴调控胃癌细胞N87增殖、凋亡的分子机制  

The Molecular Mechanism of Gefitinib Regulating the Proliferation and Apoptosis of Gastric Cancer Cell N87 by Targeting the circ-0000745/miR-421 Axis

作　　者：胥陶 刘丽[2] 马玉蓉[1] 曹承刚 李召辉[1] Xu Tao;Liu Li;Ma Yurong;Cao Chenggang;Li Zhaohui(Department of Pharmacy,Chongqing Fifth People’s Hospital,Chongqing 400062,China;Department of Oncology,Chongqing Fifth People's Hospital;Department of Pharmacy,Rongchang District People's Hospital)

机构地区：[1]重庆市第五人民医院药剂科,重庆400062 [2]重庆市第五人民医院肿瘤科 [3]重庆市荣昌区人民医院药剂科

出　　处：《中国药师》2021年第12期2160-2166,共7页China Pharmacist

摘　　要：目的:探讨吉非替尼是否通过环状RNA(circRNA)circ-0000745/微小RNA-421(miR-421)轴调控胃癌细胞N87增殖、凋亡。方法:运用细胞计数试剂盒8(CCK-8)、平板克隆实验、蛋白印迹(Western blot)分析、流式细胞术与定量逆转录聚合酶链反应(qRT-PCR)测定空白对照(NC)组、0.125,0.25,0.5μmoL·L^(-1)吉非替尼组、si-NC组、si-circ-0000745组、NC+si-NC组、吉非替尼+si-NC组、吉非替尼+si-circ-0000745组、si-NC+anti-miR-NC组、si-circ-0000745+anti-miR-NC组、si-circ-0000745+anti-miR-421组胃癌细胞N87的活力、克隆形成、细胞周期蛋白D1(CyclinD1)、P21、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达、凋亡率与circ-0000745、miR-421表达。双荧光素酶实验分析circ-0000745与miR-421间的关系。结果:与NC组相比,0.125,0.25,0.5μmol·L^(-1)吉非替尼组细胞活力、克隆形成数、CyclinD1、Bcl-2蛋白表达量、miR-421表达量逐渐降低,而P21蛋白表达量、凋亡率、Bax蛋白表达量、circ-0000745表达量逐渐增加,且不同剂量组差异有统计学意义(P<0.05)。circ-0000745靶向负调控miR-421。与si-NC组相比,si-circ-0000745组胃癌细胞N87活力、克隆形成数、CyclinD1、Bcl-2蛋白表达量升高,P21蛋白表达量、凋亡率和Bax蛋白表达量降低,差异有统计学意义(P<0.05)。与吉非替尼+si-NC组相比,吉非替尼+si-circ-0000745组N87细胞中活力、克隆形成数、CyclinD1、Bcl-2蛋白表达量增加,P21、Bax蛋白表达量、凋亡率减少,差异均有统计学意义(P<0.05)。与si-circ-0000745+anti-miR-NC组相比,si-circ-0000745+anti-miR-421组N87细胞活力、克隆形成数、CyclinD1、Bcl-2蛋白表达量降低,P21、Bax蛋白表达量、凋亡率升高,差异有统计学意义(P<0.05)。结论:吉非替尼通过上调circ-0000745靶向miR-421,抑制胃癌细胞N87增殖,并诱导其凋亡。Objective: To investigate whether gefitinib regulates the proliferation and apoptosis of gastric cancer cell N87 through the circular RNA(circRNA) circ-0000745/microRNA-421(miR-421) axis. Methods: Using cell counting kit 8(CCK-8), plate cloning experiment, western blot analysis, flow cytometry and quantitative reverse transcription polymerase chain reaction(qRT-PCR) to determine the NC group, 0.125, 0.25, 0.5 μmol·L^(-1)Gefitinib group, si-NC group, si-circ-0000745 group, NC+si-NC group, Gefitinib+si-NC group, Gefitinib+si-circ-0000745 Group, si-NC+anti-miR-NC group, si-circ-0000745+anti-miR-NC group, si-circ-0000745+anti-miR-421 group gastric cancer cell N87 viability, clone formation, cyclin D1(CyclinD1), P21, B-cell lymphoma/leukemia-2(Bcl-2), Bcl-2 related X protein(Bax) expression, apoptosis rate and circ-0000745, miR-421 expression. The dual luciferase experiment analyzed the relationship between circ-0000745 and miR-421. Results: Compared with the NC group, the cell viability, the number of colonies, CyclinD1, Bcl-2 protein expression and miR-421 expression in 0.125, 0.25 and 0.5 μmol·L^(-1)gefitinib group gradually decreased, while P21 protein expression, apoptosis rate, Bax protein expression, and circ-0000745 expression gradually increased, and the difference between different dose groups was statistically significant(P<0.05). circ-0000745 targeted and negatively regulated the expression of miR-421. Compared with the si-NC group, the si-circ-0000745 group increased the viability of gastric cancer cell N87, the number of clones, the expression of CyclinD1 and Bcl-2 protein, and decreased the expression of P21 protein, the rate of apoptosis and the expression of Bax protein. The difference was statistically significant. Significance(P<0.05). Compared with the Gefitinib+si-NC group, the viability, the number of clones, the expression of CyclinD1 and Bcl-2 protein in the N87 cells of the Gefitinib+si-circ-0000745 group increased, and the expression of P21, Bax protein, and the apoptosis rate decreased.

关 键 词：吉非替尼 circ-0000745 miR-421 胃癌细胞N87 增殖 凋亡 

分 类 号：R965.1[医药卫生—药理学]