敲除Linc00152对丝裂霉素耐药胃癌细胞NCI-N87/MMC的化疗耐药性影响及机制  

Effect and mechanisms of LINC00152 knockdown on chemotherapy resistance in mitomycin-resistant gastric cancer NCI-N87/MMC cells

作　　者：吴明东[1] 叶洁桐[1] 朱蓓蕾[1] 叶芳敏[1] 汪望月 Ming-Dong Wu;Jie-Tong Ye;Bei-Lei Zhu;Fang-Min Ye;Wang-Yue Wang(Department of Pharmacy,Lishui City People’s Hospital,Lishui 323000,Zhejiang Province,China;Department of Gastroenterology,Zhejiang Provincial People’s Hospital,Hangzhou 310000,Zhejiang Province,China)

机构地区：[1]丽水市人民医院药学部,浙江省丽水市323000 [2]浙江省人民医院消化内科,浙江省杭州市310000

出　　处：《世界华人消化杂志》2021年第7期332-339,共8页World Chinese Journal of Digestology

基　　金：丽水市公益性技术应用研究计划项目,No.2019SJZC47.

摘　　要：背景长基因间非编码RNA 152(long intergenic noncoding RNA 152,Linc00152)在胃癌组织中高表达,且其能促进胃癌细胞增殖、迁移与侵袭,而Linc00152对胃癌化疗耐药的影响和机制并不清楚.目的探究Linc00152对人胃癌细胞系NCI-N87化疗耐药性的影响及相关的作用机制.方法实时定量荧光聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,Real-time PCR)检测人胃癌细胞系NCI-N87及其丝裂霉素(mitomycin,MMC)耐药细胞系NCI-N87/MMC中Linc00152的表达情况.采用小分子RNA干扰技术敲除NCI-N87/MMC中Linc00152的表达后,MTT法检测细胞对MMC和顺铂的敏感性,流式细胞术检测细胞凋亡,Western Blot检测B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、半胱氨酸天冬氨酸蛋白酶-3(cysteine aspartic acid proteinase 3,caspase3)和劈裂的caspase3(cleaved-caspase3)的蛋白表达水平.此外,Real-time PCR和Western blot检测多药耐药蛋白1/P-糖蛋白(multidrug resistant protein 1/P-glycoprotein,MDR1/P-gp)、层粘连蛋白受体1前体抗原(P37-kDa laminin receptor-1 precursor antigen,Mgr1-Ag)以及多药耐药相关蛋白(multidrug resistance-associated protein,MRP)的表达.结果NCI-N87/MMC细胞中Linc00152的表达水平明显高于其亲本细胞NCI-N87.在NCI-N87/MMC细胞中,敲除Linc00152可诱导细胞凋亡,并增加其对MMC和顺铂的敏感性.敲除Linc00152可抑制NCI-N87/MMC细胞中Bcl-2蛋白表达,促进Bax蛋白表达和caspase 3的活化.此外,敲除Linc00152还可下调NCI-N87/MMC细胞中多药耐药基因MDR1、Mgr1-Ag、MRP及其编码的蛋白的表达.结论下调NCI-N87/MMC细胞中Linc00152的表达可提高细胞对MMC和顺铂的敏感性,其机制可能与调控细胞凋亡相关因子促进凋亡,同时下调细胞中多药耐药基因MDR1、Mgr1-Ag及MRP的表达有关.BACKGROUND Long intergenic noncoding RNA 152(LINC00152)is highly expressed in gastric cancer tissues,and it can promote the proliferation,migration,and invasion of gastric cancer cells.However,the effects and mechanisms of LINC00152 on chemotherapy resistance in gastric cancer are not clear.AIM To explore the effects and related mechanisms of LINC00152 on chemotherapy resistance in human gastric cancer cell line NCI-N87.METHODS The expression of LINC00152 in human gastric cancer cell line NCI-N87 and mitomycin(MMC)resistant cell line NCI-N87/MMC was detected by real-time PCR.After the expression of LINC00152 in NCI-N87/MMC cells was knocked down by RNA interference method,the sensitivity of cells to MMC and cisplatin was measured by MTT assay,cell apoptosis was detected by flow cytometry,and the protein expression levels of Bcl-2,Bax,Caspase 3,and cleaved Caspase 3 were determined by Western Blot.Furthermore,the expression levels of MDR1/P-gp,Mgr1-Ag,and MRP were evaluated by real-time PCR and Western Blot.RESULTS The expression level of INC00152 in NCI-N87/MMC cells was significantly higher than that in maternal NCI-N87 cells.LINC00152 knockdown induced apoptosis and increased sensitivity to MMC and cisplatin in NCI-N87/MMC cells.LINC00152 knockdown inhibited the expression of Bcl-2 protein in NCI-N87/MMC cells,but promoted the expression of Bax protein and the activation of Caspase 3.Furthermore,LINC00152 knockdown down-regulated the mRNA and protein expression of MDR1,Mgr1-Ag,and MRP in NCI-N87/MMC cells.CONCLUSION Down-regulation of LINC00152 in NCI-N87/MMC cells can increase the sensitivity of cells to MMC and cisplatin,and the mechanisms may be related to the promotion of cell apoptosis by regulating apoptotic-related factors,and down-regulation of MDR1,Mgr1-Ag,and MRP.

关 键 词：Linc00152 胃癌 NCI-N87细胞 丝裂霉素耐药 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]