摘要
背景与目的:肝细胞肝癌是一种临床上常见的恶性肿瘤,由于其对化疗药物的耐受,常导致预后较差。microRNA(miRNA,miR)是一类长链非编码小RNA,参与肿瘤的多种生物学功能。miR-27a-3p作为代谢相关的miRNA被广泛地研究,而与化疗敏感性之间关系的报道较少。通过探讨miR-27a-3p参与肝癌化疗敏感性及其可能机制,为肝癌化疗提供新的理论依据。方法:应用癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析miR-27a-3p在肝细胞肝癌患者癌组织与癌旁组织中的表达。采用LipofectamineTM 3000脂质体瞬时转染方法,将miR-27a-3p过表达质粒和阴性对照质粒(miR-control)转染肝癌细胞株HepG2,将敲低质粒miR-inhibitor-27a-3p和阴性对照质粒(miR-inhibitor-control)转染PLC细胞,分别加入不同浓度的顺铂(cisplatin,DDP)处理48 h,采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测细胞增殖情况;采用流式细胞术检测细胞凋亡;蛋白[质]印迹法(Western blot)技术检测磷脂酰肌醇3激酶和蛋白激酶B(phosphoinosmde-3-kinase/protein kinase B,PI3K/AKT)信号通路相关的蛋白表达。结果:miR-27a-3p在肝癌组织中的表达明显低于癌旁组织(P<0.05);在HepG2过表达细胞中,MTT结果显示,实验组miR-27a+DDP细胞对DDP的半数抑制浓度(IC50)为(1.02±0.03)μg/mL,较对照组miR-control+DDP(1.27±0.08)μg/mL、空白对照组+DDP(1.73±0.08)μg/mL均降低(P<0.05);细胞凋亡分析结果显示,实验组miR-27a+DDP的凋亡率[(31.03±0.20)%]显著高于对照组+DDP[(18.51±2.96)%]和空白对照组+DDP[(8.73±1.58)%](P<0.05);Western blot检测结果显示,miR-27a+DDP组的PI3K表达水平(0.28±0.01)较DDP组[(0.43±0.01)]、miR-27a组(0.57±0.11)及miR-control组(0.72±0.01)降低(P<0.05);miR-27a+DDP组的p-AKT的表达水平(0.29±0.01)与DDP组(0.46±0.05)、miR-27a组(0.67±0.01)及miR-control组(0.10±0.01)相比明显降低(P<0.05);而miR-27a+DDP组的C-caspase的表达水平(0.69±0.01)则高于DDP组(0.51
Background and purpose:Hepatocellular carcinoma is a common malignant tumor in clinic.Because of its tolerance to chemotherapeutic drugs,it often leads to poor prognosis.MicroRNA(miRNA,miR)is a family of small long non-coding RNA molecules,and dysregulated miRNA is associated with various tumor biological functions.MicroRNA-27a-3p has been extensively studied as a metabolism-related miRNA,but there are few reports on the relationship between miRNA and chemosensitivity.This study was focused on the effect of miR-27a-3p on chemoresistance in hepatocellular carcinoma to provide novel theoretical basis for liver cancer chemotherapy.Methods:The Cancer Genome Atlas(TCGA)database was used to analyze the expression of miR-27a-3p in hepatocellular carcinoma and adjacent tissues.Using liposome transfection method,miR-27a-3p overexpression plasmid and negative control plasmid(miR-control)were transfected into hepatocellular carcinoma cell line HepG2.miR-inhibitor-27a-3p knockdown plasmid and negative control plasmid(miR-inhibitor-control)were transfected into PLC cells.Then the cells were treated with different concentrations of cisplatin(DDP)for 48 h.MTT assay was used to detect the proliferation,and flow cytometry was used to detect the apoptosis and cell cycle.Western blot technique was used to detect the protein expression in PI3k/AKT signaling pathway.Results:The expression of miR-27a-3p in hepatocellular carcinoma tissue was significantly lower than that of the adjacent tissues(P<0.05).In miR-27a overexpressed HepG2 cells,MTT results showed that the half inhibitory concentration(IC50)of DDP in miR-27a+DDP group[(1.02±0.03)μg/mL]was significantly decreased compared with miR-control+DDP group[(1.27±0.08)μg/mL]and the blank+DDP group[(1.73±0.08)μg/mL](P<0.05).The apoptosis results showed that miR-27a+DDP group[(31.03±0.20)%]had significantly increased apoptotic rate compared with miR-control+DDP group[(18.51±2.96)%]and the blank+DDP group[(8.73±1.58)%](P<0.05).Western blot results showed that the PI3K expression
作者
杨颖
杨志芳
才层
张瑞丽
路鹏霏
贾春丽
李志鹏
毛睿
张华
包永星
YANG Ying;YANG Zhifang;CAI Ceng;ZHANG Ruili;LU Pengfei;JIA Chunli;LI Zhipeng;MAO Rui;ZHANG Hua;BAO Yongxing(Department of Cancer Center,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang Uigur Autonomous Region,China)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2019年第10期788-794,共7页
China Oncology
基金
国家自然科学基金地区科学基金项目（81560388）
