微小RNA-101对胰腺癌PANC1细胞增殖、凋亡和侵袭的影响及其作用机制  

Effect and mechanism of microRNA-101 on proliferation, apoptosis and invasion of pancreatic cancer PANC1 cells

作　　者：王宏 马磊 Wang Hong;Ma Lei(Department of General Surgery,Shanxi Province Fenyang Hospital,Fenyang 032200,China;Department of Pathology,Shanxi Province Fenyang Hospital,Fenyang 032200,China)

机构地区：[1]山西省汾阳医院普外科,汾阳032200 [2]山西省汾阳医院病理科,汾阳032200

出　　处：《中华胰腺病杂志》2023年第5期360-365,共6页Chinese Journal of Pancreatology

摘　　要：目的探讨微小RNA-101(miR-101)对胰腺癌PANC1细胞增殖、凋亡和侵袭的影响及其可能的作用机制。方法取对数生长期PANC1细胞分为5组,除对照组外,分别为转染miRNA模拟体的阴性对照(mimic-NC)组、转染miRNA-101模拟体(miR-101 mimic)组、转染miRNA抑制剂的阴性对照(inhibitor-NC)组、转染miRNA-101抑制剂(miR-101 inhibitor)组。采用qRT-PCR检测miR-101相对表达水平,MTT法检测细胞增殖率,流式细胞仪检测细胞凋亡率,Transwell实验检测细胞侵袭能力,qRT-PCR和蛋白质免疫印迹法检测IL-6、JAK2、p-JAK2、STAT3、p-STAT3 mRNA蛋白的相对表达水平,双荧光素酶报告基因法检测miR-101与IL-6的靶向关系。结果转染48 h后,对照组、mimic-NC组、miR-101 mimic组、inhibitor-NC组、miR-101 inhibitor组PANC1细胞miR101表达水平分别为1.98±0.12、2.01±0.18、6.73±0.23、2.16±0.22、1.34±0.13;增殖率分别为(32.75±2.43)%、(33.17±2.77)%、(15.68±1.17)%、(31.57±2.65)%和(45.75±3.16)%;凋亡率分别为(3.82±0.57)%、(3.54±0.55)%、(28.61±0.78)%、(3.57±0.63)%、(1.03±0.62)%;穿膜细胞数分别为(125.82±3.55)、(132.17±4.28)、(58.83±3.24)、(128.77±5.06)、(248.42±5.64)个/高倍视野。miR-101 mimic组miR101表达水平和凋亡率显著高于对照组和mimic-NC组,增殖率和穿膜细胞数显著低于对照组和mimic-NC组,而miR-101 inhibitor组miR-101表达水平和凋亡率显著低于对照组、inhibitor-NC组、miR-101 mimic组,增殖率和穿膜细胞数显著高于对照组、inhibitor-NC组、miR-101 mimic组。miR-101 mimic组PANC1细胞IL-6、JAK2、STAT3 mRNA和蛋白质表达水平显著低于对照组、mimic-NC组,而miR-101 inhibitor组显著高于对照组、miR-101 mimic组、inhibitor-NC组。以上差异均有统计学意义(P值均<0.001)。双荧光素酶报告基因法检测结果显示,miR-101可靶向作用IL-6。结论miR-101能够抑制胰腺癌细胞的增殖和侵袭,促进其凋亡,其机制可能是通过调节IL-6/JAK2/STAT3信号通路Objective To investigate the effect of microRNA-101(miR-101)on proliferation,apoptosis and invasion of PANC1 cells of pancreatic cancer and its potential mechanism.Methods PANC1 cells in logarithmic growth period were divided into 5 groups,including transfected miRNA mimic negative control group(mimic-NC group),transfected miRNA-101 mimic group(miR-101 mimic group),transfected miRNA inhibitor negative control group(inhibitor-NC group),and transfected miR-101 inhibitor group(miR-101 inhibitor group)besides control group.The relative expression level of miR-101 was detected by qRT-PCR,cell proliferation rate was determined by MTT assay,cell apoptosis rate was detected by flow cytometry,and cell invasion ability was detected by Transwell assay.The relative expression levels of IL-6,JAK2,p-JAK2,STAT3 and p-STAT3 mRNA and proteins were detected by qRT-PCR and Western blot.The targeting relationship between miR-101 and IL-6 was detected by dual luciferase reporter gene method.Results After 48 h of transfection,the expression levels of miR-101 in PANC1 cells in control group,mimic-NC group,miR-101 mimic group,inhibitor-NC group and miR-101 inhibitor group were 1.98±0.12,2.01±0.18,6.73±0.23,2.16±0.22 and 1.34±0.13;the proliferation rates were(32.75±2.43)%,(33.17±2.77)%,(15.68±1.17)%,(31.57±2.65)%and(45.75±3.16)%,respectively;the apoptosis rates were(3.82±0.57)%,(3.54±0.55)%,(28.61±0.78)%,(3.57±0.63)%and(1.03±0.62)%,respectively;the number of penetrating cells was(125.82±3.55),(132.17±4.28),(58.83±3.24),(128.77±5.06),(248.42±5.64)/high power field,respectively.The expression level of miR-101 and apoptosis rate in miR-101 mimic group were significantly higher than those in control group and mimic-NC group,and the proliferation rate and the number of transmembrane cells were significantly lower than those in control group and mimic-NC group.The expression level of miR-101 and apoptosis rate in miR-101 inhibitor group were significantly lower than those in control group,inhibitor-NC group and miR-101 mimi

关 键 词：胰腺癌 微小RNA-101 侵袭 凋亡 白介素-6 

分 类 号：R735.9[医药卫生—肿瘤]