慢病毒载体介导的长链非编码RNA Rpph1沉默对胆囊癌GBC-SD细胞增殖、细胞周期和凋亡的影响  

Effects of lentiviral vector-mediated lncRNA Rpph1 silencing on proliferation,cell cycle and apoptosis of gallbladder carcinoma GBC-SD cells

作　　者：杨隆良 郭虎林 马瑜 YANG Longliang;GUO Hulin;MA Yu(General Surgery,The Fifth People's Hospital of Qinghai,Xining,Qinghai 810000,China;Oncology,The Fifth People's Hospital of Qinghai,Xining,Qinghai 810000,China;Department of Hepatobiliary Surgery,Affiliated Hospital of Qinghai University,Xining,Qinghai 810000,China)

机构地区：[1]青海省第五人民医院普外科,青海西宁810000 [2]青海省第五人民医院肿瘤内科,青海西宁810000 [3]青海大学附属医院肝胆外科,青海西宁810000

出　　处：《安徽医药》2021年第9期1792-1796,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的研究长链非编码RNA(lncRNA)Rpph1对胆囊癌细胞增殖、细胞周期和凋亡的影响和潜在机制。方法以人胆囊上皮细胞HGBEC为对照,实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测胆囊癌细胞系GBC-SD、SGC-996和NOZ中lncRNARpph1的表达。将GBC-SD细胞分为对照组、si-con组、si-lncRpph1组。蛋白质印迹法检测周期蛋白依赖性激酶2(CDK2)、Ki-67、剪切型胱天蛋白酶3(Cleavedcaspase-3)、剪切型胱天蛋白酶9(Cleavedcaspase-9)、β连环素(β-catenin)和c-Myc蛋白表达水平,细胞计数试剂盒(CCK-8)法和流式细胞术分别检测细胞增殖、细胞周期和凋亡率。结果与对照相比,胆囊癌细胞系GBC-SD、SGC-996和NOZ组细胞中lncRNARpph1含量[(4.89±0.31)、(3.42±0.42)、(3.96±0.45)比(1.07±0.13)]显著升高(P<0.05);与si-con组比较,si-lncRpph1组GBC-SD细胞活力[(0.78±0.08)比(1.17±0.13)]降低,凋亡率[(12.89±1.96)%比(4.26±0.65)%]、G0~G1期细胞比例[(63.67±2.62)%比(52.57±1.32)%]升高,CDK2[(0.22±0.04)比(0.41±0.05)]、Ki-67[(0.31±0.04)比(0.62±0.04)]、β-catenin[(0.32±0.05)比(0.63±0.05)]和[c-Myc(0.21±0.04)比(0.41±0.06)]蛋白表达降低(P<0.05),Cleavedcaspase-3[(0.44±0.05)比(0.14±0.04)]和Cleavedcaspase-9[(0.54±0.05)比(0.17±0.03)]蛋白表达增加(P<0.05)。结论慢病毒载体沉默lncRNARpph1可能通过Wnt/β-catenin信号通路抑制胆囊癌GBC-SD细胞增殖,诱导细胞周期停滞和促进细胞凋亡。lncRNA Rpph1是胆囊癌的潜在分子靶点。Objective To investigate the effects of long non-coding RNA(lncRNA)Ribonuclease P RNA component H1(Rpph1)on the proliferation,cell cycle,and apoptosis of gallbladder carcinoma cells and the underlying mechanism.Methods Taking the human gallbladder epithelial cell HGBEC as the control,the expression levels of lncRNA Rpph1 in gallbladder carcinoma cell lines SGC-996,GBC-SD and NOZ were detected by quantitative real-time PCR(qRT-PCR).The GBC-SD cells were divided into control group,si-con group and si-lnc Rpph1 group.The expression levels of cyclin dependent kinase 2(CDK2),Ki-67,cleaved caspase-3,cleaved caspase-9,β-catenin and c-Myc proteins were detected by Western blotting.Cell proliferation,cell cycle and apoptosis rate were measured by cell counting kit(CCK-8)assay and flow cytometry,respectively.The relationship between lncRNA Rpph1 was validated by dual-luciferase reporter assay system.Results Compared with the control group,the levels of lncRNA Rpph1[(4.89±0.31),(3.42±0.42),(3.96±0.45)vs.(1.07±0.13)]in gallbladder carcinoma cell lines SGC-996,GBC-SD and NOZ were significantly increased,and the difference was statistically significant(P<0.05).Compared with the si-con group,the viability[(0.78±0.08)vs.(1.17±0.13)]of GBC-SD cell in the si-lnc Rpph1 group was reduced,and the cell apoptosis rate[(12.89±1.96)%vs.(4.26±0.65)%]and G0-G1 stage proportion[(63.67±2.62)%vs.(52.57±1.32)%]were increased,CDK2[(0.22±0.04)vs.(0.41±0.05)],Ki-67[(0.31±0.04)vs.(0.62±0.04)],β-catenin[(0.32±0.05)vs.(0.63±0.05)]and c-Myc[(0.21±0.04)vs.(0.41±0.06)]protein expression were decreased,cleaved caspase-3[(0.44±0.05)vs.(0.14±0.04)]and cleaved caspase-9[(0.54±0.05)vs.(0.17±0.03)]protein expression were increased,and the difference was statistically significant(P<0.05).Conclusions The silencing of lncRNA Rpph1 by lentiviral vector may inhibit the proliferation,induce cell cycle arrest and promote apoptosis of GBC-SD cells through Wnt/β-catenin signaling pathway.LncRNA Rpph1 is a potential molecular target for gallbl

关 键 词：胆囊肿瘤 长链非编码RNA Rpph1 慢病毒载体 增殖 细胞周期 凋亡 

分 类 号：R735.8[医药卫生—肿瘤]