黏蛋白1基因小干扰RNA联合丙泊酚抑制肺癌H460细胞生长的机制研究  

Study on the mechanism of MUC1 gene siRNA combined with propofol inhibiting the growth of lung cancer H460 cells

作　　者：陆巍 王飞[2] 梁柳芬 LU Wei;WANG Fei;LIANG Liufen(Outpatient Department of Beijiao Hospital,Shunde District,Foshan City,Foshan,Guangdong 528300,China;Department of Thoracic Surgery,First People's Hospital of Foshan City,Foshan,Guangdong 528300,China)

机构地区：[1]佛山市顺德区北滘医院门诊部,广东佛山528300 [2]佛山市第一人民医院胸外科,广东佛山528300

出　　处：《安徽医药》2021年第3期441-445,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨黏蛋白1(MUC1)基因小干扰RNA(siRNA)联合丙泊酚对肺癌H460细胞生长的抑制及可能的机制。方法siRNA干扰人肺癌H460细胞株MUC1基因,采用实时荧光定量PCR(qPCR)和蛋白质印迹法(Western blotting)法检测转染效果;以不同浓度的丙泊酚干预H460细胞,细胞计数试剂盒(CCK-8)检测丙泊酚对H460细胞活力的影响,筛选丙泊酚作用浓度;实验分为转染对照(si-NC)组、转染MUC1 siRNA(si-MUC1)组、丙泊酚干预(Propofol)组和转染MUC1 siRNA联合丙泊酚干预(si-MUC1+Propofol)组;采用CCK-8检测H460细胞增殖情况,克隆形成实验分析H460细胞克隆形成能力,Western blotting检测H460细胞中增殖细胞核抗原(PCNA)和细胞周期蛋白D1(cyclin D1)、蛋白激酶B(protein kinase B,Akt)和磷酸化Ak(t p-Akt)的表达水平。结果si-NC组和si-MUC1组细胞中MUC1 mRNA的表达分别为:(1.00±0.10)、(0.18±0.02),MUC1蛋白表达分别为:(0.56±0.06)、(0.21±0.02),转染MUC1 siRNA的H460细胞中MUC1 mRNA和蛋白表达水平显著降低(t=24.122,P<0.001,t=16.602,P<0.001);不同浓度的丙泊酚对H460细胞有不同程度的抑制作用,并筛选出半数致死浓度55.96μmol/L的丙泊酚用于后续实验;si-NC组、si-MUC1组、Propofol组和si-MUC1+Propofol组细胞克隆形成数分别为:(163.36±8.22)个、(101.26±7.69)个、(92.64±6.88)个、(52.09±6.14)个,si-MUC1组和Propofol组细胞活力、克隆形成能力、PCNA、cyclin D1和p-Akt的表达水平均显著低于si-NC组(P<0.05),高于si-MUC1+Propofol组(P<0.05)。结论敲低MUC1基因和丙泊酚均能抑制肺癌H460细胞生长,两者联合对H460细胞生长抑制作用更显著,其作用机制可能与抑制Akt信号通路的激活有关。Objective To investigate the inhibitory effect of mucin 1(MUC1)gene small interfering RNA(siRNA)combined with propofol on the growth of lung cancer H460 cells and its possible mechanism.Methods siRNA interfered with the MUC1 gene of hu⁃man lung cancer H460 cell line.Real-time fluorescent quantitative PCR(qPCR)and Western blotting(Western blotting)were used to detect the transfection effect;different concentrations of propofol were used to intervene in H460 cells.Cell counting kit(CCK-8)De⁃tect the effect of propofol on the viability of H460 cells,and screen the concentration of propofol;the experiment was divided into trans⁃fection control(si-NC)group,transfection MUC1 siRNA(si-MUC1)group,propofol intervention(Propofol))Group and transfected MUC1 siRNA combined with propofol intervention(si-MUC1+Propofol)group;CCK-8 was used to detect the proliferation of H460 cells,the clone formation experiment was used to analyze the cloning ability of H460 cells,and the proliferating cell nuclear antigen in H460 cells was detected by Western blotting(PCNA)and cyclin D1(cyclin D1),protein kinase B(protein kinase B,Akt)and phosphor⁃ylated Akt(p-Akt)expression levels.Results The expression of MUC1 mRNA in the cells of the si-NC group and si-MUC1 group were:(1.00±0.10),(0.18±0.02),and the MUC1 protein expression were:(0.56±0.06),(0.21±0.02),transfection The expression levels of MUC1 mRNA and protein in H460 cells of MUC1 siRNA were significantly reduced(t=24.122,P<0.001,t=16.602,P<0.001);different concentrations of propofol have different degrees of inhibition on H460 cells,and screening Propofol with a lethal concentration of 55.96μmol/L was used in subsequent experiments;the number of cell clones in the si-NC group,si-MUC1 group,Propofol group and si-MUC1+Propofol group were:(163.36±8.22)Cells,(101.26±7.69)cells,(92.64±6.88)cells,(52.09±6.14)cells,cell viability,clono⁃genic ability,PCNA,cyclin D1 and p-Akt expression levels in si-MUC1 and Propofol groups were significant Lower than si-NC group(P<0.05),higher

关 键 词：肺肿瘤 肿瘤细胞 培养的 MUC1基因 丙泊酚 肺癌细胞 生长 AKT信号通路 

分 类 号：R734.2[医药卫生—肿瘤]