miRNA-34a反义寡核苷酸对非小细胞肺癌细胞HCC827的影响  

The effect of miRNA-34a antisense oligonucleotide on non-small cell lung cancer cell line HCC827

作　　者：任丽坤 王瑜 庞世杰 范立华 Ren Likun;Wang Yu;Pang Shijie;Fan Lihua(Department of Comprehensive Oncology Therapy,Handan Second Hospital,Handan 056001,China;Department of Oncology,Handan Iron.and steel hospital,Handan 056002,China)

机构地区：[1]邯郸市第二医院肿瘤综合治疗科,056001 [2]邯钢医院肿瘤科,邯郸056002

出　　处：《中国医师杂志》2020年第2期211-215,共5页Journal of Chinese Physician

基　　金：河北省科技计划项目(182777144)。

摘　　要：目的探讨miRNA-34a反义寡核苷酸对非小细胞肺癌细胞的影响及分子机制。方法qRT-PCR检测人非小细胞肺癌细胞株HCC827和人正常肺细胞MRC-5中miRNA-34a表达水平。将HCC827细胞分为3组:空白对照组、阴性对照miRNA组(miRNA-NC)、反义寡核苷酸miRNA-34a组(脂质体2000转染反义寡核苷酸miRNA-34a);CCK-8法检测细胞增殖能力,吉姆萨染色检测细胞克隆能力,Transwell实验检测细胞迁移和侵袭能力;RT-PCR和Western blot检测细胞中磷酸酶和紧张素同系物(PTEN)、磷酸化蛋白激酶B(p-Akt)、磷脂酰肌醇3-激酶(PI3K) mRNA和蛋白表达水平。结果 HCC827细胞miRNA-34a相对表达量明显高于人正常肺细胞,差异有统计学意义(P <0. 01)。反义寡核苷酸miRNA-34a组miRNA-34a相对表达量明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P <0. 05);阴性对照miRNA组与空白对照组miRNA-34a相对表达量之间差异无统计学意义(P> 0. 05)。培养48、72、96 h时反义寡核苷酸miRNA-34a组HCC827细胞增殖水平明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P <0. 05)。反义寡核苷酸miRNA-34a组细胞克隆形成率明显低于阴性对照miRNA组、空白对照组,差异有统计学意义(P <0. 01)。反义寡核苷酸miRNA-34a组HCC827细胞迁移和侵袭个数明显低于阴性对照miRNA组和空白对照组,差异有统计学意义(P <0. 01)。反义寡核苷酸miRNA-34a组细胞PTEN mRNA和蛋白相对表达量明显高于阴性对照miRNA组与空白对照组,p-Akt、PI3K mRNA和蛋白相对表达量明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P <0. 05)。结论人非小细胞肺癌细胞中miRNA-34a表达水平明显高于人正常肺细胞;miRNA-34a反义寡核苷酸能够抑制人非小细胞肺癌细胞的增殖、克隆、迁移和侵袭,其机制可能与负调控PTEN/p-Akt/PI3K信号通路有关。Objective To investigate the effect of antisense oligonucleotides of miRNA-34a on non-small cell lung cancer(NSCLC)and its molecular mechanism.Methods The expression of miRNA-34a in human non-small cell lung cancer cell line HCC827 and human normal lung cell MRC-5 was detected by real time fluorescence quantitative polymerase chain reaction(qRT-PCR).HCC827 cells were divided into three groups:blank control group,negative control group,anti-sense oligonucleotide group(liposome 2000 transfected anti-sense oligonucleotide miRNA-34a);cell counting kit-8(CCK-8)method was used to detect cell proliferation,Jimsa staining was used to detect cell cloning ability,Transwell test was used to detect cell migration and invasion ability;RT-PCR and Western blot were used to detect phosphatase and ten-sin homolog(PTEN),phosphorylation-protein kinase B(p-Akt),phosphatidylinositol-3-kinase(PI3K)mRNA and protein expression.Results The relative expression of miRNA34a in HCC827 cells was significantly higher than that in human normal lung cells(P<0.01).The relative expression of miRNA34a in antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group(P<0.05),and there was no significant difference between negative control group and blank control group(P>0.05).At 48 h,72 h and 96 h,the proliferation level of HCC827 cells in anti-sense oligonucleotide miRNA-34a group was significantly lower than that in negative control group and blank control group(P<0.05).The cell cloning rate of antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group(P<0.01).The number of migration and invasion of HCC827 cells in antisense oligonucleotide RNA-34a group was significantly lower than that in negative control group and blank control group(P<0.01).The relative expression of PTEN mRNA and protein in antisense oligonucleotide miRNA-34a group was significantly higher than that in negative control group and blank control group

关 键 词：寡核苷酸类 反义 miRNA-34a 癌 非小细胞肺 细胞系 肿瘤 

分 类 号：R734.2[医药卫生—肿瘤]