长链非编码RNA缺氧诱导因子-1α-反义链2在肝癌中的表达及对肝癌增殖和侵袭能力的影响  

Expression of long non-coding RNA hypoxia-inducible factor-1αantisense RNA-2 in hepatocellular carcinoma and its effect on proliferation and invasion

作　　者：付雪岩 吴刚[1] 王雅玮[2] 王文青 Fu Xueyan;Wu Gang;Wang Yawei;Wang Wenqing(Department of General Surgery,the First Affiliated Hospital of China Medical University,Shenyang 110001,China;Department of Geriatric Surgery,the First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院肝胆外科暨器官移植科,沈阳110001 [2]中国医科大学附属第一医院老年外科,沈阳110001

出　　处：《中华实验外科杂志》2019年第5期837-840,共4页Chinese Journal of Experimental Surgery

基　　金：沈阳市科技局人口与卫生研究项目(F15-139-9-25);辽宁省自然科学基金(201520529).

摘　　要：目的检测长链非编码RNA缺氧诱导因子-1α-反义链2(lncRNA-HIF1A-AS2)在肝癌组织及肝癌细胞株中的表达并探讨其对肝癌增殖和侵袭能力的影响。方法采用实时荧光定量聚合酶链反应(FQ-PCR)技术检测HIF1A-AS2在49例肝癌患者的癌组织及其癌旁组织以及4种肝癌细胞株(Huh7、HepG2、SMMC-7721、Bel-7402)和正常肝细胞L02的表达,并分析HIF1A-AS2表达与临床病理特征之间的联系;选取两种高表达HIF1A-AS2肝癌细胞分别转染小干扰RNA(si-HIF1A-AS2)和阴性对照(si-NC);利用细胞增殖-毒性检测实验(CCK-8)和平板克隆实验检测细胞增殖能力;利用Transwell实验检测细胞侵袭能力。结果HIF1A-AS2在肝癌组织中的表达高于癌旁组织(-12.060±0.657比-12.450±0.683,P<0.01);HIF1A-AS2表达与年龄、肿瘤大小和肿瘤分化相关(χ2=4.573、4.871、8.353,P<0.05);在肝癌细胞株的表达也明显高于正常肝细胞株,其中在Huh7和HepG2的表达最高(Huh7:5.349±0.992,HepG2:5.660±0.726,P<0.01);CCK-8和平板克隆显示转染si-HIF1A-AS2后,细胞增殖能力明显低于对照组(Huh7:0.668±0.067比0.558±0.055,0.931±0.881比0.670±0.072,1.769±0.079比1.036±0.074;HepG2:1.063±0.060比0.803±0.097,1.767±0.091比1.083±0.080,P<0.01)(Huh7:542.70±25.50比378.00±19.31,HepG2:530.00±14.53比217.30±18.15,P<0.01);侵袭实验显示转染si-HIF1A-AS2后,细胞侵袭数明显低于对照组[Huh7:(116.300±6.506)个比(52.000±9.539)个,HepG2:(88.330±5.686)个比(55.000±5.245)个,P<0.01]。结论HIF1A-AS2在肝癌及细胞株中高表达,下调HIF1A-AS2表达可以抑制肝癌细胞的增殖和侵袭能力。Objective To detect the expression of long non-coding RNA hypoxia-inducible factor-1αantisense RNA-2(lncRNA-HIF1A-AS2)in hepatocellular carcinoma(HCC)and cell lines and its effect on proliferation and invasion of hepatoma cells.Methods Real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)was used to detect the expression of HIF1A-AS2 in 49 HCC tissues and adjacent normal tissues,four HCC cell lines(Huh7,HepG2,SMMC-7721,Bel-7402)and one normal liver cell line L02,then analyze the correlation between HIF1A-AS2 expression and clinicopathological features;Two high-expression HIF1A-AS2 HCC cells were transfected into small interfering RNA(si-HIF1A-AS2)and negative control(si-NC);Cell counting kit 8(CCK-8)assay and Colony-forming assay were conducted to evaluate the cell proliferation;Transwell assay was conducted to evaluate the cell invasion.Results The expression of HIF1A-AS2 in HCC tissues was higher than that in adjacent tissues(-12.060±0.657 vs.-12.450±0.683,P<0.01);The expression of HIF1A-AS2 was correlated with age,tumor size and tumor differentiation(χ2=4.573,4.871,8.353,P<0.05);The expression of HIF1A-AS2 in HCC cell line was also significantly higher than that in normal liver cell line,and the expression was highest in Huh7 and HepG2(Huh7:5.349±0.992,HepG2:5.660±0.726,P<0.01);CCK8 assay and colony-forming assay showed that the cell proliferation ability after transfection of si-HIF1A-AS2 was significantly lower than that of the control group in Huh7 and HepG2(Huh7:0.668±0.067 vs.0.558±0.055,0.931±0.881 vs.0.670±0.072,1.769±0.079 vs.1.036±0.074;HepG2:1.063±0.0604 vs.0.803±0.097,1.767±0.091 vs.1.083±0.080,P<0.01)(Huh7:542.70±25.50 vs.378.00±19.31,HepG2:530.00±14.53 vs.217.30±18.15,P<0.01);Transwell assay showed that the number of cell invasion was significantly lower after transfection of si-HIF1A-AS2[Huh7:(116.300±6.506)cells vs.(520.000±9.539)cells,HepG2:(88.330±5.686)cells vs.(55.000±5.245)cells,P<0.01].Conclusion HIF1A-AS2 is highly expressed in HCC and cell lines,and

关 键 词：肝细胞癌 长链非编码RNA-缺氧诱导因子-1α-反义链2 增殖 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]