慢病毒介导长链非编码RNA BACE2-IT1过表达抑制胃癌细胞的增殖和侵袭  

Lentivirus-mediated Long-chain Non-coding RNA BACE2-IT1 Overexpression Inhibits Proliferation and Invasion of Gastric Cancer Cells

作　　者：杨杰智 徐倩 杨明月 吕红琼 黄静 Yang Jiezhi;Xu Qian;Yang Mingyue(Department of Oncology,Luoyang Central Hospital Affiliated to Zhengzhou University,Henan 471000,China)

机构地区：[1]郑州大学附属洛阳中心医院肿瘤内科,洛阳471000 [2]郑州大学附属洛阳中心医院生殖助孕科,洛阳471000

出　　处：《医学研究杂志》2020年第1期163-167,共5页Journal of Medical Research

摘　　要：目的观察长链非编码RNA(long-chain non-coding RNA,lncRNA)BACE2-IT1在胃癌组织和细胞株中的表达,分析慢病毒介导BACE2-IT1过表达对胃癌细胞增殖和侵袭影响的作用机制。方法实时定量聚合酶链反应(qPCR)检测26例胃癌组织和5种胃癌细胞株中BACE2-IT1的表达。以BACE2-IT1表达最低的胃癌细胞株感染载有BACE2-IT1的慢病毒(实验组)和空载慢病毒(对照组)。通过CCK-8实验和Transwell侵袭实验分析慢病毒介导BACE2-IT1过表达对胃癌细胞株增殖能力和侵袭能力的影响。qPCR和蛋白质印迹法(Western blot法)检测过表达BACE2-IT1后HECT家族E3泛素连接酶1(HACE1)的表达水平及Wnt/β-catenin信号通路的活化程度。结果与癌旁组织比较,BACE2-IT1在胃癌组织中呈低表达(P<0.01)。与永生化胃黏膜上皮细胞比较,BACE2-IT1在5种胃癌细胞株中呈低表达(P<0.05),在BGC-823细胞中的表达量最低(P<0.01)。与对照组比较,实验组过表达BACE2-IT1在体外可抑制BGC-823细胞的增殖能力(P<0.05)。对照组和实验组侵袭细胞数分别为87.82±9.57和36.65±5.78,BACE2-IT1在体外可抑制BGC-823细胞的侵袭能力(P<0.01)。与对照组比较,BACE2-IT1过表达导致BGC-823细胞中HACE1在mRNA和蛋白水平的表达上调(P<0.01)。Wnt/β-catenin信号通路的活化被抑制。结论BACE2-IT1在胃癌组织和细胞株中均呈低表达,BACE2-IT1过表达能抑制胃癌BGC-823细胞的增殖能力和侵袭能力,其分子机制可能是促进HACE1蛋白的表达,抑制Wnt/β-catenin信号通路的活化。Objective To observe the expression of long-chain non-coding RNA(lncRNA)BACE2-IT1 in gastric cancer tissues and cell lines,and to analyze the mechanism of lentivirus-mediated BACE2-IT1 overexpression affecting proliferation and invasion of gastric cancer cells.Methods Real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of BACE2-IT1 in 26 cases of gastric cancer tissues and five gastric cancer cell lines.The gastric cancer cell line with the lowest BACE2-IT1 expression was infected with the lentivirus carrying BACE2-IT1(experimental group)and the empty lentivirus(control group).The effects of lentivirus-mediated BACE2-IT1 overexpression on the proliferation and invasion of gastric cancer cell lines were analyzed by CCK-8 assay and Transwell invasion assay.The expression level of HECT family E3 ubiquitin ligase 1(HACE1)and the activation level of Wnt/β-catenin signaling pathway were detected by qPCR and Western blot.Results BACE2-IT1 was down-regulated in gastric cancer tissues compared with adjacent tissues(P<0.01).Compared with immortalized gastric mucosal epithelial cells,BACE2-IT1 was down-regulated in five gastric cancer cell lines(P<0.05),and lowest in BGC-823 cells(P<0.01).Compared with the control group,overexpression of BACE2-IT1 in the experimental group inhibited the proliferation of BGC-823 cells in vitro(P<0.05).The number of invading cells in the control group and the experimental group were 87.82±9.57 and 36.65±5.78 respectively.BACE2-IT1 inhibited the invasive ability of BGC-823 cells in vitro(P<0.01).Overexpression of BACE2-IT1 resulted in up-regulation of HACE1 expression in mRNA and protein levels in BGC-823 cells compared to the control group(P<0.01).Activation of the Wnt/β-catenin signaling pathway was inhibited.Conclusion BACE2-IT1 is lowly expressed in gastric cancer tissues and cell lines.BACE2-IT1 overexpression can inhibit the proliferation and invasion of gastric cancer BGC-823 cells.The molecular mechanism may be to promote the expression of HACE1

关 键 词：胃癌 长链非编码RNA HECT家族E3泛素连接酶1 增殖 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]