lncRNA LAMA5-AS1通过上调TFPI2表达抑制鼻咽癌细胞的增殖和迁移

目的:检测鼻咽癌组织和细胞株中长链非编码RNA（long non-coding RNA，lncRNA）LAMA5-AS1的表达，观察lncRNA LAMA5-AS1对鼻咽癌细胞增殖和迁移的影响并探讨其作用机制。方法:荧光实时定量聚合酶链式反应（qPCR）分别检测lncRNA LAMA5-AS1在68例鼻咽癌组织及62例慢性鼻咽炎组织、鼻咽癌细胞株及永生化鼻咽上皮细胞中的相对表达。选择lncRNA LAMA5-AS1表达最少的细胞株，分别转染阴性对照质粒（对照组）或载有lncRNA LAMA5-AS1序列质粒（实验组）。MTS法和细胞划痕实验分别检测高表达lncRNA LAMA5-AS1对细胞增殖和迁移能力的影响。qPCR检测高表达lncRNA LAMA5-AS1对组织因子途径抑制物2（tissue factor pathway inhibitor 2，TFPI2）基因mRNA表达的影响，Western blotting检测TFPI2蛋白和ERK信号通路蛋白表达。结果:与慢性鼻咽炎组织比较，lncRNA LAMA5-AS1在鼻咽癌组织中表达明显降低（P＜0.01）。与人永生化鼻咽上皮细胞比较，lncRNA LAMA5-AS1在鼻咽癌细胞株中表达明显降低（P＜0.01），在CNE-2Z细胞中的表达最少（P＜0.01）。与对照组比较，实验组CNE-2Z细胞的增殖能力从第3天开始明显降低（P＜0.05），实验组CNE-2Z细胞迁移能力明显下降（P＜0.01）。与对照组比较，实验组CNE-2Z细胞中TFPI2在mRNA和蛋白水平的表达明显增加（P＜0.01），ERK信号通路蛋白p-ERK、MSK1、MNK和STAT3表达明显降低。结论:lncRNA LAMA5-AS1在鼻咽癌组织和细胞株中表达明显降低，高表达lncRNA LAMA5-AS1可通过上调TFPI2基因表达、下调ERK信号通路活性，抑制鼻咽癌CNE-2Z细胞的增殖和迁移能力。

lncRNA LAMA5-AS1 inhibits proliferation and migration of nasopharyngeal carcinoma cells by up-regulating TFPI2 expression

Objective:To detect the expression of LAMA5-AS1,a long non-coding RNA（lncRNA) in nasopharyngeal carcinoma tissues and cell lines,to observe the effect of lncRNA LAMA5-AS1 on the proliferation and migration of nasopharyngeal carcinoma cells and to explore its mechanism.Methods:Real-time quantitative polymerase chain reaction（qPCR) was used to detect the relative expression of lncRNA LAMA5-AS1 in 68 cases of nasopharyngeal carcinoma tissue and 62 cases of chronic nasopharyngitis tissue,nasopharyngeal carcinoma cell line,and immortalized nasopharyngeal epithelial cells.Cell lines with the least expression of lncRNA LAMA5-AS1 were selected and transfected with negative control plasmid（control group) or plasmids containing lncRNA LAMA5-AS1 sequence（experimental group).MTS method and cell scratch test were used to detect the effects of the overexpressing lncRNA LAMA5-AS1 on cell proliferation and migration ability.qPCR was used to detect the effect of the overexpressing lncRNA LAMA5-AS1 on mRNA expression of tissue factor pathway inhibitor 2（TFPI2) gene.Western blotting was used to detect the expression of TFPI2 protein and ERK signaling pathway proteins.Results:Compared with chronic nasopharyngitis tissues,lncRNA LAMA5-AS1 expression was significantly reduced in nasopharyngeal carcinoma tissues（P＜0.01).Compared with human immortalized nasopharyngeal epithelial cells,the expression of lncRNA LAMA5-AS1 was significantly reduced in nasopharyngeal carcinoma cell lines（P＜0.01),and the expression was lowest in CNE-2Z cells（P＜0.01).Compared with the control group,the proliferation capacity of CNE-2Z cells in the experimental group was significantly reduced from day 3（P＜0.05),and the migration capacity of CNE-2Z cells in the experimental group was significantly reduced（P＜0.01).Compared with the control group,the expression of TFPI2 in mRNA and protein levels of CNE-2Z cells in the experimental group increased significantly（P＜0.01),and the expression of ERK signaling pathway proteins p-ERK,MSK1,MNK,and STAT3 significantly decreased.Conclusion:The expression of lncRNA LAMA5-AS1 is significantly reduced in nasopharyngeal carcinoma tissues and cell lines.High expression of lncRNA LAMA5-AS1 can inhibit the proliferation and migration capacities of CNE-2Z cells by up-regulating TFPI2 gene expression and down-regulating ERK signaling pathway progession.