长链非编码RNA AL139147.1在胃癌中的表达及对MKN-45胃癌细胞生长的影响

Expression of long non-coding RNA AL139147.1 in gastric cancer and its effect on MKN-45 gastric cancer cell growth

目的:探讨长链非编码RNA(long non-coding RNA,LncRNA)AL139147.1在胃癌中的表达及其对胃癌细胞增殖和生长周期的影响。方法:利用癌症基因组图谱(TCGA)和基因型—组织表达数据库(GTEx)中的高通量测序数据,对AL139147.1表达及其与临床资料的相关性进行分析。按照AL139147.1表达高低对胃癌样本分组进行生存分析。通过慢病毒介导RNA干扰,沉默MKN-45细胞株中AL139147.1表达,筛选合适的shRNA;将MKN-45细胞分为空白对照组、阴性对照组、shRNA#3组,采用MTT比色法及流式细胞术检测细胞增殖和周期变化。结果:AL139147.1在胃癌组织中表达明显高于癌旁组织和正常胃黏膜组织(P均<0.05)。与AL139147.1低表达患者相比,高表达患者总体生存及无病生存较差(P均<0.05)。AL139147.1表达与T分期、TNM分期以及肿瘤分化程度相关(P均<0.05)。与空白对照组和阴性对照组相比,shRNA#3组MKN-45细胞增殖速度明显减缓,S期细胞比例明显降低,G2/M期细胞比例明显增高(P均<0.05)。结论:长链非编码RNA AL139147.1在胃癌中呈高表达,其低表达可减缓胃癌细胞生长增殖,并使细胞周期停滞于G2/M期。

Objective:To investigate the expression of long non-coding RNA(LncRNA)AL139147.1 in gastric cancer as well as its effect on proliferation and cell cycle.Methods:High-throughput sequencing materials from the Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)were used to analyze the AL139147.1 expression and its correlation with clinical parameters.Gastric cancer samples were grouped according to the expression level of AL139147.1 and survival analysis was then conducted.Subsequently,lentiviral mediated RNA interference was applied for the silence of AL139147.1 expression in MKN-45 cell line to select the optimal shRNA;then MKN-45 cells were divided into blank group,negative control group and shRNA#3 group.MTT colorimetry and flow cytometry were implemented to detect cell proliferation and cycle alterations.Results:The expression of AL139147.1 in gastric cancer tissues was significantly higher than that in adjacent tissues and normal gastric mucosa(all P<0.05).Patients with higher expression of AL139147.1 had poorer overall survival and disease-free survival(both P<0.05).The expression of AL139147.1 was significantly correlated with T stage,TNM stage and tumor differentiation(all P<0.05).Compared with the blank control group and the negative control group,the proliferation rate of MKN-45 cells in the shRNA#3 group was remarkably decreased,the ratio of S phase cells was markedly reduced,and the ratio of G2/M phase cells was greatly increased(all P<0.05).Conclusion:Long non-coding RNA AL139147.1 is highly expressed in gastric cancer,and its declined expression could slow down the growth and proliferation of cancer cells,and arrest the cell cycle in G2/M phase.