长链非编码RNA GIHCG对非小细胞肺癌恶性表型的影响

Effect of long non-coding RNA GIHCG on malignant phenotype of non-small cell lung cancer

目的探讨长链非编码RNA GIHCG(lncRNA GIHCG)对非小细胞肺癌(NSCLC)恶性表型的影响。方法使用qRT-PCR法分别检测50例NSCLC的肿瘤组织及其癌旁组织中lncRNA GIHCG及正常人支气管上皮细胞系E16HB,NSCLC细胞系A549、H1299、H1650、H2087中lncRNA GIHCG的表达。分别使用小干扰RNA(siRNA-1、siRNA-2)及对照转染A549和H1299细胞,实验设siRNA-1转染组、siRNA-2转染组、阴性对照组和空白对照组。各组转染48 h后采用CCK-8实验检测细胞增殖活性,Transwell实验检测细胞侵袭能力,流式细胞术检测各组细胞周期变化。结果qRT-PCR结果显示,lncRNA GIHCG在肺癌组织中的表达水平明显高于癌旁组织,同时非小细胞肺癌细胞系A549、H1299、H1650、H2087中lncRNA GIHCG的表达均高于16HBE细胞,差异有统计学意义(P<0.05)。转染48 h后,与空白对照组和阴性对照相比,siRNA-1组和siRNA-2组细胞存活率明显下降,穿膜细胞数明显减少,G0/G1期细胞比例明显增加,S期细胞减少,差异均有统计学意义,而阴性对照组和空白对照组上述指标差异均无统计学意义。结论lncRNA GIHCG高表达于NSCLC组织及细胞系中,沉默lncRNA GIHCG表达可明显抑制NSCLC恶性生物学表型。

Objective To investigate the effect of long non-coding RNA GIHCG(lncRNA GIHCG)on the malignant phenotype of non-small cell lung cancer(NSCLC).Methods Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of lncRNA GIHCG in the tumor tissues and the adjacent normal tissues of 50 patients with NSCLC,respectively.LncRNA GIHCG expression levels in human normal bronchial epithelial cell line 16HBE and NSCLC cell lines A549,H1299,H1650,and H2087 were also detected.Two small interfering RNAs(siRNA-1 and siRNA-2)were designed according to the lncRNA GIHCG sequence.Both the siRNAs were transfected respectively into A549 and H1299 cells.Then the cells were separately divided into four groups:siRNA-1 transfection group,siRNA-2 transfection group,the negative control group,and the blank control group.After transfection for 48 h,the cell proliferation activity was investigated with CCK8 array.Transwell assay was applied to detect the cell invasion ability and flow cytometry was used to detect the cell cycle changes in each group.All the data were statistically analyzed with SPSS 16.0.The measurement data conforming to the normal distribution were shown as mean±standard deviation(mean±SD),and the comparison between two groups was performed by independent sample t test.The one-way analysis of variance(ANOVA)was used to determine whether there were any statistically significant differences in comparing multiple groups and the least significant difference(LSD)was used to further analyze the difference between two groups after ANOVA test.Results The qRT-PCR results showed that the expression level of lncRNA GIHCG in the lung cancer tissues was significantly higher than that of the adjacent normal tissues,and the expression levels of lncRNA GIHCG in the NSCLC cell lines A549,H1299,H1650 and H2087 were all significantly higher than the expression level of GIHCG in 16HBE cells(P<0.05).After transfection for 48 h,compared with the blank control group and the negative control group,the survival rates of cells in the siRNA-1 transfection group and the siRNA-2 transfection group were significantly decreased.Correspondingly,the number of transmembrane cells was significantly reduced.The proportion of cells in G0/G1 phase was significantly increased,and the number of cells in the S phase was decreased,with statistically significant difference between them.However,the G2/M phase cells were the same proportion in different treatments.The difference between the negative control group and the blank control group had no statistical significant difference.Conclusion lncRNA GIHCG is highly expressed in the NSCLC tissues and the cell lines.Therefore,silencing the expression of lncRNA GIHCG can significantly inhibit the biological phenotype of malignant NSCLC.