长链非编码RNA LINC00467促进非小细胞肺癌细胞的增殖
Effects of LncRNA-LINC00467 on cell proliferation of human non-small cell lung cancer

摘要：目的 研究长链非编码RNAs(Long non-coding RNAs, LncRNAs)LINC00467在非小细胞肺癌(non-small cell lung cancer, NSCLC)组织及细胞系中的表达及其对NSCLC细胞系功能的影响。方法 利用高通量基因表达[Gene Expression Omnibus (GEO)]数据库提取GSE19804和GSE18842数据集的生物信息学资料，分析LINC00467在NSCLC组织和正常肺组织中的表达差异；采用实时荧光定量PCR(qRT-PCR)检测收集的25对NSCLC癌组织及其癌旁组织以及NSCLC细胞系(A549、NCI-H266、NCI-H1299)和人正常支气管上皮细胞系HBE中LINC00467的表达；然后将NSCLC细胞系A549分别转染LINC00467 siRNA(si-LINC00467组) 和对照序列(si-NC组)，分别采用流式细胞技术、CCK8和细胞克隆形成实验检测两组细胞的增殖情况，qRT-PCR和Western blot实验检测两组细胞细胞周期相关分子G1/S-特异性周期蛋白-D1(CyclinD1)和细胞周期分子细胞周期素蛋白依赖性激酶6(CDK6)的表达。结果 GEO公共数据库分析显示，LINC00467在NSCLC组织中的表达高于正常肺组织(P<0.05)，与癌旁组织和上皮正常细胞相比较，NSCLC癌组织和细胞系中LINC00467的表达水平升高(P<0.05)；A549细胞中si-LINC00467组中LINC00467表达水平明显低于si-NC组(P<0.01)，且细胞活力和细胞克隆数均低于si-NC组，差异均有统计学意义(P<0.05)；相较于si-NC组，si-LINC00467组CyclinD1和CDK6的表达受到明显抑制(P<0.05)。结论 NSCLC组织及细胞系中LINC00467呈高表达，沉默LINC00467表达可明显抑制NSCLC恶性增殖的表型，可为NSCLC的靶向治疗提供潜在策略。
ABSTRACT: Objective To explore the expression level of long non-coding RNA LINC00467 in tissues and cells of colorectal cancer so as to study the effect of LINC00467 on the function of non-small cell lung cancer (NSCLC) cell line. Methods First, the expression of LINC00467 was analyzed with the Gene Expression Omnibus (GEO) database. Meanwhile, real-time quantitative PCR was applied to detect the expression of LINC00467 in 25 paired tumor tissues and corresponding adjacent normal tissues, normal lung epithelial HBE cell line and NSCLCA549, NCI-H266 and NCI-H1299 cell lines. Then, the specific small interfering RNA for LINC00467(si-LINC00467 group) or negative control sequence (si-NC group) were transfected into A549 cells. Flow cytometry, clone formation experiment and CCK8 assay were employed to detect the proliferation of the cells in the two groups. Finally, the expressions of CyclinD1 and CDK6 in the two groups were detected by qRT-PCR and Western blot. Results In the GEO databases GSE19804 and GSE18842, the expression of LINC00467 was upregulated in NSCLC; meanwhile, the level of LINC00467 expression in both NSCLC tissues and cells was higher than that in the adjacent normal tissues and normal cells (both P<0.05). Compared with si-NC group, there were significantly lower LINC00467 level (P<0.01), and cell activity and cell clone number in si-LINC00467 group (P<0.05). Meanwhile, compared with control group, the expressions of CyclinD1 and CDK6 in si-LINC00467 group were significantly decreased (P<0.05). Conclusion There is high expression of LINC00467 in NSCLC tissues and cells, and knock-down LINC00467 expression can inhibit the proliferation of NSCLC cells, which