非小细胞肺癌组织中长链非编码RNA-THOR的表达意义及生物学作用

Expression of RNA-THOR in tissues of non-small cell lung cancer and its pathological value

目的探讨非小细胞肺癌(non-small-cell lung cancer,NSCLC)癌组织和癌旁正常组织中睾丸相关高度保守的致癌长链非编码RNA(testis-associated highly-conserved oncogenic long non-coding RNA,THOR)的表达及其与NSCLC临床病理特征关系;并初步探讨其作用和机制。方法选取79例诊断确认的NSCLC患者癌组织和其对应的癌旁正常组织(距离肿瘤组织>5cm),采用实时荧光PCR方法,比较THOR在二者中的表达差异情况,并通过肿瘤直径、组织学类型、TMN分期、分化程度等临床病理特征因素分析THOR表达量与患者预后的相关性。瞬时转染siRNA沉默A549细胞中THOR的表达,通过CCK-8法检测对细胞增殖的影响,Western blot法检测,对细胞周期蛋白cyclin D1及凋亡相关蛋白Bcl2、BAX、cleaved-caspase-3的影响。结果实时荧光RT-PCR结果显示NSCLC癌组织中的THOR表达量显著高于对应的癌旁正常组织,差异具有统计学意义(P<0.05)。THOR的表达量与肿瘤直径、TMN分期及分化程度因素有相关性(P<0.05)。THOR高表达的NSCLC患者生存率低于THOR低表达的NSCLC患者。THOR表沉默后,A549细胞增殖受到抑制,cyclin D1及Bcl2表达降低,BAX、cleaved-caspase-3蛋白表达增多。结论THOR在NSCLC癌组织中的高表达,与NSCLC发生、发展及临床病理特征关系密切;并促进A549细胞增殖,并拮抗细胞凋亡,其可能的机制与调控周期蛋白cyclin D1表达有关。

Objective To explore the expression of testis-associated highly-conserved oncogenic long non-coding RNA(THOR)in human non-small cell lung cancer(NSCLC)and its correlation with clinic pathological features of NSCLC patients,and to preliminary discuss its role and mechanism.Methods A total number of 71 tissue specimens from the patients with NSCLC were enrolled as the experimental group,and the normal tissues 5cm away from tumor of the same patient were enrolled as the control group simultaneously.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the THOR expression level in the two groups.Through tumor diameter,TMN stage,lymph node metastasis and other clinic pathological factors,the relationship between the expression of THOR and the prognosis of patients was analyzed.Transient transfection of siRNA silenced the expression of THOR in A549 cells,CCK-8 was used to detecte the effect on cell proliferation,and apoptosis-related proteins Bcl2,BAX,and cleaved-caspase-3 were detected by Western blot.Results The results showed that the THOR expression level in NSCLC tissues was higher than that in adjacent normal tissues,and the difference was statistically(P<0.05).In addition,the expression of THOR was related to the degree of tumor diameter,TMN stage and lymph node metastasis(P<0.05).The survival rate of NSCLC patients with high expression of THOR was lower than those NSCLC patients with low expression.After silenced THOR expression,the proliferation of A549 cells was inhibited,and the protein expression of cyclin D1 and Bcl2 were decreased and the protein expression of BAX and cleaved-caspase-3 were increased.Conclusion There is a closely relationship between the high expression of THOR with the occurrence,development and clinic pathological characteristics of NSCLC.THOR promotes the proliferation of A549 cells and antagonizes apoptosis.The possible mechanism is related to the regulation of the expression of cyclin D1.