lncRNA LOC440173在非小细胞肺癌组织中的表达及其对癌细胞恶性生物学行为的影响

Expression of lncRNA LOC440173 in non-small cell lung cancer tissues and its influence on the maligant biological behaviors of cancer cells

目的:检测lncRNA LOC440173在NSCLC组织和细胞中的表达及探讨其对癌细胞恶性生物学行为的影响。方法:选取河北医科大学第四医院生物标本库中2014至2017年手术切除的72例NSCLC患者的癌及癌旁组织标本,应用qPCR法检测NSCLC组织和癌旁组织中,以及6种NSCLC细胞株(H520、H358、A549、HCC827、H1703和H1299)中LOC440173的表达水平;构建LOC440173的敲低及过表达载体,分别转染H520和H1703细胞,应用MTS、克隆形成及Transwell小室迁移和侵袭实验分别检测敲低及过表达LOC440173对NSCLC细胞增殖、迁移及侵袭能力的影响,qPCR法检测LOC440173对于EMT过程相关标志物(E-cadherin、N-cadherin及vimentin)mRNA表达水平的影响,WB法检测其对E-cadherin、N-cadherin蛋白表达的影响。结果:LOC440173在NSCLC组织中的表达明显高于癌旁组织(P<0.01),并与淋巴结转移、组织学分化程度、TNM分期和肿瘤大小有关联(P<0.05或P<0.01)。敲低LOC440173可以抑制H520细胞的体外增殖、迁移和侵袭(P<0.05或P<0.01),过表达LOC440173可显著促进H1703细胞的增殖、迁移和侵袭(P<0.05或P<0.01)。在转录水平上,敲低LOC440173后,E-cadherin的表达水平升高,间充质相关标志物N-cadherin、vimentin的表达水平降低(P<0.05或P<0.01);而过表达LOC440173后,E-cadherin的表达水平降低,间充质相关标志物N-cadherin、vimentin的表达水平升高(P<0.05或P<0.01)。在转录后水平上,LOC440173负向调节E-cadherin蛋白的表达、正向调节N-cadherin的蛋白表达(均P<0.05)。结论:LOC440173在NSCLC组织中的异常高表达可能与NSCLC的发生发展有关,LOC440173可显著提高NCSCL细胞的体外增殖、迁移、侵袭能力,且其作用机制可能与调控EMT相关基因表达有关。

Objective:To detect the expression of lncRNA LOC440173 in NSCLC tissues and cells and to explore its influence on the maligant biological behavior of cancer cells.Methods:The cancer and para-cancerous tissues removed from 72 patients with NSCLC who were surgically during 2014 to 2017 in the biological specimen library of the Fourth Hospital of Hebei Medical University were selected.qPCR method was applied to detect the expression of LOC440173 in NSCLC tissues and the corresponding para-cancerous tissues as well as in six NSCLC cell lines(H520,H358,A549,HCC827,H1703 and H1299).The vectors used for LOC440173 knockdown or overexpression were constructed and transfected into H520 and H1703 cells,respectively.The effects of LOC440173 knockdown or overexpression on proliferation,migration,and invasion of lung cancer cells were examined by MTS,Clone formation,Transwell migration and invasion assays,respectively.qPCR method was used to detect the regulatory effect of LOC440173 on mRNA expression of EMT-related markers(E-cadherin,N-cadherin,and vimentin),WB method was employed to observe the protein expression of E-cadherin and N-cadherin.Results:The expression of LOC440173 in NSCLC tissues was significantly higher than that in corresponding para-cancerous tissues(P<0.01),and was correlated with lymph node metastasis,histological grade,TNM stage,and tumor size(P<0.05 or P<0.01).LOC440173 gene knockdown could inhibit the in vitro proliferation,invasion and migration of H520 cells(P<0.05 or P<0.01).Overexpression of LOC440173 gene significantly promoted the in vitro proliferation,migration,and invasion of H1703 cells(P<0.05 or P<0.01).At the transcriptional level,knockdown of LOC440173 was found to promote the expression level of E-cadherin and inhibit the expression level of N-cadherin and vimentin(P<0.05 or P<0.01),while overexpression of LOC440173 displayed the opposite results(P<0.05 or P<0.01).At the post-transcriptional level,LOC440173 negatively regulated protein expression of E-cadherin and positively modulated protein expression of N-cadherin(P<0.05).Conclusion:The abnormal high expression of LOC440173 may be related to the occurrence and development of NSCLC.LOC440173 can significantly improve the in vitro proliferation,migration and invasion of NSCLC cells,and the mechanism may be related to the regulation of EMT-related genes.