LncRNA GATA2-AS1在肾癌组织中的表达及其对肾癌细胞ACHN恶性生物学行为的影响

The expression of lncRNA GATA2-AS1 in renal cell carcinoma tissues and its effect on malignant biological behaviors of renal cancer cells ACHN

目的:检测长链非编码RNAs(lncRNAs)GATA2-AS1在肾细胞癌(RCC)组织及肾癌细胞株中的表达,并探讨其表达与患者临床病理资料之间的相关性。研究过表达GATA2-AS1对肾癌ACHN生物学行为及上皮间质转化(EMT)进程的影响。方法:应用实时荧光定量多聚核苷酸酶链式反应(RT-qPCR)检测GATA2-AS1在71例肾细胞癌组织和与其相对应癌旁正常组织以及肾癌细胞株(786-O、ACHN)中的表达。通过建立过表达载体pcDNA3.1-GATA2-AS1并转染ACHN细胞,应用RT-qPCR法检测转染效率。应用细胞增殖实验(MTS)、克隆形成实验检测GATA2-AS1过表达对ACHN细胞体外增殖能力的影响。应用划痕实验、Transwell小室侵袭实验分别检测GATA2-AS1过表达对ACHN细胞体外迁移能力及侵袭能力的影响。应用RT-qPCR法检测GATA2-AS1过表达对EMT进程中相关标志物(E-cadherin、N-cadherin及Vimentin)表达水平的影响。结果:GATA2-AS1在两株肾癌细胞系中相对表达量均明显下调(P<0.01),其中ACHN细胞的GATA2-AS1表达水平最低。GATA2-AS1在RCC组织中的相对表达量显著低于癌旁组织(P<0.01),GATA2-AS1在RCC组织中的表达水平与TNM分期及淋巴结转移相关(P<0.05或P<0.01)。成功构建过表达载体pcDNA3.1-GATA2-AS1并转染肾癌细胞ACHN,GATA2-AS1过表达组的相对表达量显著高于对照组(P<0.01)。GATA2-AS1过表达后可显著抑制ACHN细胞的在体外的增殖、迁移及侵袭能力(P<0.05或P<0.01)。同时E-cadherin表达水平上调,间充质标志物N-cadherin、Vimentin表达水平降低(P<0.05或P<0.01)。结论:GATA2-AS1在RCC组织及肾癌细胞株中低表达情况,与患者TNM分期及淋巴转移有关。GATA2-AS1过表达可以显著抑制肾癌细胞的体外增殖、迁移、侵袭能力及EMT的进程。

Objective:To detect the expression of long non-coding RNAs GATA2-AS1 in renal cell carcinoma(RCC)tissues and cell lines,and discuss the relationship between its expression and clinicopathological parameters of patients,as well as to study the effect of overexpression of GATA2-AS1 on the biological behavior and epithelial-mesenchymal transition(EMT)of renal cell carcinoma ACHN.Methods:Real-time quantitative polymerase chain reaction(RT-qPCR)was applied to detect the expression of GATA2-AS1 gene in 71 cases of renal cell carcinoma tissues and their corresponding adjacent normal tissues and renal cell carcinoma cell lines(786-O,ACHN).The overexpression vector pcDNA3.1-GATA2-AS1 was established and transfected into ACHN cells,and the transfection efficiency was detected by RT-qPCR.Cell proliferation assay(MTS)and cloning formation experiment were applied to detect the effect of GATA2-AS1 gene overexpression on the proliferation ability of ACHN cells in vitro.The effects of GATA2-AS1 overexpression on the migration and invasion ability of ACHN cells in vitro were detected by scratch experiment and Transwell chamber invasion experiment.The RT-qPCR method was applied to detect the effect of GATA2-AS1 overexpression levels of relevant markers(E-cadherin,N-cadherin and vimentin)in the EMT process.Results:The relative expression of GATA2-AS1 was significantly down-regulated in both renal cancer cell lines(P<0.01),and the expression level of GATA2-AS1 in ACHN cells was the lowest.The relative expression level of GATA2-AS1 in RCC tissue was significantly lower than that in adjacent tissues(P<0.01).The expression level of GATA2-AS1 in RCC tissue was correlated with TNM stage and lymph node metastasis(P<0.05 or P<0.01).The overexpression vector pcDNA3.1-GATA2-AS1 was successfully established and transfected into renal cell carcinoma cell line ACHN.The relative expression of GATA2-AS1 in the overexpression group was significantly higher than that in the control group(P<0.01).After GATA2-AS1 was overexpressed,it can significantly inhibit the proliferation,migration and invasion ability of ACHN cells in vitro.Meanwhile,the expression level of E-cadherin increased and the expression level of mesenchymal markers N-cadherin and Vimentin decreased.Conclusion:The low expression of GATA2-AS1 in RCC tissues and renal cancer cell lines was related to TNM staging and lymphatic metastasis in patients.Overexpression of GATA2-AS1 can significantly inhibit the proliferation,migration,invasion ability and EMT process of renal cell carcinoma cells in vitro.