肿瘤酸性微环境促进肾癌细胞株769-P体外侵袭及其lncRNA表达谱分析

Tumor acidic microenvironment promotes invasion of renal carcinoma 769-P cell line and analysis of the LncRNA expression profile

目的探索肿瘤胞外酸性微环境对肾癌细胞769-P侵袭性的影响,并分析其长链非编码RNA(lncRNA)表达谱。方法使用pH6.6酸性培养基体外模拟肿瘤酸性微环境,将肾癌细胞769-P于酸性培养基中培养,作用时间梯为0h、0.5h、1h、2h、4h、8h,并得到相应培养条件下的肾癌细胞。使用荧光定量PCR检测Snail基因及其下游E-cadherin基因的表达;通过transwell肿瘤细胞体外侵袭实验分析酸性条件下培养的肾癌细胞其侵袭能力的改变;最后通过lncRNA表达谱芯片分析酸性培养基作用后,肾癌细胞769-P的lncRNA表达谱变化。结果 pH 6.6酸性培养液培养4h后肾癌细胞769-P的Snail基因表达水平较对照组即pH6.6酸性培养液培养0h组显著上调(P<0.05),下游E-cadherin基因表达与其呈负相关,即pH6.6酸性培养液培养4h组显著下调(P<0.05);transwell肿瘤细胞体外侵袭实验证实,经pH6.6酸性培养液培养4h,肾癌细胞769-P的侵袭性较对照组显著提高(P<0.05);lncRNA表达谱芯片筛选结果显示,与对照组相比,经pH6.6酸性培养液培养4h后,肾癌细胞769-P中BC040587、KCNQ1DN、PCAT-43表达水平上调(P<0.05),CTBP1-AS、MEG3表达水平下调(P<0.05)。结论肿瘤胞外酸性微环境增强肾癌细胞769-P的侵袭能力,其作用机制可能与lncRNA BC040587、KCNQ1DN、PCAT-43、CTBP1-AS、MEG3的异常表达有关。

Objective To explore the effects of tumor acid microenvironment on the invasion of 769-P cell line and to analyze the LncRNA expression profile.Methods 769-P cells were cultured in pH6.6 cell culture medium for 0 h,0.5 h,1 h,2 h,4 h,and 8 h respectively.The expressions of Snail gene and E-cadherin were examined with fluorescent quantitative PCR.The invasion of 769-P cells was detected with Transwell assay.The LncRNA expression profile was analyzed with LncRNA expression profile chip.Results Compared with the 0 h 769-P cells,the 4 h 769-P cells showed significantly upregulated Snail gene while downregulated E-cadherin（both P〈0.05）;the invasion was remarkably enhanced（P〈0.05）;the expression levels of BC040587,KCNQ1DN and PCAT-43 were upregulated,while the expression levels of CTBP1-AS and MEG3 were downregulated（all P〈0.05）.Conclusions Tumor acidic microenvironment can enhance the invasion of 769-P cells.The mechanism may be related to the abnormal expressions of BC040587,KCNQ1DN,PCAT-43,CTBP1-AS and MEG3.