目的:探讨长链非编码RNA PCA3(LncRNA PCA3)在甲基丙烯酸环氧丙酯(GMA)诱导的16HBE恶性转化细胞中的表达水平及意义。方法:收获经8μg/mL GMA诱导的第30代16HBE恶性转化细胞及同代龄DMSO溶剂对照组细胞,应用高通量LncRNA芯片比较两组样本表达谱的差异,通过差异倍数、邻近编码基因信息分析等策略初步筛选出16HBE恶性转化细胞中LncRNA PCA3及其最可能的相关蛋白编码基因PRUNE2,采用实时荧光定量PCR(qPCR)和全基因组表达谱芯片分析LncRNA PCA3和PRUNE2的表达量,并与同代龄DMSO对照组细胞比较。结果:LncRNA芯片结果显示,与同代龄DMSO组相比,GMA诱导的16HBE恶性转化细胞中LncRNA PCA3上调7.17倍,PRUNE2下调2.54倍;qPCR结果显示,与同代龄DMSO组[(1.36±0.44)×10~(-5)]相比,GMA诱导的恶性转化细胞[(2.67±0.63)×10~(-5)]中LncRNA PCA3表达上调(P〈0.05);全基因组表达谱芯片显示,16HBE恶性转化细胞的PRUNE2表达量(10.95)较DMSO组(19.46)明显下调,与LncRNA芯片结果一致。结论:LncRNA PCA3可作为GMA诱导的16HBE恶性转化细胞中相关特异分子标志之一。
OBJECTIVE:To investigate the induction of expression of long non-coding RNA PCA3(LncRNA PCA3) by glycidyl methacrylate(GMA) in malignantly transformed 16 HBE cells.METHODS:Malignantly transformed 16 HBE cells(treated with 8 μg/mL GMA) and solvent control cells(DMSO) of the 30~(th) generations were harvested.High throughput LncRNA microarray was used to detect differences in expression profile of LncRNAs among the two groups of cells.Changes in LncRNAs were screened through strategies such as fold change and neighboring genes analysis.Real-time fluorescence quantitative PCR(qPCR) and gene chip were applied to measure the expression levels of LncRNA PCA3 and PRUNE2,respectively.RESULTS:Based on the result of LncRNA microarrays,LncRNA PCA3 in GMAtreated cells was up-regulated by 7.17 folds while PRUNE2 was down-regulated by 2.54 folds.QPCR showed that the expression of LncRNA PCA3[(2.67±0.63) X 10~5]in GMA-treated cells was significantly higher than that in the solvent control cells[(1.36±0.44)×10~(-5)](P〈0.05).On the other hand,the gene chip analyses showed that expression of PRUNE2(10.95) was reduced compared to the solvent control cells(19.46).CONCLUSION:LncRNA PCA3 expression can be considered as one of the stable and specific biomarkers involved in GMA- malignant transformation of 16 HBE cells.