GMA诱导16HBE恶性转化细胞中LncRNA CTBP1-AS1的表达及意义

Expression and Significance of LncRNA CTBP1-AS1 in Glycidyl Methacrylate-Induced Malignantly Transformed 16HBE Cells

目的探讨LncRNA CTBP1-AS1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/m L GMA诱导的第30代16HBE恶性细胞及同代龄DMSO对照组细胞,应用高通量LncRNA芯片比较两组样本表达谱差异。通过差异倍数、邻近编码基因信息分析等策略筛选出16HBE恶性转化细胞LncRNA CTBP1-AS1及其最可能的蛋白质编码基因CTBP1,采用全基因组表达谱芯片和实时荧光定量PCR(qPCR)验证LncRNA CTBP1-AS1和CTBP1 mRNA的相对表达量。结果LncRNA芯片结果显示,与同代龄DMSO对照组相比,GMA诱导的16HBE恶性转化细胞中CTBP1-AS1上调2.20倍,CTBP1 mRNA上调1.26倍;qPCR结果显示,与同代龄DMSO对照组(26.74±0.23)×10^-5相比,GMA诱导的16HBE恶性转化细胞(91.70±0.70)×10^-5中LncRNA CTBP1-AS1表达明显上调(P<0.05),并与LncRNA芯片结果一致。结论GMA可以诱导16HBE细胞LncRNA CTBP1-AS1表达上调,LncRNA CTBP1-AS1可作为GMA诱导的16HBE恶性转化细胞中相关特异分子标志物。

Objective To investigate the expression and significance of long non-coding RNA(LncRNA)CTBP1-AS1(CTBP1 antisense RNA 1)in glycidyl methacrylate(GMA)-induced malignantly transformed 16 HBE cells.Methods The 30 th generation of malignantly transformed 16 HBE cells(induced by 8μg/m L GMA)and the same generation of control cells(treated with DMSO)were harvested.High-throughput LncRNA microarray analysis was used to measure the difference in the expression profile of LncRNA between the two groups.Strategies such as fold change analysis and neighboring gene analysis were used to preliminarily identify LncRNA CTBP1-AS1 and the most likely protein-coding gene CTBP1 in the malignantly transformed 16 HBE cells.Whole genome expression profile microarray and quantitative real-time PCR(qPCR)were used to verify the relative expression of LncRNA CTBP1-AS1 and CTBP1 mRNA.Results The result of LncRNA microarray analysis showed that the CTBP1-AS1 and CTBP1 mRNA in the GMA group were upregulated 2.20-fold and 1.26-fold,respectively,compared with those in the DMSO group;the result of q PCR showed that the expression of LncRNA CTBP1-AS1 in the GMA group was significantly higher than that in the DMSO group[(91.70±0.70)×10^-5 vs(26.74±0.23)×10^-5,P<0.05],which was consistent with the result of LncRNA microarray analysis.Conclusion GMA can induce the upregulation of LncRNA CTBP1-AS1 expression in 16 HBE cells,and LncRNA CTBP1-AS1 can be considered as a specific molecular marker in GMA-induced malignantly transformed 16 HBE cells.