LncRNA CASC11在甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中的作用

Role of LncRNA CASC11 in the malignant transformation of 16HBE cells induced by glycidyl methacrylate

目的研究甲基丙烯酸环氧丙酯(glycidyl methacrylate,GMA)诱导人支气管上皮细胞(16HBE)恶性转化过程中趋势性上调差异表达LncRNA CASC11的作用。方法低浓度(8μg/mL)GMA重复染毒16HBE细胞后,收集染毒后第10代(P10)、20代(P20)和30代(P30)细胞及其同期二甲基亚砜(dimethyl sulfoxide,DMSO)对照组细胞,采用高通量LncRNA芯片筛选上述不同时期相关的差异表达LncRNAs,应用时间序列分析和生物信息学工具RPISeq筛选并预测特定LncRNA的可能作用靶点,实时荧光定量PCR(qPCR)和蛋白质印迹法分别检测特定LncRNA相对表达量及其靶蛋白含量。结果GMA诱导16HBE细胞恶性转化过程中特定差异表达LncRNA CASC11呈趋势性上调,与同期对照组细胞比较,在染毒后P10、P20、P30代细胞中分别下调1.64倍、上调2.01倍和上调2.66倍。与同期DMSO对照组相比,在染毒后P10、P20、P30代细胞中细胞周期蛋白依赖性激酶1(cyclin-dependent kinase 1,CDK1)及细胞周期蛋白B1、细胞周期蛋白B2表达水平均降低,与芯片结果一致。结论GMA诱导16HBE细胞恶性转化过程中相关差异表达LncRNA CASC11呈趋势性上调,其可能是通过抑制CDK1阻断或延迟细胞周期进程。

OBJECTIVE To investigate the effect of trending up-regulation LncRNA CASC11 which is differentially expressed during the malignant transformation of human bronchial epithelial cells(16 HBE)induced by glycidyl methacrylate(GMA).METHODS After 16 HBE cells were repeatedly exposed to low concentration GMA(8μg/mL),the 10 th,20 th and 30 th passage cells of the GMA group and the DMSO solvent control group were collected.High throughput LncRNA microarray were used to screen the differentially expressed LncRNAs at different stages.Short Time-series Expression Miner and bioinformatics tool RPISeq were used to screen and predict the potential targets of specific LncRNA.Real-time fluorescent quantitative PCR(qPCR)and Western blot were used to detect the relative expression of specific LncRNA and the content of its target protein respectively.RESULTS The specific differential expression of LncRNA CASC11 in the process of GMA-induced malignant transformation of 16 HBE cells showed a trend of up-regulation.Compared with the control cells in the same period,the P10,P20 and P30 generation cells were down-regulated by 1.64 times,up-regulated by 2.01 times,and up-regulated by 2.66 times,respectively.Western blot showed that the levels of cyclin-dependent kinase 1(CDK1),cyclinB1 and cyclinB2 in P10,P20 and P30 cells after exposure were lower than those in DMSO control group during the same period,which was consistent with the microarray results.CONCLUSION The differential expression of LncRNA CASC11 in the process of GMA-induced 16 HBE cell malignant transformation was up-regulated trendingly.It is speculated that it may block or delay cell cycle progression by inhibiting CDK1.