甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中细胞凋亡相关lncRNA的筛选及功能研究

doi:10.3969/j.issn.1004-616x.2021.01.001

Abstract

Abstract: OBJECTIVE: To screen for apoptosis-related lncRNAs and to investigate their regulatory effects on glycidyl methacrylate(GMA)-induced malignant transformation of human bronchial epithelial (16HBE) cells. METHODS: The 16HBE cells in the logarithmic growth phase were exposed to dimethyl sulfoxide (DMSO) as the solvent control group, and to 8 μg/mL GMA as the treatment group. Cells were subcultured after repeated exposure 3 times for 72 hours each time. Subsequently, cells at the 10th, 20th and 30th generations were collected. Based on cell transformation phenotypes of the cultured cells, they were divided into pre-, middleand late-transformation stages. Flow cytometry was used to detect expression of apoptosis and apoptosis-related lncRNAs were screened from the lncRNA microarrays of the three transformation stages. Bioinformatics database was used to find the target genes of the differentially expressed lncRNAs. Real-time quantification PCR was used to detect the relative expression of differential lncRNAs. RESULTS: Compared with the control group of the same (10th) generation, the apoptosis levels of the GMA-treated cells were significantly increased. The apoptosis levels of the 20th generation GMA-treated cells were significantly lower than the same generation control group (P < 0.05) and were not significantly different from the control group of the 30th generation. The results from the lncRNA microarrays show that lncRNA CFLAR-AS1 was down-regulated 22%, up-regulated 244% and down-regulated 30% in the 10 th, 20th and 30th generations after GMA-treatments, suggesting that the target mRNA CFLAR was up-regulated 27%, down-regulated 30.6% and down-regulated 31%. The results from qPCR verification of lncRNA CFLAR-AS1 were consistent with the results of the lncRNA microarrays which were down-regulated 42%, up-regulated 442% and down-regulated 9% in the 10th, 20th and 30th generations, respectively. The difference between the 10th and the 20th generations was statistically significant (P < 0.05). CONCLUSION: In the process of malignant transformation of 16HBE from repeated exposure to GMA, the pre-transformation apoptosis ratio increased and the mid-transformation apoptosis ratio decreased. During this period, lncRNA CFLAR-AS1 played a role in inhibiting apoptosis but the mechanism for this function needs further study.

Key words: glycidyl methacrylate, human bronchial epithelial cells, malignant transformation, apoptosis, lncRNA

CLC Number:

R114

MA Shunpeng, WANG Quankai, WANG Miao, SONG Jiayang, WUHAN Baolier, XIE Guangyun, XU Jianning. Long non-coding RNA regulates apoptosis during glycidyl methacrylate-induced malignant transformation of 16HBE cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(1): 1-5,31.

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Long non-coding RNA regulates apoptosis during glycidyl methacrylate-induced malignant transformation of 16HBE cells

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