m6a 甲基转移酶 METTL3 调控 KAI1/CD82 表达介导垂体 神经内分泌肿瘤细胞生物学行为的机制研究.

Objective To study the mechanism of m6a methyltransferase METL3 mediated proliferation, migration and invasion of pituitary neuroendocrine tumor cells by regulating the expression of KAI1/CD82. Methods The expression levels of METTL3 and KAI1/CD82 in rat pituitary cells, rat pituitary tumor cell lines GH3, and MMQ were determined by qPCR and Western blot assay. GH3 cells were cultured in vitro and randomly divided into the control group, the METL3 overexpression plasmid group, the METL3 empty plasmid group, the METL3 siRNA group and the METL3 siRNA negative control group. After grouping and transfection, the expression levels of METL3 and KAI1/CD82 of each cell group were detected by qPCR and Western blot assay. The cell viability of each group was detected by CCK-8 experiment. The cell migration and invasion in each group were detected by cell scratch and Transwell invasion experiment. The methylation modification of KAI1/CD82 m6A in each group was detected by methylated RNA immunoprecipitation (MeRIP) experiment. Results Compared with rat pituitary cells, the METL3 protein and mRNA expression level in rat pituitary tumor cell lines GH3 and MMQ were significantly increased (P＜0.05). KAI1/CD82 protein and mRNA expression levels were significantly reduced (P＜0.05). There were no significant differences in cell indicators between the control group, the METTL3 empty plasmid group and the METTL3 siRNA negative control group (P＞0.05). Compared with the control group and the METTL3 empty plasmid group, the cell viability, migration distance, number of invaded cells, METL3 protein and mRNA expression level, KAI1/CD82 m6A methylation level increased in the METL3 overexpression plasmid group (P＜0.05), and the KAI1/ CD82 protein and mRNA expression levels decreased (P＜0.05). Compared with the control group and the METTL3 siRNA negative control group, the cell viability, migration distance, number of invaded cells, METTL3 protein and mRNA expression level and KAI1/CD82 m6A methylation level decreased in the METTL3 siRNA group (P＜0.05), and KAI1/CD82 protein and mRNA expression levels increased (P＜0.05). Conclusion Down-regulating the expression of METTL3 can reduce the level of KAI1/CD82 m6A methylation, promote the transcriptional expression of KAI1/CD82 mRNA, reduce the proliferation, invasion and migration of pituitary tumor cells, and inhibit the occurrence and development of pituitary tumors. [ABSTRACT FROM AUTHOR]