关键词: 宫颈癌, 瞬时受体电位阳离子通道亚家族M成员7, 甲基转移酶3, 铁死亡, N6-甲基腺苷

Abstract: Objective Methyltransferase 3(METTL3) mediated N6-methyladenosine (m6A) methylation modifica-tion of transient receptor potential cation channel subfamily M member 7(TRPM7) regulates ferroptosis and malignant progression in cervical cancer(CESC). Methods Totally 40 patients with cervical cancer were collected. Cervical cancer tissues and adjacent normal tissues(≥3 cm from the edge of the tumor tissue) were sampled at operation and then divided into experimental group and control group, respectively. RT-qPCR and Western blot were used to detect the differences in TRPM7 mRNA and protein expression between the two groups. TRPM7-interfering cell lines were constructed to investigate the effects of TRPM7 on CESC cells. Cell proliferation and apoptosis were assessed using 5-ethynyl-2'-deoxyuridine (EdU) assay and flow cytometry, respectively. Transwell chamber assays were employed to evaluate cell invasion and migration capabilities. The levels of ferroptosis in CESC cells were measured using kits for reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and Fe²⁺. Bioinformatics tools were utilized to predict methyltransferases associated with TRPM7. The interaction between METTL3 and TRPM7 was examined through RNA immunoprecipitation (RIP) and methylated RNA immunoprecipitation quantitative PCR (Me-RIP qPCR). The effect of METTL3 on the stability of TRPM7 expression was assessed using actinomycin D assay. Results TRPM7 was highly expressed in CESC tissue and cells. Knockdown of TRPM7 significantly inhibited cell proliferation, promoted cell apoptosis, suppressed cell migration and invasion capabilities, and enhanced ferroptosis levels (P＜0.05). Bioinformatics predictions suggested that METTL3 might act as a methyltransferase for TRPM7. Interference with METTL3 gene expression significantly reduced TRPM7 protein levels, decreased TRPM7 m6A modification levels, and impaired TRPM7 gene stability (P＜0.05). Conclusions METTL3 regulates CESC proliferation, apoptosis, migration, invasion, and ferroptosis by m6A methylation modification of the TRPM7 gene.

Key words: cervical squamous cell carcinoma, transient receptor potential cation channel subfamily M member 7, methyltransferase 3, ferroptosis, N6-methyladenosine