METTL14通过促进N^(6)甲基腺嘌呤(m^(6)A)Myc的表达促进宫颈癌细胞的增殖和迁移

METTL14 promotes the proliferation and migration of cervical cancer cells by up-regulating m^(6)A Myc expression

目的 探究甲基转移酶样蛋白14(METTL14)基因对宫颈癌细胞增殖和转移能力的影响及其可能的分子机制。方法使用基因表达数据集(GEO)数据库和宫颈癌组织芯片分析宫颈癌组织及癌旁组织的METTL14和Myc表达水平。采用RNA干扰技术(RNAi)沉默HeLa和SiHa宫颈癌细胞中METTL14的表达,实时荧光定量PCR(qPCR)验证沉默效果。敲低METTL14后,CCK-8实验、集落形成实验、 5-乙炔基-2′-脱氧尿苷(EdU)实验检测细胞增殖和集落形成能力,Transwell^(TM)实验检测细胞迁移能力。Western blot法测定敲低METTL14后METTL14和Myc的蛋白表达水平,甲基化RNA免疫共沉淀后实时定量PCR(MeRIP-qPCR)检测各组HeLa细胞中N^(6)甲基腺嘌呤(m^(6)A)Myc的表达水平。结果 GEO数据库和宫颈癌组织芯片染色结果显示,METTL14和Myc在宫颈癌组织中的表达显著高于宫颈癌癌旁组织,且METTL14高表达的宫颈癌患者生存期更短。沉默METTL14后能明显抑制宫颈癌HeLa和SiHa细胞的细胞活力、增殖和迁移能力,其作用机制可能与METTL14促进m^(6)A Myc的表达有关。结论 METTL14通过促进m^(6)A Myc的表达促进宫颈癌细胞的增殖和迁移。

Objective To investigate the effect of methyltransferase-like 14(METTL14) on the proliferation and metastasis of cervical cancer cells and its possible molecular mechanism. Methods The expression of METTL14 and Myc in cervical cancer tissues and normal tissues were analyzed using Gene Expression Omnibus(GEO) database and cervical cancer tissue microarray. The expression of METTL14 in HeLa and SiHa cells was silenced by small interfering RNA. After silencing the expression of METTL14 in cervical cancer HeLa and SiHa cells by RNA interference(RNAi), real-time quantitative PCR(qPCR) was used to verify the effect. CCK-8 assay, colony formation assay, 5-ethynyl-2′-deoxyuridine(EdU) assay were adopted to detect cell proliferation and colony forming ability. Transwell;assay was employed to evaluate cell migration ability. After knocking out METTL14, Western blot was used to detect the protein expression of METTL14 and Myc. Methylated RNA immunoprecipitation-qPCR(MeRIP-qPCR) was applied to observe the expression of m^(6)A Myc in HeLa cells in each group. Results GEO database analysis and cervical cancer tissue microarray staining showed that the expression of METTL14 and Myc in cervical cancer tissues was significantly higher than that in adjacent tissues, and the survival time of cervical cancer patients with high expression of METTL14 was shorter. Silencing METTL14 can significantly inhibit the cell viability, proliferation and migration of cervical cancer HeLa and SiHa cells, and its mechanism of action may be related to the up-regulation of the expression of m^(6)A Myc by METTL14. Conclusion METTL14 promotes the proliferation and migration of cervical cancer cells by up-regulating the expression of m^(6)A Myc.