N6-甲基腺苷调控子宫内膜癌增殖的机制研究

Mechanism of N6-methyladenosine in regulating the proliferation of endometrial cancer

目的 通过干扰N6-甲基腺苷(M6A)的甲基化酶和去甲基化酶的表达,探讨M6A调控子宫内膜癌增殖的具体机制。方法 分别在人子宫内膜癌细胞系HEC-1-A和Ishikawa中过表达甲基转移酶样蛋白3(METTL3)敲降脂肪和肥胖相关蛋白(FTO),并使用环亮氨酸培养细胞,将其分为空白对照组、METTL3过表达组、METTL3过表达+环亮氨酸处理组、环亮氨酸处理组、敲降对照组、FTO敲降组,进行细胞计数-8实验研究各组细胞的增殖情况。结果 敲降FTO后,HEC-1-A和Ishikawa细胞增殖能力减弱。过表达METTL3后,HEC-1-A和Ishikawa细胞增殖能力减弱。环亮氨酸处理后,HEC-1-A和Ishikawa细胞增殖能力增强。METTL3过表达后,再进行环亮氨酸处理,HEC-1-A细胞增殖能力相较METTL3过表达组增强。结论 FTO和METTL3可通过改变M6A的水平调控子宫内膜癌细胞的增殖。

Objective To investigate the mechanism of N6-Methyladenosine(M6 A) regulating the proliferation of endometrial cancer by interfering the expression of M6 A methylase and demethylase. Methods Human endometrial cancer cell lines HEC-1-A and Ishikawa cells were overexpressed with methyltransferase-like 3(METTL3)and fat-mass and obesity associated protein(FTO)knockdown, respectively, and were cultured with cycloleucine. They were divided into the blank control group, the METTL3 overexpression group, the METTL3 overexpression + cycloleucine treatment group, the cycloleucine treatment group, the siRNA-negative group and the FTO-siRNA group. Cell counting kit-8 experiment was performed to study cell proliferation in each group. Results After knockdown of FTO, the proliferation ability of HEC-1-A and Ishikawa cells was decreased. After overexpression of METTL3, the proliferation ability of HEC-1-A and Ishikawa cells was decreased. After cycloleucine treatment, the proliferation ability of HEC-1-A and Ishikawa cells was increased. After METTL3 overexpression and cycloleucine treatment, the cell proliferation ability of HEC-1-A cells was enhanced compared with METTL3 overexpression group. Conclusion FTO and METTL3 can regulate the proliferation of endometrial cancer cells by changing the level of M6 A.