N 6-methyladenosine-dependent pri-miR-17-92 mature activates AKT/mTOR pathway to promote endometrial cancer progression
10.3760/cma.j.cn.115807-20220907-00246
- VernacularTitle:N6-甲基腺苷依赖性pri-miR-17-92成熟激活AKT/mTOR途径促进子宫内膜癌进展
- Author:
Xiaoyan WANG
1
;
Hongyin CUI
;
Qingwen XIE
;
Xiaoqian ZHOU
;
Huanxin ZHONG
Author Information
1. 浙江大学医学院附属第二医院临平院区妇产科，杭州 311100
- Keywords:
Endometrial cancer;
N 6-methyladenosine;
METTL3;
miR-17-92;
AKT/mTOR
- From:
Chinese Journal of Endocrine Surgery
2022;16(6):698-702
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the role of N 6-methyladenosine (m6A) and its regulator METTL3 in the non-coding RNA of endometrial cancer.Methods:The expression levels of m6A and METTL3 were quantified in 20 paired carcinoma and adjacent clinical tissue samples from patients at from Jul. 2016 to Dec. 2020. HEC-1-A cell lines were constructed with METTL3 overexpression and knockdown. Western blot was used to detect the phosphorylation levels of key molecules in METTL3 and Akt/mTOR. The quantitative detection of mRNA levels were used qRT-PCR. The binding level of m6A to its receptor DGCR8 was determined by RNA immunoprecipitation.Results:The results of the m6A RNA methylation quantification kit showed that m6A (1.0±0.15) vs (1.7±0.34) ( P<0.01) and METTL3 levels were elevated in endometrial cancer cells, and METTL3 (1.0±0.13) vs (2.5±0.45) ( P<0.05) levels were elevated in endometrial cancer cells. Western blot and qRT-PCR detection of miR-17-92 cell clusters overexpressing METTL3, METTL3 overexpression significantly increased m6A modification on pri-miR-17-92 ( P<0.05) . Phosphorylation levels of AKT/mTOR pathway-related proteins were upregulated. In addition, RIP test results indicated that the binding of DGCR8 to pri-miR-17-92 was significantly facilitated. Conclusion:METTL3 modification of m6A facilitates the processing of pri-miR-1792 into the miR-17-92 clusters via m6A/DGCR8-dependent mechanism, which in turn activated the AKT/mTOR pathway.
