Mechanism of HIF1α regulating integrin protein αν expression by activating ALKBH5
10.3760/cma.j.cn101441-20220130-00051
- VernacularTitle:HIF1α通过启动ALKBH5调控整合素蛋白αν表达的机制研究
- Author:
Liying ZHOU
1
;
Yudi ZHANG
1
;
Li WANG
1
;
Shuyu WANG
1
Author Information
1. 首都医科大学附属北京妇产医院 北京妇幼保健院生殖医学科，北京 100026
- Publication Type:Journal Article
- Keywords:
Endometrial receptivity;
Hypoxia inducible factor-1α;
ALKBH5;
N6-methyladenosine methylation;
Integrin αν protein
- From:
Chinese Journal of Reproduction and Contraception
2023;43(4):371-380
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the intrinsic mechanism of hypoxia inducible factor 1α (HIF1α) initiating N6-methyladenosine (m6A) demethylase ALKBH5 expression to improve the expression of integrin αν (ITGAV), a molecular marker of endometrial receptivity.Methods:Human endometrial cells Ishikawa were used in this study. The expression of ALKBH5 was detected by real time quantity PCR (RT-PCR) and Western blotting under hypoxia and HIF1α interference conditions. After overexpression or interference ALKBH5, RT-PCR and Western blotting were used to detect ITGAV expression; Luciferase reporter gene system was used to detect miR-136-5p targeting ITGAV mRNA expression; RNA pull-down analysis and m6A methylation were used to detect the competitive binding of ALKBH5 and miR-136-5p to the methylation site sequence on ITGAV mRNA. Results:HIF1α promoted ALKBH5 protein expression in a time-dependent manner under hypoxic conditions in Ishikawa cells. Overexpression of ALKBH5 enhanced the expression of ITGAV and cell proliferation ( P<0.001, P<0.001), while knockout of ALKBH5 inhibited the expression of ITGAV and cell proliferation ( P=0.001, P=0.019). ITGAV mRNA ( P=0.007) and protein expression level ( P=0.015) were significantly down-regulated by miR-136-5p. And the location of mRNA sequence by miR-136-5p targeted overlapped with the methylation site recognized by ALKBH5, furthermore competed in regulating ITGAV mRNA. Conclusion:HIF1α regulates the expression of ITGAV and improves endometrial receptivity by regulating the expression of ALKBH5 and competitively binding to the specific sequence on ITGAV mRNA targeted by miR-136-5p.
