昆明医科大学学报2024，Vol.45Issue(6)：22-35,14.DOI:10.12259/j.issn.2095-610X.S20240604
miR-205-5p靶向ERBB3调控PI3K/AKT/mTOR通路抑制血管生成在痔疮中的分子机制
The Molecular Mechanism of miR-205-5p Targeting ERBB3 to Regulate PI3K/AKT/mTOR Pathway and Inhibit Angiogenesis in Hemorrhoids
摘要
Abstract
Objective To explore the molecular mechanism of miR-205-5p targeting ERBB3 to regulate PI3K/AKT/mTOR pathway and inhibit angiogenesis in hemorrhoids.Methods The hemorrhoidal nucleus and normal perianal tissues of 12 patients from July 2021 to June 2022 were collected,the expression of miR-205-5p and ERBB3 in the samples was detected by qRT-PCR,and the targeting relationship was verified by dual-luciferase.miR-205-5p mimic,pcDNA-ERBB3,miR-205-5p inhibitor,sh-ERBB3,oe-ERBB3 and negative control plasmids were respectively or co-transfected into hemorrhoidal model rats and HUVEC cells.The pathological changes and apoptosis of perianal tissue in each group were observed by HE and TUNEL staining,and the localization of expression of ERBB3 and VEGFR2 proteins were detected by immunohistochemistry,and the expression levels of ERBB3,VEGFR2,Cyclin D1,cleaved-caspase 3,Bax,Bcl-2 and PI3K/AKT/mTOR pathway related proteins were detected by Western blot.The proliferation,migration,invasion,apoptosis and angiogenesis of HUVEC were detected by MTT,wound healing,transwell,flow cytometry and angiogenesis assay.Results MiR-205-5p was lowly expressed and ERBB3 was highly expressed in clinical samples of hemorrhoids(P<0.001),and miR-205-5p targeted the 3'UTR of ERBB3(WT)(P<0.001).The expression of ERBB3 in hemorrhoidal rats and HUVEC cells in miR-205-5p mimic group was significantly decreased(P<0.01,P<0.001),and the levels of VEGFR2(P<0.001),Cyclin D1(P<0.01),Bcl-2(P<0.001),p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR(P<0.001)were significantly down-regulated,the levels of Cleaved-caspase 3 and Bax were significantly up-regulated(P<0.01),the perianal histopathology of rats was improved,the proliferation,migration and invasion activities of HUVEC cells were reduced(P<0.001),the apoptosis rate was increased(P<0.001),and the number and branches of angiogenesis were reduced(P<0.001),and these effects were eventually reversed by pcDNA-ERBB3(P<0.001).Conclusion MiR-205-5p can reduce angiogenesis and relieve hemorrhoids by targeting the inhibition of ERBB3 expression,down-regulating the activity of PI3K/AKT/mTOR pathway,inhibiting cell proliferation,migration and invasion,and promoting apoptosis.关键词
痔疮/miR-205-5p/ERBB3/PI3K/AKT/mTOR通路/血管生成Key words
Hemorrhoids/miR-205-5p/ERBB3/PI3K/AKT/mTOR pathway/Angiogenesis分类
医药卫生引用本文复制引用
李志霄,郑霞,李春玲,刘庆圣,张衡..miR-205-5p靶向ERBB3调控PI3K/AKT/mTOR通路抑制血管生成在痔疮中的分子机制[J].昆明医科大学学报,2024,45(6):22-35,14.基金项目
云南省教育厅科学研究基金资助项目(2023Y0436) （2023Y0436）
云南省科技厅科技计划基金资助项目(202101AZ070001-245). （202101AZ070001-245）
