miR-142a-3p Reduces Autophagy in TCMK-1 Cells and Enhances Pyroptosis by Targeting ATG16L1
10.13865/j.cnki.cjbmb.2025.06.1083
- VernacularTitle:miR-142a-3p通过靶向自噬相关16样蛋白1减轻TCMK-1细胞自噬而增强焦亡
- Author:
Xing ZHAO
1
;
Fei YU
;
Rui-Yang YUAN
;
Ya-Ru YANG
;
Jia-Yan LIU
;
Hai-Mai DING
;
Xue-Ming ZHANG
Author Information
1. 包头医学院基础医学与法医学院生物化学与分子生物学实验室,内蒙古包头 014040
- Publication Type:Journal Article
- Keywords:
microRNA-142a-3p(miR-142a-3p);
autophagy;
pyroptosis
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(7):1031-1039
- CountryChina
- Language:Chinese
-
Abstract:
The incidence rate of kidney diseases in China has always remained high.At present,the clinical treat-ment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of eco-nomical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and devel-opment of diseases.This study aims to explore the role and regulatory mechanism of miR-142a-3p in adriamycin(ADR)-induced renal tubular epithelial cell(TCMK-1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK-8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR-142a-3p and its target gene ATG16L1 mRNA levels were quantified using RT-qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR-142a-3p in TCMK-1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P＜0.0001).Western blotting results showed that the levels of p62(P＜0.001)and pyroptosis-related proteins(P＜0.001)were increased,while the protein levels of autophagy-related proteins were decreased(P＜0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph-agosomes between the ADR group and the autophagosome inhibitor group(3-MA group)(P＞0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR-142a-3p was inhibited by transfecting miR-142a-3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P＜0.001).Western blotting showed that the protein level of p62(P＜0.01)and pyropto-sis-related proteins(P＜0.01)were decreased,and the protein level of autophagy-related proteins was restored(P＜0.001).Flow cytometry results further indicated that cell autophagy was restored(P＜0.0001).In conclusion,ADR targets A TG1 6L1 through miR-142a-3p to reduce the autophagy level of TCMK-1,and simultaneously activates GSDMD-mediated pyroptosis.
