Effect and mechanism of circZNF652/miR-140-3p/HMGB1 pathway on cell proliferation and migration in prostate cancer
10.3969/j.issn.1671-8348.2025.02.004
- VernacularTitle:circZNF652/miR-140-3p/HMGB1通路对前列腺癌细胞增殖和迁移的影响及机制
- Author:
Hua JIANG
1
;
He ZHANG
;
Songsong JIANG
Author Information
1. 遵义医科大学第五附属(珠海)医院/珠海市第六人民医院泌尿外科,广东珠海 519100
- Keywords:
circZNF652;
miR-140-3p;
HMGB1;
prostate cancer
- From:
Chongqing Medicine
2025;54(2):303-312,318
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression of circZNF652 in prostatic cancer(PCa)tissues and PCa cell lines,and its impact and possible mechanism on proliferation and migration.Methods Differential ex-pression of circRNAs in plasma tissues from PCa patients and benign prostatic hyperplasia(BPH)patients were detected by using gene chip technology.The expression levels of circZNF652 in PCa cell lines(22RV1,LNCaP,DU145,PC3)and normal prostate epithelial cells were measured by quantitative real-time PCR(qPCR).The correlation between different circZNF652 expression levels with the overall survival(OS)time and clinicopathological features of PCa patient were analyzed.The effects of interfering circZNF652 on PCa cell proliferation,colony formation,invasion and migration ability were assessed through CCK-8 assay,colony formation assay,Transwell assay,wound healing assay and EdU cell proliferation assay.The online database TargetScan was used to predict the binding sites between circZNF652 and miR-140-3p,and the dual-luciferase reporter gene assay and RNA pull-down assay confirmed the interaction of circZNF652 and miR-140-3p.This interaction was further validated in PC3 and DU145 cells by interfering circZNF652 as well as in blood sam-ples from PCa and BPH patients.StarBase predicted the binding sites between miR-140-3p and HMGB1,and the HMGB1 expression levels were tested after miR-140-3p overexpression in PC3 and DU145 cells.Western blot was used to detect the HMGB1 expression levels in PCa and BPH patient,PCa cell lines(DU145,PC3)as well as RWPE-1 cells.The dual-luciferase reporter gene assay,RNA immunoprecipitation(RIP)and RNA pull-down were conducted to verified the interaction between miR-140-3p and HMGB1,and the HMGB1expression in PC3 and DU145 cells were detected after transfecting different constructs(si-NC,si-cir-cZNF652#1,si-circZNF652#1+inhibitor NC,si-circZNF652#1+miR-140-3p inhibitor)to further confir-ming that miR-140-3p directly regulates HMGB1.Results The gene chip and qRT-PCR results showed that the expression level of circZNF652 in the plasma of PCa patients was higher than that in BPH patients,com-pared with low-grade PCa and BPH patients,the plasma cirZNF652 expression level in the patients with high grade PCa was upregulated(P<0.001,P<0.01).Compared with normal prostate epithelial cells,the expres-sion levels of circZNF652 in 22RV1,LNCaP,DU145 and PC3 cell lines were upregulated(P<0.05,P<0.05,P<0.001,P<0.01);The OS time in the PCa patients with circZNF652 high expression was shorter than that in PCa patients with circZNF652 low expression.Clinicopathological features analysis revealed that the circ-ZNF652 expression was correlated with the PCa stage(T stage and N stage)and Gleason score.Knocking down circZNF652 expression inhibited the proliferation,invasion and migration in DU145 and PC3 cells.The online database TargetScan predicted that there was the complementary sequences between circZNF652 and miR-140-3p,and dual-luciferase reporter gene and RNA pull-down assays confirmed thatcircZNF652 acted as a sponge for miR-140-3p in PCa cells,interfering circZNF652 experiment in PC3 and DU145 cells also proved this point.The expression of miR-140-3p in the plasma of PCa patients was significantly decreased compared with BPH patients(P<0.05),and the expression level of miR-140-3p was negatively correlate with the ex-pression level of circZNF652(r=-0.888,P<0.001).The database StarBase analysis results showed that there are complementary sequences of HMGB1 mRNA 3'-UTR and miR-140-3p,and the downstream target gene of miR-140-3p was HMGB1.The dual-luciferase reporter gene,RIP and pull-down assays confirmed the direct binding between miR-140-3p and HMGB1.The Western blot experiment found overexpression of miR-140-3p resulted in the HMGB1 expression decrease in DU145 and PC3 cells.Silencing circZNF652 expression resulted in the decrease of HMGB1 protein levels in the DU145 and PC3 cells,while inhibiting miR-140-3p could reverse this decrease.Conclusion Overexpression of circZNF652 could promote the progression and me-tastasis of PCa by regulating the miR-140-3p/HMGB1 axis.
