Experimental study on effect of circ-DCAF7 on proliferation and migration of prostate cancer cells by regulating miRNA-18a-5p
10.3760/cma.j.cn115355-20250323-00150
- VernacularTitle:circ-DCAF7通过调控miRNA-18a-5p影响前列腺癌细胞增殖和迁移的实验研究
- Author:
Jia LIU
1
;
Yunfei ZHAO
;
Xiaoying WANG
;
Fang XIE
;
Geng HUANG
;
Hong WANG
Author Information
1. 黄石市中心医院 湖北理工学院附属医院超声影像科，黄石 435000
- Publication Type:Journal Article
- Keywords:
Prostatic neoplasms;
RNA, circular;
MicroRNAs;
Cell proliferation;
Cell movement;
circ-DCAF7;
miRNA-18a-5p
- From:
Cancer Research and Clinic
2025;37(9):641-647
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the relationship between circular RNA (circRNA) circ-DCAF7 and the survival of prostate cancer patients, as well as the effects and potential mechanisms of circ-DCAF7 on proliferation and migration of prostate cancer cells in vitro.Methods:The expression and survival data of circ-DCAF7 gene were obtained from 151 prostate cancer patients using the PROGgeneV2 online platform. Patients were divided into circ-DCAF7 high and low expression groups based on the median expression level of circ-DCAF7 gene, and the difference in overall survival between the two groups was compared. Real-time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the expression of circ-DCAF7 at transcription level in human prostate cancer cell lines 22Rv1, VCaP, PC-3, DU-145, LNCaP, and human normal prostate epithelial cell line RWPE-1. The control plasmid (carrying irrelevant control sequences) and the plasmid carrying circ-DCAF7 sequences were transfected into VCaP cells using liposome reagents, respectively, and referred to as the control group and circ-DCAF7 group. The proliferation and migration abilities of two groups of VCaP cells were detected by plate clone formation assay and cell scratch assay, respectively; using CircInteractome online software, specific binding sites were predicted between circ-DCAF7 and miRNA-18a-5p (miR-18a-5p) sequences, and validated using dual luciferase reporter gene assay; qPCR was used to detect the expression of miR-18a-5p at transcription level in VCaP cells of control group and circ-DCAF7 group; Western blotting was used to detect the expression levels of AKT signaling pathway factors p-AKT, p-mTOR, HIF-1α, and c-myc proteins in two groups of VCaP cells.Results:Analysis using the PROGgeneV2 online tool showed that the overall survival of prostate cancer patients in the circ-DCAF7 high expression group was better than that in the circ-DCAF7 low expression group, and the difference was statistically significant ( P < 0.001). Compared with the normal prostate epithelial cell line RWPE-1, the relative expression of circ-DCAF7 at transcription level in all prostate cancer cell lines was lower, and the differences were statistically significant (all P < 0.05). Among them, the expression level of circ-DCAF7 in VCaP cells was the lowest ( P < 0.001). The relative expression of circ-DCAF7 at transcription level in VCaP cells in the circ-DCAF7 group was higher than that in the control group (9.88±1.58 vs. 1.04±0.39), and the difference was statistically significant ( t = 5.42, P = 0.002). The number of VCaP cell clone formation in the circ-DCAF7 group was less than that in the control group (35±12 vs. 116±11), and the difference was statistically significant ( t = 4.96, P = 0.003). The 25-hour scratch healing rate of VCaP cells in the circ-DCAF7 group was lower than that in the control group [(16±3)% vs. (52±6)%], and the difference was statistically significant ( t = 5.37, P = 0.002). The dual luciferase reporter gene assay showed that compared with VCaP cells co-transfected with miR-18a-5p unrelated control and carrying wild-type circ-DCAF7 sequence plasmid, the relative luciferase activity of VCaP cells co-transfected with miR-18a-5p and carrying wild-type circ-DCAF7 sequence plasmid was lower, and the difference was statistically significant ( t = 7.31, P < 0.001). The relative expression of miR-18a-5p at transcription level in VCaP cells in the circ-DCAF7 group was lower than that in the control group (0.99±0.15 vs. 7.55±1.12), and the difference was statistically significant ( t = 5.83, P = 0.001). Western blotting analysis showed that the expression levels of p-AKT, p-mTOR, HIF-1α, and c-myc proteins of the AKT signaling pathway in VCAP cells of the circ-DCAF7 group were lower than those in the control group. Conclusions:The low expression level of circRNA circ-DCAF7 may be associated with poor prognosis of prostate cancer patients; upregulation of circ-DCAF7 level in prostate cancer cells may inhibit cell proliferation and migration by regulating the miR-18a-5p/AKT signaling pathway.
