Effect of PRND downregulation on proliferation, migration, and invasion of renal cancer cells
10.3760/cma.j.cn112330-20231212-00184
- VernacularTitle:下调PRND对肾癌细胞增殖、迁移、侵袭的影响
- Author:
Yongkang MA
1
;
Jiangshan PENG
;
Jiuwen ZHANG
;
Huaqi YIN
;
Tiejun YANG
Author Information
1. 郑州大学附属肿瘤医院泌尿外科，郑州 450000
- Publication Type:Journal Article
- Keywords:
Carcinoma, Renal cell;
Prion-like protein polypeptide gene;
Cell proliferation;
Cell migration;
Cell invasion
- From:
Chinese Journal of Urology
2025;46(4):287-294
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of PRND downregulation on the proliferation, migration, and invasion capabilities of human renal carcinoma cells.Methods:Clinical and transcriptomic data from renal carcinoma patients were analyzed using the TCGA database, with bioinformatics methods employed for differential gene expression analysis and survival analysis [including overall survival (OS) and disease-free survival (DFS)]. Postoperative pathological specimens from 50 renal carcinoma patients admitted to Affiliated Cancer Hospital of Zhengzhou University between January 2022 and January 2023 were collected for immunohistochemical staining to assess PRND expression in renal carcinoma tissues. Two distinct small interfering RNAs (siRNAs) were used to downregulate PRND expression in renal carcinoma cell lines ACHN and 769P. Quantitative real-time polymerase chain reaction (qPCR) and Western blotting were performed to validate the knockdown efficiency of PRND at the mRNA and protein levels. The proliferation, migration, and invasion capabilities of ACHN and 769P cells were evaluated using the Cell Counting Kit-8 (CCK-8), cell migration assay, and invasion assay, which was compared between the negative control group (NC) and the two PRND knockdown groups (si1 and si2). Western blotting was used to measure the expression levels of MMP-9, E-cadherin, C-myc, Vimentin, β-catenin, and PD-L1 proteins in ACHN and 769P cell lines.Results:TCGA database analysis revealed that PRND expression was significantly higher in renal carcinoma tissues compared with adjacent normal tissues (1.172 vs. 0.383, P<0.01). Survival analysis indicated that high PRND expression was significantly negatively correlated with both OS ( P<0.01) and DFS ( P<0.01). CCK-8 assay results showed no statistically significant differences in cell viability between the experimental and control groups at 6 hours (ACHN-si1: 1.238±0.659, ACHN-si2: 1.437±0.359, ACHN-NC: 3.234±2.165, P>0.05). However, significant differences were observed at 24 hours (ACHN-si1: 5.608±0.716, ACHN-si2: 7.088±0.308, ACHN-NC: 9.764±1.088, P<0.01), 48 hours (ACHN-si1: 40.422±1.419, ACHN-si2: 41.238±2.623, ACHN-NC: 65.823±4.337, P<0.01), and 72 hours (ACHN-si1: 53.667±4.565, ACHN-si2: 54.533±2.572, ACHN-NC: 78.800±0.265, P<0.01). Similar trends were observed in 769P cells (6 hours: P>0.05; 24 hours: P<0.05; 48 and 72 hours: P<0.01). Cell migration assays demonstrated significantly reduced migration in the experimental groups (ACHN-si1: 31±10, ACHN-si2: 62±19, ACHN-NC: 175±45, P<0.01; 769P-si1: 79±16, 769P-si2: 62±14, 769P-NC: 236±77, P<0.05). Invasion assays also showed significant suppression in the experimental groups (ACHN-si1: 13±9, ACHN-si2: 15±8, ACHN-NC: 54±12, P<0.01; 769P-si1: 17±13, 769P-si2: 19±17, 769P-NC: 91±29, P<0.01). Western blotting revealed that C-myc, β-catenin, MMP-9, Vimentin, and PD-L1 protein levels were lower in the experimental groups, while E-cadherin expression was higher compared to the control groups. Conclusions:PRND is significantly overexpressed in renal carcinoma tissues and closely associated with poor patient prognosis. Downregulation of PRND markedly inhibits the proliferation, migration, and invasion of renal carcinoma cells, potentially through modulation of epithelial-mesenchymal transition (EMT)-related proteins and key molecules involved in tumor metastasis.
