Overexpression of TRIM2 suppresses proliferation, migration and stemness of clear cell renal cell carcinoma cells in vitro

Abstract

Objective To explore the effect of tripartite motif-containing protein 2 (TRIM2) on the proliferation, migration and stemness of clear cell renal clear carcinoma. Methods We performed a survival analysis of the patients with clear cell renal cell carcinoma using TCGA database to assess how TRIM2 expression affected the clinical outcomes of the patients. We also collected the clinical specimens of clear cell renal cell carcinoma tissues and paired adjacent tissues for testing the expression of TRIM2 using immunohistochemistry. From different clear cell renal clear carcinoma cells, we selected 2 cell lines that expressed low levels of TRIM2 based on the results of Western blotting, and tested the effect of TRIM2 overexpression on the cell proliferation, migration and sphere-forming ability using CCK8 assay, wound healing assay and cell sphere formation assay; the effect of TRIM2 overexpression on the stemness of the cells was evaluated using quantitative real-time PCR and Western blotting. Based on the data from COEXPEDIA database, we constructed a gene co-expression network associated with TRIM2 in renal cancer to explore the role of TRIM2 in the tumorigenesis and progression of clear cell renal cell carcinoma. Results Analysis of the TCGA database showed that in patients with clear cell renal cell carcinoma, a high expression of TRIM2 was associated with a higher survival possibility (P < 0.001). The results of immunohistochemistry showed that TRIM2 was lowly expressed in renal clear cell carcinoma tissues as compared with the adjacent tissues. In clear cell renal cell carcinoma cells with low TRIM2 expression, overexpression of TRIM2 significantly suppressed the cell proliferation (P < 0.01), migration (P < 0.01) and cell sphere formation (P < 0.01) and obviously lowered the expressions of the genes (C-MYC, NANOG, and OCT4) associated with the cell stemness (P < 0.01). Conclusion Overexpression of TRIM2 in clear cell renal cell carcinoma cells can inhibit the cell proliferation, migration and sphere formation and down-regulate the expressions of the stemness genes. The genes (NR3C2, ESRRG, COL4A3 and CTDSPL) that are closely related with TRIM2 may all participate in the occurrence and progression of clear cell renal cell carcinoma.

Keywords

• clear cell renal cell carcinoma
• tripartite motif-containing protein 2
• proliferation
• migration
• cell sphere formation
• stemness