A novel TNS4 mutant in prostate cancer cells and its mechanism
10.3781/j.issn.1000-7431.2018.11.625
- Author:
Xiubin CHEN
1
Author Information
1. Jiangsu Key Laboratory for Molecular and Medical Biotechnology, School of Life Sciences, Nanjing Normal University
- Publication Type:Journal Article
- Keywords:
Gene expression;
Point mutation;
Prostatic neoplasms;
Signal transducers and activators of transcription;
Tensin family member 4
- From:
Tumor
2018;38(1):1-9
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the expression and mutation status of tensin family member 4 (TNS4) gene in various prostate cancer cell lines, and to explore its potential molecular mechanism. Methods: The expression of TNS4 gene in several prostate cancer cell lines was detected by RT-PCR. The full-length cDNA of TNS4 gene was amplified for the restriction endonuclease hydrolysis and DNA sequencing analysis. Meantime, the recombinant plasmids carrying wild- or mutant-type TNS4 gene were constructed and transfected into prostate cancer PC-3 cells, then the effects of TNS4 over-expression on the mRNA and protein expressions as well as the localization of signal transducer and activator of transcription 1 (STAT1) were detected by RT-PCR, Western blotting and immunofluorescence microscopy, respectively. Results: As compared with benign prostatic hyperplasia BPH1 cells, TNS4 gene was lowly expressed in prostate cancer cell lines DU145, LNCaP and 22Rv1 (all P < 0.01), and nearly undetectable in prostate cancer PC-3 cells. The analysis of full-length TNS4 cDNA sequencing revealed that two sites of single point mutation were identified at T427C and G1455A of TNS4 gene derived from DU145 and LNCaP cells, and the former caused the corresponding amino acid transition (S143P). The recombinant plasmids pcDNA3.1 (+) -TNS4-Mut (TNS4-Mut) and pcDNA3.1 (+) -TNS4-WT (TNS4-WT) were successfully established. After transfection with TNS4-Mut or TNS4-WT, the expression levels of TNS4 mRNA and protein and STAT1 protein in PC-3 cells were significantly increased (all P < 0.05), but the expression level of STAT1 mRNA was not changed (P > 0.05). Additionally, the over-expression of mutant TNS4 significantly promoted nuclear translocation of STAT1 protein (P < 0.01). Conclusion: TNS4 gene is lowly expressed in prostate cancer cell lines, and there is a novel TNS4 mutant (S143P). TNS4 over-expression can up-regulate the expression of STAT1 protein, furthermore the mutant TNS4 can alter the location of STAT1 protein.
