The involvement of miR-100 in bladder urothelial carcinogenesis changing the expression levels of mRNA and proteins of genes related to cell proliferation, survival, apoptosis and chromosomal stability

Cancer Cell International

Katia RM Leite¹  Miguel Srougi¹  Iran A Silva¹  Nelson Dip¹  Caio Moura¹  Cristina Massoco²  Camila Berfort Piantino¹  Nayara Viana¹  Sabrina T Reis¹  Denis R Morais² 

[1] Laboratory of Medical Research, Department of Urology – LIM55, University of Sao Paulo Medical School, Sao Paulo, Brazil;Department of Pathology, University of Sao Paulo Veterinary Medicine and Zootechnics School, Sao Paulo, Brazil

关键词: Western blotting;    PCR;    Protein expression;    Gene expression;    miR-100;    MicroRNA;    Bladder cancer;   

Others  :  1121630
DOI  :  10.1186/s12935-014-0119-3

 received in 2014-02-26, accepted in 2014-10-30,  发布年份 2014

【 摘 要 】

Introduction

MicroRNAs (miRNA) are small non-coding RNAs that play an important role in the control of gene expression by inhibiting protein translation or promoting messenger RNA degradation. Today, miRNAs have been shown to be involved in various physiological and pathological cellular processes, including cancer, where they can act as oncogenes or tumor suppressor genes. Recently, lowered expression of miR-100, resulting in upregulation of FGFR3, has been correlated with low-grade, non-invasive bladder urothelial cancer, as an alternative oncogenesis pathway to the typical FGFR3 gene mutation. Our aim is to analyze the role of miR-100 in bladder cancer cell lines in controlling the expression of some of its possible target genes, including FGFR3 and its relationship with proliferation, apoptosis and DNA ploidy.

Methods

The bladder cancer cell lines RT4 and T24 were transfected with pre-miR 100, anti-miR 100 and their respective controls using a lipid-based formulation. After transfection mRNA and protein levels of its supposed target genes THAP2, BAZ2A, mTOR, SMARCA5 and FGFR3 were analyzed by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. Cell proliferation, apoptosis and DNA ploidy were analyzed by flow cytometry. For statistical analysis, a t-test was applied, p < 0.05 was considered significant.

Results

After miR-100 transfection, there was a significant reduction in the mRNA of mTOR (p = 0.006), SMARCA5 (p = 0.007) and BAZ2A (p = 0.029) in RT4, mTOR (p = 0.023) and SMARCA5 (p = 0.015) in T24. There was a reduction in the expression of all proteins, variable from 22.5% to 57.1% in both cell lines. In T24 miR-100 promoted an increase in cell proliferation and anti-miR 100 promoted apoptosis characterizing miR-100 as an oncomiR in this cell line representative of a high-grade urothelial carcinoma.

Conclusion

miR-100 transfection reduces expression of BAZ2A, mTOR and SMARCA5 mRNA and protein in BC cell lines. miR-100 would be classified as an oncomiR in T24 cells representative of high grade urothelial carcinoma promoting increase in cell proliferation and reduction in apoptosis. The knowledge of miRNA role in tumors will allow their use as tumor markers and targets for new therapies.

【 授权许可】

   
2014 Morais et al.; licensee BioMed Central Ltd.

【 图 表 】

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