Perfil transcricional do gene ARID1A em células de Câncer de Próstata

• Autor(a) principal: Martins, Isabella Castro
• Data de Publicação: 2018
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UFU
• Texto Completo: https://repositorio.ufu.br/handle/123456789/25344
• topic: Epigenética; Epigenetics; qPCR; Marcador molecular; Molecular marker; Remodelagem de cromatina; Remodeling of chromatin; Complexo SWI/SNF; SWI/SNF complex; CNPQ::CIENCIAS BIOLOGICAS::GENETICA

Prostate cancer (CaP) is the second most common cancer among Brazilian men, after non-melanoma skin cancer. The onset and progression of this malignant neoplasm are associated with several molecular events that control the expression of different genes. The ARID1A protein is one of the subunits that confers specificity to the SWI / SNFA (BAF) chromatin remodeling complex, described as a tumor suppressor. However, its role in PCa has not yet been elucidated. In this sense, we analyzed the role of ARID1A gene transcripts in prostatic tumorigenesis by qPCR. We also correlated the expression of ARID1A with the mRNA levels of the ANXA1, AR, PSA, AR-V7 and HER2 genes. Total mRNA was extracted from LNCaP (hormone-dependent), DU145 and PC3 (hormone-independent) and non-tumorigenic RWPE-1 prostatic cell lines. The comparative Cq method was optimized to quantify the transcripts normalized by the B2M gene. The expression of ARID1A was 1.22 and 1.15 fold higher in LNCaP and PC3 lineages when compared to RWPE-1 (P<0.05; P<0.01, respectively). The mRNA expression of ANXA1 was downregulated in the hormone-dependent lineage, with significantly higher transcripts in DU145 and PC3. Transcripts of the AR and PSA genes were detected only in the LNCaP cell line (P<0.001; P<0.0001). The expression of AR-V7 increased in tumor cell lines, with higher transcriptional levels in LNCaP, compared to DU145 (20.13-fold) and PC3 (4.64-fold) (P<0.01; P<0.01). HER2 expression was also upregulated in LNCaP with increased transcripts (8.43-fold, 9.34-fold and 4.43-fold) when compared to RWPE-1, DU145 and PC3, respectively (P<0.0001; P<0.0001; P<0.001). The ARID1A transcripts did not correlate with the others. A negative correlation was verified between ANXA1 and the AR, PSA, AR-V7 and HER2 targets. The latter four presented a positive correlation with each other (R>0.94). The results suggest that ARID1A is involved in the onset or initial development of PCa. The loss of ANXA1 in LNCaP may be related to the tumor's genesis and the subsequent elevation of mRNA levels in DU145 and PC3 may be associated to the resistance to hormone therapy. Additional experiments are needed to evaluate the behavior of the proteins transcribed by these genes for validation of biomarkers to advanced PCa.