Quantificação dos transcritos do gene fator de alongamento eucariótico 1a (eEF1a) em amostras de pacientes e em linhagens celulares prostáticas

Abstract

Prostate cancer (PCa) is currently the second non-cutaneous malignant tumor in men, accounting for about 29% of the new diagnoses. Different molecular markers have been described for PCa, although still lacking specificity. The eukaryotic elongation factor 1 (eEF1A) gene is a promising one, since it is involved with the regulation of translation elongation. However, the role of this gene in PCa has not yet been fully elucidated. For this reason, we evaluated the transcriptional levels of eEF1A in samples from patients with PCa benign prostatic hyperplasia (BPH) and in prostatic tumor cell lines by qPCR. mRNA levels were correlated with the diagnosis of patients and with cell subtypes. For this, total RNAs were extracted from blood samples, prostatic tissues and from PC3 (castration resistant), LNCaP (hormone-dependent) and RWPE-1 (non-neoplastic) cell lines. The comparative Cq method was used and data were normalized with the reference gene B-2-microglobulin (B2M). The relative expression of eEF1A mRNA in tissue samples was significantly higher in PCa (1.43 fold) compared to BPH (p <0.05). The cut-off of 0.71 was established for the transcripts based on the ROC curve, with sensitivity of 73.68% and specificity of 60.0%. After categorization, patients with relative mRNA levels higher than 0.71 presented 4.2-fold higher chance to develop PCa (CI: 1.19-14.81; p <0.05). The accuracy was determined by the AUC and was defined as 0.70 (p = 0.02). No correlation was found between the transcriptional levels of eEF1A in tissue and the PSA, Gleason and tumor invasion data. The results for the blood samples showed no difference between groups and did not correlate with clinicopathological data. No statistically significant difference was found among the tumor lines. Our data suggest that eEF1A is an essential marker for malignant transformation of prostatic cells and is not involved in its progression. However, functional studies are needed to validate our hypothesis.