Mauricio Rodríguez-Dorantes

miRNA biogenesis: biological impact in the development of cancer.

microRNAs (miRNAs) are non coding RNAs with different biological functions and pathological implications. Given their role as post-transcriptional gene expression regulators, they are involved in several important physiological processes like development, cell differentiation and cell signaling. miRNAs act as modulators of gene expression programs in different diseases, particularly in cancer, where they act through the repression of genes which are critical for carcinogenesis. The expression level of mature miRNAs is the result of a fine mechanism of biogenesis, carried out by different enzymatic complexes that exert their function at transcriptional and post-transcriptional levels. In this review, we will focus our discussion on the alterations in the miRNA biogenesis machinery, and its impact on the establishment and development of cancer programs.

MicroRNAs transported by exosomes in body fluids as mediators of intercellular communication in cancer

Cancer-cell communication is an important and complex process, achieved through a diversity of mechanisms that allows tumor cells to mold and influence their environment. In recent years, evidence has accumulated indicating that cells communicate via the release and delivery of microRNAs (miRNAs) packed into tumor-released (TR) exosomes. Understanding the role and mode of action of miRNAs from TR exosomes is of paramount importance in the field of cancer biomarker discovery and for the development of new biomedical applications for cancer therapeutics. In this review, we focus on miRNAs secreted via TR exosomes, which by acting in a paracrine or endocrine manner, facilitate a diversity of signaling mechanisms between cancer cells. We address their contribution as signaling molecules, to the establishment, maintenance, and enhancement of the tumor microenvironment and the metastatic niche in cancer. Finally, we address the potential role of these molecules as biomarkers in cancer diagnosis and prognosis and their impact as a biomedical tool in cancer therapeutics.

Differential expression of the estrogen-regulated proto-oncogenes c-fos, c-jun, and bcl-2 and of the tumor-suppressor p53 gene in the male mouse chronically infected with Taenia crassiceps cysticerci.

Abstract Chronic infection with Taenia crassiceps cysticerci produces a 200-fold increase in serum estradiol levels in male mice. The aim of this study was to investigate the expression pattern of c-fos and c-jun, two estradiol-regulated genes, as well as that of p53 and bcl2 in the testes, spleen, and thymus of male mice infected with T. crassiceps cysticerci. In parasitized animals the c-fos mRNA content was significantly increased in all tissues studied, whereas the c-jun mRNA content was increased only in the thymus. The p53 mRNA content was markedly reduced in all tissues of the parasitized animals analyzed, whereas bcl-2 gene expression was abolished in the thymus. On the other hand, thymic cell analysis performed by flow cytometry showed a diminution in the content of CD3+, CD4+, and CD8+ subpopulations in the parasitized mice. Our results suggest that the increase in estradiol levels of the host should change the expression pattern of several genes that participate in apoptosis regulation in the thymus of male mice during chronic infection with T. crassiceps cysticerci.

Modified expression of steroid 5α-reductase as well as aromatase, but not cholesterol side-chain cleavage enzyme, in the reproductive system of male mice during (Taenia crassiceps) cysticercosis

Abstract Infection with Taenia crassiceps cysticerci in male mice produces an increase in serum estradiol levels, whereas serum testosterone is abolished. Concomitantly, complete atrophy of the reproductive tract of infected male mice is observed. The present study was undertaken to determine the expression pattern of three key steroidogenic enzymes in the reproductive tissues of normal and infected male mice. In infected mice, serum estradiol levels were increased 97 times as compared with control mice of the same age. Testosterone and dihydrotestosterone levels were completely inhibited. The expression of 5α-reductase in the reproductive tract was markedly reduced, whereas aromatase mRNA levels were highly elevated in the testes of parasitized mice. No change in the mRNA content for cholesterol side-chain cleavage enzyme was evident. The overall results suggest that the change in the normal production of sex steroids in infected male mice is produced concomitantly by the inhibition of expression of the 5α-reductase enzyme and the activation of aromatase gene expression. This induces a preferential metabolism from testosterone to estradiol instead of the normal metabolism from testosterone to dihydrotestosterone.

Tissue damage in the male murine reproductive system during experimental Taenia crassiceps cysticercosis.

Chronic infection with Taenia crassiceps cysticerci in male mice increases the level of estradiol in serum, whereas it reduces that of testosterone. In addition, male mice lose their typical male reproductive behavior. The effects of cysticerci infection on the histomorphology of male reproductive tissues are unknown. The present study was undertaken to determine the histological changes in testes, seminal vesicles, and prostate of male mice infected with T. crassiceps cysticerci. At 16 wk of infection, all tissues exhibited high inflammatory infiltrate. Tissue lesions included marked dilation and peripheral fibrosis. In the testes, a diminution of spermiogenesis was observed. The overall results indicated that the histological changes in chronically parasitized male mice occurred with changes in hormone levels, simultaneously with the high inflammatory immune response.

Urinary microRNA-based signature improves accuracy of detection of clinically relevant prostate cancer within the prostate-specific antigen grey zone

At present, prostate-specific antigen (PSA) is used as a clinical biomarker for prostate cancer (PCa) diagnosis; however, a large number of patients with benign prostate hyperplasia (BPH) with PSA levels in the ʻgray areaʼ (4–10 ng/ml) are currently subjected to unnecessary biopsy due to overdiagnosis. Certain microRNAs (miRs) have been proven to be useful biomarkers, several of which are detectable in bodily fluids. The present study identified and validated a urinary miR-based signature to enhance the specificity of PCa diagnosis and to reduce the number of patients with benign conditions undergoing biopsy. Seventy-three urine samples from Mexican patients with diagnosis of PCa with a Gleason score ≥7 and 70 patients diagnosed with BPH were collected after digital rectal examination (DRE) of the prostate. miR expression profiles were determined using TaqMan Low Density Array experiments, and normalized Ct values for the miRs were compared between PCa and BPH groups. Receiver operating characteristic (ROC) curve analysis was performed to evaluate whether miR detection in urine is suitable for distinguishing patients with PCa from those with BPH. The identified miR-100/200b signature was significantly correlated with PCa. Using a multivariable logistic regression approach, a base model including the clinical variables age, prostate-specific antigen (PSA), the percentage of free PSA and DRE was generated, and a second base model additionally contained the miR-100/200b signature. ROC analysis demonstrated that the combined model significantly outperformed the capacity of PSA (P<0.001) and the base model (P=0.01) to discriminate between PCa and BPH patients. In terms of evaluation of the sub-group of patients in the gray zone of PSA levels, the performance of the combined model for predicting PCa cases was significantly superior to PSA level determination (P<0.001) and the base model (P=0.009). In addition, decision curve analysis demonstrated that the use of the combined model increased the clinical benefit for patients and produced a substantial reduction in unnecessary biopsies across a range of reasonable threshold probabilities (10–50%). Detection of the urinary miR signature identified in the present study as part of clinical diagnostic procedures will enhance the accuracy of PCa diagnosis and provide a clinical benefit for patients with BPH by sparing them from undergoing invasive biopsy. To the best of our knowledge, the present study was the first to describe the profiling of urinary miR100 and miR-200b levels for the clinical diagnosis of PCa.

Taenia crassiceps infection disrupts estrous cycle and reproductive behavior in BALB/c female mice

Previously, it has been shown that parasitic infections are able to alter the normal mammal physiology, at several extents. Thus, we investigated the effects on estrous cycle and sexual behavior induced by intraperitoneal infection with Taenia crassiceps in female host mice. Along the weeks of infection, parasites were collected from the peritoneal cavity of female mice, showing the maximum parasite load at 16 weeks. No parasites were found outside peritoneal cavity. Vaginal estrous cycle was monitored daily for 4, 8, 12 and 16 weeks of infection, and results compared against age-matched female mice. Female sexual behavior (FSB) tests were performed, one test per week. Immediately after the last behavioral test, blood was collected by cardiac puncture for steroid determinations. First of all, there was a strong tissular damage in the female reproductive tract in all infected females. The phases of the estrous cycle were interrupted at 12 and 16 weeks, with increased leukocytes and the presence of a few cornified epithelial cells and nucleated epithelial cells. The FSB decreased starting 6 weeks post infection. On the 16th week, all infected female mice ceased to exhibit sexual responses, and estradiol levels showed a significant decrease. Control mice continued showing FSB and the different phases of the estrous cycle throughout the observation period. Our results strength the notion that parasites may be considered as an evolutionary force in the reproductive ability of mammals.

Synergistic effects of ICI 182,780 on the cytotoxicity of cisplatin in cervical carcinoma cell lines

PurposeWe investigated the ability of the novel pure antiestrogen ICI 182,780 to modulate the cytotoxic effects of cisplatin in several cervical cancer cell lines.MethodsThe effect of cisplatin alone and cisplatin combined with ICI 182,780 on cellular death was studied using an assay based on a tetrazolium dye (sodium 3′-[1-(phenylamino-carbonyl)-3,4-tetrazolium], XTT). Before and after treatment with ICI 182,780, expression of the estrogen and progesterone receptor genes were assessed by a reverse transcriptase polymerase chain reaction (RT-PCR). Cell-cycle modifications after combined treatment with cisplatin and ICI 182,780 were studied by flow cytometry.ResultsAnalysis of the data by the isobologram method showed that the combination of ICI 182,780 and cisplatin produced a synergistic antiproliferative effect in cervical cancer cells. The effect of ICI 182,780 on the cytotoxicity of cisplatin could be mediated, at least partially, by inhibition of estrogen and progesterone gene expression and by arresting the cell cycle at the G2/M phase.ConclusionsOur results suggest that ICI 182,780 can improve the efficacy of cisplatin in cancer cells and that this antihormonal drug therapy may be a useful candidate for further evaluation in combination with antineoplastic drugs, particularly cisplatin, in the treatment of cancer.

SFRP1 repression in prostate cancer is triggered by two different epigenetic mechanisms

Worldwide, prostate cancer (PCa) is the second cause of death from malignant tumors among men. Establishment of aberrant epigenetic modifications, such as histone post-translational modifications (PTMs) and DNA methylation (DNAme) produce alterations of gene expression that are common in PCa. Genes of the SFRP family are tumor suppressor genes that are frequently silenced by DNA hypermethylation in many cancer types. The SFRP family is composed of 5 members (SFRP1-5) that modulate the WNT pathway, which is aberrantly activated in PCa. The expression of SFRP genes in PCa and their regulation by DNAme has been controversial. Our objective was to determine the gene expression pattern of the SFRP family in prostatic cell lines and fresh frozen tissues from normal prostates (NP), benign prostatic hyperplasia (BPH) and prostate cancer (PCa), by qRT-PCR, and their DNAme status by MSP and bisulfite sequencing. In prostatic cancer cell lines, the 5 SFRPs showed significantly decreased expression levels compared to a control normal prostatic cell line (p<0.0001). In agreement, SFRP1 and SFRP5 genes showed decreased expression levels in CaP fresh frozen tissues compared to NP (p<0.01), while a similar trend was observed for SFRP2. Conversely, increased levels of SFRP4 expression were found in PCa compared to BPH (p<0.01). Moreover, SFRP2, SFRP3, and SFRP5 showed DNA hypermethylation in PCa cell lines. Interestingly, we observed DNA hypermethylation at the promoter of SFRP1 in the PC3 cell line, but not in LNCaP. However, in the LNCaP cell line we found an aberrant gain of the repressive histone posttranslational modification Histone H3 lysine 27 trimethylation (H3K27me3). In conclusion, decreased expression by DNA hypermethylation of SFRP5 is a common feature of PCa, while decreased expression of SFRP1 can be due to DNA hypermethylation, but sometimes an aberrant gain of the histone mark H3K27me3 is observed instead.

Allopregnanolone promotes proliferation and differential gene expression in human glioblastoma cells

&NA; Allopregnanolone (3&agr;‐THP) is one of the main reduced progesterone (P4) metabolites that is recognized as a neuroprotective and myelinating agent. 3&agr;‐THP also induces proliferation of different neural cells. It has been shown that P4 favors the progression of glioblastomas (GBM), the most common and aggressive primary brain tumors. However, the role of 3&agr;‐THP in the growth of GBMs is unknown. Here, we studied the effects of 3&agr;‐THP on the number of cells, proliferation and gene expression in U87 cell line derived from a human GBM. 3&agr;‐THP (10, 100 nM and 1 &mgr;M) increased the number of U87 cells, and at 10 nM exerted a similar increase in both the number of total and proliferative U87 cells as compared with P4 (10 nM). Interestingly, finasteride (F; 100 nM), an inhibitor of 5&agr;‐reductase (5&agr;R), an enzyme necessary to metabolize P4 and produce 3&agr;‐THP, blocked the increase in the number of U87 cells induced by P4. By using RT‐qPCR, we determined that U87 cells express 5&agr;‐R isoenzymes 1 and 2 (5&agr;R1 and 5&agr;R2), being 5&agr;R1 the predominant one in these cells. 3&agr;‐THP (10 nM) increased the expression of TGF&bgr;1, EGFR, VEGF and cyclin D1 genes. P4 increased TGF&bgr;1 and EGFR expression, and this effect was blocked by F. These data provide evidence that P4, through its metabolite 3&agr;‐THP, can promote in part cell proliferation of human GBM cells by changing the expression of genes involved in tumor progression. Highlights3&agr;‐THP and progesterone (P4) promote proliferation of U87 human glioblastoma cells.U87 cells express higher levels of 5&agr;R1 than those of 5&agr;R2.P4 increases TGF&bgr;1 and EGFR expression, and finasteride blocks this effect.3&agr;‐THP promotes the expression of TGF&bgr;1, EGFR, VEGF and cyclin D1 in U87 cells.