Molecular Biology, Biochemistry, Genetics and Molecular Biology
34
Scopus Publications
Scopus Publications
MiR-10a as a Potential Biomarker and Therapeutic Target in Localized and Metastatic Prostate Cancer Tiago José Borelli Bovo, Juliana Alves de Camargo, Ruan Pimenta, Vanessa Ribeiro Guimarães, Patrícia Candido, Katia Ramos Moreira Leite, Carlo Camargo Passerotti, William Carlos Nahas, Sabrina T. Reis Current Issues in Molecular Biology, 2025 Introduction: Prostate cancer (PC) accounts for around 10% of all cancers worldwide and is the fourth most common neoplasm. Localized PC has high cure rates when diagnosed early, but 35% of patients progress to the metastatic form. The search for new molecular markers, such as microRNAs, is fundamental to improving diagnosis and treatment. The role of miR-10a is controversial between tumor tissues, opening a niche for studies on their role in PC. Objectives: To evaluate the role of miR-10a in metastatic PC cell lines, focusing on the mechanisms of proliferation, migration, and invasion, and to analyze the expression in surgical specimens of localized PC. Methods: Three commercial metastatic PC cell lines were used: LNCaP, DU145 and PC-3. Expression of mimic miR-10a was induced by cell transfection, followed by extraction of miRNA and total RNA. The synthesis of complementary DNA (cDNA) and analysis by real-time PCR enabled the expression of miR-10a and the VEGF, MYC, and HAS3 genes to be assessed. Matrigel, colony formation, invasion, and migration assays were evaluated for the transfected cells. The surgical specimens were used to evaluate the miR-10a expression. Results: Transfected cells with mimic significantly increased the expression of miR-10a in the LNCaP (p = 0.0179), PC-3 (p ≤ 0.001), and DU145 (p ≤ 0.001) cell lines. Transfected cells reduced cell invasion in the PC-3 (p = 0.001) and DU-145 (p = 0.0004) cell lines and decreased cell migration and proliferation. In surgical specimens, miR-10a expression was higher in PC compared to Benign Prostatic Hyperplasia (p = 0.0010). Conclusions: Increased expression of miR-10a affects cell migration, invasion, and proliferation, showing potential as a therapeutic target in treating PC.
Enhancing RECK Expression Through miR-21 Inhibition: A Promising Strategy for Bladder Carcinoma Control Paulo Rodolfo Moraes dos Santos, Paulo Ricardo da Silva Gomes, Poliana Romão, Feres Camargo Maluf, Vanessa Ribeiro Guimarães, Patrícia Candido, Guilherme Lopes Gonçalves, Juliana Alves de Camargo, Gabriel Arantes dos Santos, Iran Silva, Katia Ramos Moreira Leite, William Nahas, Sabrina T. Reis, Ruan Pimenta, Nayara Izabel Viana Biochemical Genetics, 2025
MicroRNA-29b attenuates fibrosis in a rat model of Peyronie's disease Patrícia Candido, Ruan Pimenta, Feres Camargo Maluf, Caroline Chiovatto, Poliana Romão, Camila Machado Baldavira, Vitória Ghazarian, Juliana A. Camargo, Vanessa R. Guimarães, Gabriel A. dos Santos, Iran A. Silva, Bruno Nascimento, Jorge Hallak, Vera Luiza Capelozzi, Miguel Srougi, William C. Nahas, Nayara I. Viana, Katia R. Leite, Sabrina T. Reis Andrology, 2025 BackgroundPeyronie's disease is characterized by the formation of fibrotic plaques in the penile tunica albuginea. Effective treatments are limited, warranting the investigation of new promising therapies, such as the application of microRNAs that regulate fibrosis‐related genes.ObjectiveWe aimed to investigate the therapeutic potential of mimicking microRNA‐29b in a fibrin‐induced rat model of Peyronie's disease.Material/methodsThe study was designed in two phases. To establish an optimal Peyronie's disease model, rats received either human fibrin and thrombin or saline solutions into the tunica albuginea on days 0 and 5. The animal model validation was done through expression and histopathological analyses, the latest by an experienced uropathologist. After validation, we performed microRNA‐29b treatment on days 14, 21, and 28 of the study. This phase had control (normal saline) and scramble (microRNA scramble) groups. The mid‐penile shaft was removed on day 30 for histological examination and molecular analyses in both study stages.ResultsThe control group displayed typical tunica albuginea histologic architecture in the animal model validation. In Peyronie's disease group, the Hematoxylin and eosin and Masson Trichrome staining methods demonstrated an interstitial inflammatory process with concomitant dense fibrotic plaques as well as disarrangement of collagen fibers. Additionally, we found out that reduced microRNA‐29b (p = 0.05) was associated with significantly increased COL1A1 and transforming growth factor β1 genes and proteins (p > 0.05) in the Peyronie's disease group. After treatment with mimic microRNA‐29b stimulation, the Hematoxylin & eosin and Masson Trichrome staining revealed a discrete and less dense fibrotic plaque. This result was associated with significantly decreasing expression of COL1A1, COL3A1, and transforming growth factor β1 genes and proteins (p < 0.05).DiscussionThe fibrin‐induced animal model showed significant histopathological and molecular changes compared to the Control group, suggesting that our model was appropriate. Previous findings have shown that increased expression of microRNA‐29b was associated with decreased pathological fibrosis. In the present study, treatment with microRNA‐29b decreased the gene and protein expression of collagens and transforming growth factor β1. This study reveals the therapeutic potential for Peyronie's disease involving molecular targets.ConclusionMicroRNA‐29b application on the rat's tunica albuginea attenuated fibrosis, arising as a novel potential strategy for Peyronie's disease management.
CRISPR/Cas9–mediated MMP-9 silencing inhibits bladder cancer T24 cell invasion and migration in vitro Fabio Pescarmona Gallucci, Juliana Alves de Camargo, Nayara Izabel Viana, Ruan Cesar Aparecido Pimenta, Vanessa Ribeiro Guimarães, Patrícia Candido, Katia Ramos Moreira Leite, William Carlos Nahas, Sabrina Thalita dos Reis Clinics, 2025 • CRISPR/Cas9 effectively silenced MMP-9 in bladder cancer cells. • MMP-9 silencing reduced T24 cell proliferation and colony formation. • Edited cells showed increased apoptosis after MMP-9 knockdown. • Migration and invasion capacity were strongly inhibited by MMP-9 silencing. • MMP-9 is a potential therapeutic target in bladder cancer progression. Bladder Cancer (BCa) ranks as the tenth most common cancer worldwide, with high morbidity and mortality. Matrix Metalloproteinases (MMPs), especially MMP-9, are associated with tumor progression and metastasis. This study aimed to evaluate the effects of MMP-9 silencing using CRISPR-Cas9 in T24-luc bladder carcinoma cells. Guide RNAs (sgRNA1 and sgRNA2) targeting MMP-9 were cloned into pX-330 plasmids and transfected into T24-luc cells. Gene and protein expression were analyzed via RT-qPCR and Western blotting. Functional assays included flow cytometry for proliferation and apoptosis, colony formation, wound healing, and Matrigel™ invasion assays. sgRNA2 significantly reduced MMP-9 gene expression, while both sgRNAs reduced protein expression. Edited cells showed decreased proliferation and colony formation, increased apoptosis, and reduced migration and invasion capacity. CRISPR-Cas9-mediated silencing of MMP-9 inhibited cell proliferation, migration, invasion, and increased apoptosis in BCa cells, supporting that MMP-9 has an important effect on the progression of bladder cancer.
Interconnected study of molecular pathways: miR-137 as a central element at the intersection of lipid metabolism and prostate carcinogenesis Karina Serafim da Silva, Vanessa Ribeiro Guimarães, Feres Camargo Maluf, Gabriel Arantes dos Santos, Juliana Alves de Camargo, Iran Amorim da Silva, Katia Ramos Moreira Leite, Sabrina Thalita dos Reis, Nayara Izabel Viana, Miguel Srougi, Ruan Pimenta Einstein Sao Paulo Brazil, 2025 OBJECTIVE: To evaluate the roles of miR-137 and its target genes in lipid metabolism and prostate tumorigenesis. METHODS: We used a series of bioinformatic approaches to establish the relationship between miR-137 and its target genes. We mapped the metabolic pathways of interest in the Reactome database and identified the central target genes of miR-137 in this pathway using four platforms: Reactome, miRDB, miRmap, and TargetScan. To assess the expression and association with clinical parameters, we obtained information from the UALCAN, OncoDB, and GEPIA2 databases using a dataset of patients with prostate cancer from The Cancer Genome Atlas. For functional enrichment analysis and construction of the protein-protein interaction network, we used the Kyoto Encyclopedia of Genes and Genomes, Gene Ontology, and STRING. RESULTS: Our in silico study of The Cancer Genome Atlas database revealed that miR-137 is underexpressed in tumor tissues, and its reduction is associated with poor prognosis. An intriguing set of eight genes within the PPARα pathway: PPARGC1A, PPARGC1B, NCOA1, NCOA2, NCOA3, MED1, MED27, and ESRRA displayed synergy, positive correlations, and synchronized expression patterns in adipose, hepatic, and prostatic tissues, all linked to the enigmatic processes of metabolic regulation. Among the highlighted genes, ESRRA was overexpressed in the malignant environment, whereas its counterparts remained underexpressed. The plot was thickened with associations between the expression of NCOA1, NCOA3, and MED27, lymph node involvement, and the overexpression of several genes linked to advanced prostate cancer stages. An intriguing pattern emerged, with patients exhibiting reduced disease-free survival overexpressing NCOA2, NCOA3, MED27, and ESRRA. CONCLUSION: This study elucidates the possibility that miR-137 subtly modulates metabolic genes in prostate cancer, suggesting its latent therapeutic potential as a biomarker for disease progression. BACKGROUND: ■ The reduction of miR-137 in tumor tissues is associated with a worse prognosis. BACKGROUND: ■ miR-137 has eight oncogenically relevant target genes acting in the PPARα lipid pathway. BACKGROUND: ■ NCOA1, NCOA3, MED27, and ESRRA are associated with advanced prostate cancer. BACKGROUND: ■ miR-137 exhibits significant clinical potential by repressing the activation of pathways that influence prostate tumorigenesis in hyperstimulated metabolic environments. BACKGROUND: Prostate cancer progression is sustained by the simultaneous activation of pathways involving lipid uptake and de novo synthesis. In this context, miR-137 inhibits adipogenic differentiation and may reduce lipid uptake by tumor cells by modulating the PPAR/ p160/ESRRA axis, considerably attenuating metabolic effects and suppressing prostate tumorigenesis.
Is miR-10a a tumor suppressor that modulates proliferation and invasion in high-grade bladder cancer? THAINá RODRIGUES, PATRíCIA CANDIDO, FERES CAMARGO MALUF, POLIANA ROMãO, CAROLINA MIE MIOSHI, VANESSA RIBEIRO GUIMARãES, JULIANA ALVES DE CAMARGO, KARINA SERAFIM DA SILVA, GABRIEL ARANTES DOS SANTOS, IRAN AMORIM SILVA, KATIA RAMOS MOREIRA LEITE, WILLIAM C. NAHAS, SABRINA T. REIS, RUAN PIMENTA, NAYARA IZABEL VIANA Oncology Research, 2025 Objectives: Bladder Cancer (BC) is one of the most commonly diagnosed malignancies worldwide, with high rates of mortality and morbidity. It can be classified as non-muscle invasive bladder cancer (NMIBC) or muscle-invasive bladder cancer (MIBC), with radical cystectomy being the treatment for MIBC, which significantly reduces quality of life. MicroRNAs (miRs) act as critical genetic regulators, with both oncogenic and tumor-suppressive roles. MiR-10a is described as a tumor suppressor in various neoplasms, but its role in BC is controversial. This study aims to assess the activity of miR-10a in cellular invasion and proliferation in two distinct BC cell lines. Methods: The study used high-grade T24 and low-grade RT4 bladder cell lines. Cells were transfected with miR-10a mimic or a non-targeting control. Transfection efficiency was validated by qPCR. Cell proliferation was cultured for 10–14 days. Cell migration and invasion were evaluated using Matrigel. All assays were conducted in triplicate. Results: The T24 cells transfected with miR-10a presented decreased cellular proliferation and invasion compared to the Scramble (p = 0.0481 and p < 0.0001, respectively). In the RT4 cell line, there was only a significant reduction in cellular proliferation after miR-10a transfection (p = 0.0029). Conclusions: Our findings suggest that miR-10a has a tumoral suppressor role in BC, demonstrating higher efficacy in high-grade cells.
Phospholipase A2 expression in prostate cancer as a biomarker of good prognosis: A comprehensive study in patients with long follow-up Saulo da Cunha Recuero, Nayara I Viana, Sabrina T Reis, Keith T Mendes, Leda L Talib, Wagner F Gattaz, Vanessa R Guimarães, Iran A Silva, Ruan C. P Pimenta, Juliana A Camargo, Willian C Nahas, Miguel Srougi, Katia R. M Leite Urologia Journal, 2024 Background: Phospholipase A2 (PLA2) is a large family of enzymes involved in the inflammatory process that catalyzes the hydrolysis of membrane phospholipids, leading to the production of free fatty acids and lysophospholipids, starting the arachidonic acid cascade. Their expression has been related to the behavior of several cancers. Our objective is to search for PLA2 expression in prostate cancer (PCa) tissue that correlates with prognosis and survival. Methods: Using qRT-PCR, we analyzed the expression levels of PLA2G1B, PLA2G2A, PLA2G2D, PLA2G4A, PLA2G4B, PLA2G4C, PLA2G4D, PLA2G4E, PLA2G4F, PLA2G6, PLA2G7, PLA2G16, PNPLA1, and PNPLA2 in PCa tissue from 108 patients submitted to radical prostatectomy, followed by a mean time of 163 months. Results: All PLA2 was overexpressed in PCa compared to normal tissue. Interestingly, higher expression of some PLA2 was related to favorable prognostic factors: lower levels of PSA (PLA2G2A, PLA2G4D), lower rates of lymph node metastasis (PLA2G16 and PLA2G1B), and organ-confined disease (PLA2G4A). Most importantly, PLAG4B was independently related to longer disease-free survival. Conclusion: This is the first study exploring comprehensively the expression levels of PLA2 in PCa, showing that the higher expression of some PLA2 should be used as biomarkers of good prognosis and longer disease-free survival.
Evaluation of AR, AR-V7, and p160 family as biomarkers for prostate cancer: insights into the clinical significance and disease progression Ruan Pimenta, Feres Camargo Malulf, Poliana Romão, Giovana Vilas Boas Caetano, Karina Serafim da Silva, Vitoria Ghazarian, Gabriel A. dos Santos, Vanessa Guimarães, Iran Amorim Silva, Juliana Alves de Camargo, Saulo Recuero, Bárbara V. Lima Aguiar Melão, Alberto Azoubel Antunes, Miguel Srougi, William Nahas, Katia R. M. Leite, Sabrina T. Reis Journal of Cancer Research and Clinical Oncology, 2024 Purpose To assess the role of the p160 family, AR, and AR-V7 in different initial presentations of prostate cancer and their association with clinical endpoints related to tumor progression. Methods The study sample comprises 155 patients who underwent radical prostatectomy and 11 healthy peripheral zone biopsies as the control group. Gene expression was quantified by qPCR from the tissue specimens. The statistical analysis investigated correlations between gene expression levels, associations with disease presence, and clinicopathological features. Additionally, ROC curves were applied for distinct PCa presentations, and time-to-event analysis was used for clinical endpoints. Results The AR-V7 diagnostic performance for any PCa yielded an AUC of 0.77 (p < 0.05). For locally advanced PCa, the AR-V7 AUC was 0.65 (p < 0.05). Moreover, the metastasis group had a higher expression of SRC-1 than the non-metastatic group (p < 0.05), showing a shorter time to metastasis in the over-expressed group (p = 0.005). Patients with disease recurrence had super-expression of AR levels (p < 0.0005), with a shorter time-to-recurrence in the super-expression group (p < 0.0001). Conclusion Upregulation of SRC-1 indicates a higher risk of progression to metastatic disease in a shorter period, which warrants further research to be applied as a clinical tool. Additionally, AR may be used as a predictor for PCa recurrence. Furthermore, AR-V7 may be helpful as a diagnostic tool for PCa and locally advanced cancer, comparable with other investigated tools.
The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer Juliana A. Camargo, Nayara I. Viana, Ruan Pimenta, Vanessa R. Guimarães, Gabriel A. dos Santos, Patrícia Candido, Vitória Ghazarian, Poliana Romão, Iran A. Silva, Alexander Birbrair, Miguel Srougi, William C. Nahas, Kátia R. Leite, Ericka B. Trarbach, Sabrina T. Reis International Journal of Molecular Sciences, 2023 Prostate cancer (PCa) has a high prevalence and represents an important health problem, with an increased risk of metastasis. With the advance of CRISPR-Cas9 genome editing, new possibilities have been created for investigating PCa. The technique is effective in knockout oncogenes, reducing tumor resistance. MMP9 and miR-21 target genes are associated with PCa progression; therefore, we evaluated the MMP-9 and miR-21 targets in PCa using the CRISPR-Cas9 system. Single guide RNAs (sgRNAs) of MMP9 and miR-21 sequences were inserted into a PX-330 plasmid, and transfected in DU145 and PC-3 PCa cell lines. MMP9 and RECK expression was assessed by qPCR, WB, and IF. The miR-21 targets, integrins, BAX and mTOR, were evaluated by qPCR. Flow cytometry was performed with Annexin5, 7-AAD and Ki67 markers. Invasion assays were performed with Matrigel. The miR-21 CRISPR-Cas9-edited cells upregulated RECK, MARCKS, BTG2, and PDCD4. CDH1, ITGB3 and ITGB1 were increased in MMP9 and miR-21 CRISPR-Cas9-edited cells. Increased BAX and decreased mTOR were observed in MMP9 and miR-21 CRISPR-Cas9-edited cells. Reduced cell proliferation, increased apoptosis and low invasion in MMP9 and miR-21 edited cells was observed, compared to Scramble. CRISPR-Cas9-edited cells of miR-21 and MMP9 attenuate cell proliferation, invasion and stimulate apoptosis, impeding PCa evolution.
Metformin acts in the gut and induces gut-liver crosstalk Natália Tobar, Guilherme Z. Rocha, Andrey Santos, Dioze Guadagnini, Heloísa B. Assalin, Juliana A. Camargo, Any E. S. S. Gonçalves, Flavia R. Pallis, Alexandre G. Oliveira, Silvana A. Rocco, Raphael M. Neto, Irene Layane de Sousa, Marcos R. Alborghetti, Maurício L. Sforça, Patrícia B. Rodrigues, Raissa G. Ludwig, Emerielle C. Vanzela, Sergio Q. Brunetto, Patrícia A. Boer, José A. R. Gontijo, Bruno Geloneze, Carla R. O. Carvalho, Patricia O. Prada, Franco Folli, Rui Curi, Marcelo A. Mori, Marco A. R. Vinolo, Celso D. Ramos, Kleber G. Franchini, Claudio F. Tormena, Mario J. A. Saad Proceedings of the National Academy of Sciences of the United States of America, 2023
Prognostic value of TERF1 expression in prostate cancer Gabriel Arantes dos Santos, Nayara Izabel Viana, Ruan Pimenta, Vanessa Ribeiro Guimarães, Juliana Alves de Camargo, Poliana Romão, Sabrina T. Reis, Katia Ramos Moreira Leite, Miguel Srougi Journal of the Egyptian National Cancer Institute, 2021
Shorter leukocyte telomere length is associated with severity of COVID-19 infection. Gabriel Arantes dos Santos, Ruan Pimenta, Nayara I. Viana, Vanessa R. Guimarães, Poliana Romão, Patrícia Candido, Juliana A. de Camargo, Diná M. Hatanaka, Paula GS. Queiroz, Alexandre Teruya, Katia R.M. Leite, Victor Srougi, Miguel Srougi, Sabrina T. Reis Biochemistry and Biophysics Reports, 2021
Exercise activates the hypothalamic S1PR1–STAT3 axis through the central action of interleukin 6 in mice Vagner R. R. Silva, Thayana O. Micheletti, Carlos K. Katashima, Luciene Lenhare, Joseane Morari, Alexandre Moura‐Assis, José C. de Lima‐Júnior, Juliana A. Camargo, Gabriela R. Passos, Rodrigo S. Gaspar, Licio A. Velloso, Mario J. Saad, Adelino S. R. da Silva, Leandro P. Moura, Dennys E. Cintra, José R. Pauli, Eduardo R. Ropelle Journal of Cellular Physiology, 2018
