Abstract Objective The present study aimed to investigate FOXO3a deregulation in Uterine Smooth Muscle Tumors (USMT) and its potential association with cancer development and prognosis. Methods The authors analyzed gene and protein expression profiles of FOXO3a in 56 uterine Leiomyosarcomas (LMS), 119 leiomyomas (comprising conventional and unusual leiomyomas), and 20 Myometrium (MM) samples. The authors used techniques such as Immunohistochemistry (IHC), FISH/CISH, and qRT-PCR for the present analyses. Additionally, the authors conducted an in-silico analysis to understand the interaction network involving FOXO3a and its correlated genes. Results This investigation revealed distinct expression patterns of the FOXO3a gene and protein, including both normal and phosphorylated forms. Expression levels were notably elevated in LMS, and Unusual Leiomyomas (ULM) compared to conventional Leiomyomas (LM) and Myometrium (MM) samples. This upregulation was significantly associated with metastasis and Overall Survival (OS) in LMS patients. Intriguingly, FOXO3a deregulation did not seem to be influenced by EGF/HER-2 signaling, as there were minimal levels of EGF and VEGF expression detected, and HER-2 and EGFR were negative in the analyzed samples. In the examination of miRNAs, the authors observed upregulation of miR-96-5p and miR-155-5p, which are known negative regulators of FOXO3a, in LMS samples. Conversely, the tumor suppressor miR-let7c-5p was downregulated. Conclusions In summary, the outcomes of the present study suggest that the imbalance in FOXO3a within Uterine Smooth Muscle Tumors might arise from both protein phosphorylation and miRNA activity. FOXO3a could emerge as a promising therapeutic target for individuals with Unusual Leiomyomas and Leiomyosarcomas (ULM and LMS), offering novel directions for treatment strategies. FOXOa FOXO USMT (USMT prognosis 5 , (LMS) 11 comprising leiomyomas, leiomyomas) 2 MM (MM samples IHC, IHC (IHC) FISHCISH FISH CISH FISH/CISH qRTPCR qRT PCR analyses Additionally insilico silico genes forms ULM LM (LM OS (OS patients Intriguingly EGF/HER2 EGFHER2 EGFHER EGF/HER HER EGF/HER- signaling detected HER2 HER- miRNAs miR965p miRp miR 96 5p p miR1555p, miR1555p 155 5p, miR-155-5p Conversely miRlet7c5p miRletcp let7c let c downregulated summary activity LMS) strategies (LMS 1 (IHC 9 15 miRlet letc

Abstract Objective To evaluate the effects of estrogen, raloxifene and genistein on the expression of KISS1 (kisspeptin), KISS1R (kisspeptin receptor), AR (androgen receptor) and INSR (insulin receptor) in the bones of ovariectomized rats. Methods Forty-eight adult rats were randomly divided into 6 groups, containing 8 animals each: G1–nonovariectomized control; G2–ovariectomized and treated with conjugated equine estrogens (50 µg/Kg/day); G3–ovariectomized and treated with raloxifene (0.75 mg/kg/day); G4–ovariectomized animal that received soy extract with genistein (300 mg/kg/day); G5–ovariectomized animal that received estrogen and genistein; and G6–ovariectomized animal that received estrogen and raloxifene. Three months after surgery, the castrated animals received the drugs orally daily for 120 days. All animals were sacrificed after this period, by deepening the anesthesia. The left tibia was removed for total RNA extraction and analysis of gene expression of KISS1, KISS1R, AR and INSR, by quantitative real-time polymerase chain reaction (qRT-PCR). Results KISS1 was not detected in any of the treated groups. KISS1R, INSR and AR showed higher expression in the G3 group (p < 0.001), while lower levels of transcripts for these genes were observed in G4 and G5. G2 animals showed hypoexpression of the evaluated genes. Conclusion The results indicate that raloxifene, alone or combined with estrogen, was able to induce the expression of genes associated with the recovery of bone tissue homeostasis in ovariectomized rats. KISS kisspeptin, kisspeptin , (kisspeptin) KISSR R receptor, receptor androgen insulin Fortyeight Forty eight groups each G1nonovariectomized Gnonovariectomized G1 nonovariectomized G control G2ovariectomized Govariectomized 50 (5 µg/Kg/day µgKgday µg Kg day µg/Kg/day) G3ovariectomized 0.75 075 0 75 (0.7 mg/kg/day mgkgday mg kg mg/kg/day) G4ovariectomized 300 (30 G5ovariectomized G5 G6ovariectomized G6 surgery 12 days period anesthesia realtime real time qRTPCR. qRTPCR qRT PCR . (qRT-PCR) p 0.001, 0001 0.001 001 0.001) 5 ( 0.7 07 7 (0. 30 (3 1 (qRT-PCR 000 0.00 00 0. (0 3 0.0

Resumo Objetivo Avaliar os efeitos do estrogênio, raloxifeno e genisteína na expressão de KISS1 (kisspeptina), KISS1R (receptor da kisspeptina), AR (receptor de androgênio) e INSR (receptor de insulina) nos ossos de ratas ovariectomizadas. Métodos Quarenta e oito ratas adultas foram divididas aleatoriamente em 6 grupos, contendo 8 animais cada: G1–controle não ovariectomizado); G2–ovariectomizado e tratado com estrogênios conjugados equinos (50 µg/Kg/dia); G3–ovariectomizado e tratado com raloxifeno (0,75 mg/kg/dia); G4–ovariectomizado que recebeu extrato de soja com genisteína (300 mg/kg/dia); G5–ovariectomizado que recebeu estrogênio e genisteína; e G6–ovariectomizado que recebeu estrogênio e raloxifeno. Após 3 meses da cirurgia, os animais castrados receberam os fármacos diariamente por via oral, durante 120 dias. Todos os animais foram sacrificados após esse período, por aprofundamento da anestesia. A tíbia esquerda foi removida para extração de RNA total e análise da expressão gênica de KISS1, KISS1R, AR e INSR, por reação de cadeia de polimerase quantitativa em tempo real (quantitative real-time polymerase chain reaction, qRT-PCR, em inglês). Resultados KISS1 não foi detectado em nenhum dos grupos tratados. KISS1R, INSR e AR mostraram maior expressão no grupo G3 (p < 0,001), enquanto menores níveis de transcritos para esses genes foram observados em G4 e G5. Os animais de G2 apresentaram hipoexpressão dos genes avaliados. Conclusão Os resultados indicam que o raloxifeno, isolado ou combinado com estrogênio, foi capaz de induzir a expressão de genes associados à recuperação da homeostase do tecido ósseo em ratas ovariectomizadas. KISS kisspeptina, kisspeptina , (kisspeptina) KISSR R receptor kisspeptina) androgênio insulina ovariectomizadas cada G1controle Gcontrole G1 controle G ovariectomizado ovariectomizado) G2ovariectomizado Govariectomizado 50 (5 µg/Kg/dia µgKgdia µg Kg dia µg/Kg/dia) G3ovariectomizado 0,75 075 0 75 (0,7 mg/kg/dia mgkgdia mg kg mg/kg/dia) G4ovariectomizado 300 (30 G5ovariectomizado G5 G6ovariectomizado G6 cirurgia oral 12 dias período anestesia quantitative realtime time reaction qRTPCR, qRTPCR qRT PCR, PCR qRT-PCR inglês. inglês . inglês) tratados p 0,001, 0001 0,001 001 0,001) avaliados (kisspeptina 5 ( 0,7 07 7 (0, 30 (3 1 000 0,00 00 0, (0 0,0

Abstract FOXO3a dysregulation is frequently implicated in tumorigenesis, and its inhibition can occur by several molecular mechanisms. Among these, post-transcriptional suppression by miRNAs has been associated with various cancers initiation. Here, we assessed the expression profiles of the most relevant miRNAs for breast tumorigenesis, using Luminal A (LA) and Triple-Negative (TN) breast cancer from Brazilian patients, by the quantitative real time-PCR method. Their potential prognostic role for the patients was also evaluated. We identified the miRNAs miR-96-5p and miR-182-5p, de-scribed as negative regulators of FOXO3A, with differential expression both in LA and TN tumors when compared to normal tissue. The miR-96-5p and miR-182-5p miRNAs were upregulated in LA (7.82 times, p < 0.005; 6.12 times, p < 0.005, respectively) and TN breast cancer samples (9.42 times, p < 0.0001; 8.51 times, p < 0.0001) compared to normal tissues. The samples with higher miR-96-5p and miR-182-5p expression (FR ≥ 4) were submitted for FOXO3a immunostaining. Reduced protein detection was observed in all of the tumors compared to normal tissues. The most prominent miRNA expression and FOXO3a protein suppression were observed in TN samples (p < 0.001), indicating the relevant role of these molecules in this tumor biology and clinical behavior. Our results corroborate the literature regarding to the relevance of FOXO3a in the breast cancer, and they open new perspectives for alternative target therapy options for Brazilian patients expressing both FOXO3a and its regulatory miRNAs. FOXOa FOXO a tumorigenesis mechanisms posttranscriptional post transcriptional initiation Here (LA TripleNegative Triple Negative (TN timePCR time PCR method evaluated miR965p miRp miR 96 5p miR1825p, miR1825p 182 5p, described de scribed FOXO3A FOXOA tissue 7.82 782 7 82 (7.8 times 0.005 0005 0 005 612 6 12 6.1 respectively 9.42 942 9 42 (9.4 0.0001 00001 0001 851 8 51 8.5 tissues FR 4 immunostaining 0.001, 0.001 , 001 0.001) behavior 18 7.8 78 (7. 0.00 000 00 61 1 6. 9.4 94 (9. 0.000 0000 85 5 8. 7. (7 0.0 9. (9 ( 0.

OBJECTIVES: The present study aimed to contribute to the catalog of genetic mutations involved in the carcinogenic processes of uterine sarcomas (USs) and carcinosarcomas (UCSs), which may assist in the accurate diagnosis of, and selection of treatment regimens for, these conditions. METHODS: We performed gene-targeted next-generation sequencing (NGS) of 409 cancer-related genes in 15 US (7 uterine leiomyosarcoma [ULMS], 7 endometrial stromal sarcoma [ESS], 1 adenosarcoma [ADS]), 5 UCS, and 3 uterine leiomyoma (ULM) samples. Quality, frequency, and functional filters were applied to select putative somatic variants. RESULTS: Among the 23 samples evaluated in this study, 42 loss-of-function (LOF) mutations and 111 missense mutations were detected, with a total of 153 mutations. Among them, 66 mutations were observed in the Catalogue of Somatic Mutations in Cancer (COSMIC) database. TP53 (48%), ATM (22%), and PIK3CA (17%) were the most frequently mutated genes. With respect to specific tumor subtypes, ESS showed mutations in the PDE4DIP, IGTA10, and DST genes, UCS exhibited mutations in ERBB4, and ULMS showed exclusive alterations in NOTCH2 and HER2. Mutations in the KMT2A gene were observed exclusively in ULM and ULMS. In silico pathway analyses demonstrated that many genes mutated in ULMS and ESS have functions associated with the cellular response to hypoxia and cellular response to peptide hormone stimulus. In UCS and ADS, genes with most alterations have functions associated with phosphatidylinositol kinase activity and glycerophospholipid metabolic process. CONCLUSION: This preliminary study observed pathogenic mutations in US and UCS samples. Further studies with a larger cohort and functional analyses will foster the development of a precision medicine-based approach for the treatment of US and UCS.