RESUMO
Prostate cancer (PCa) is a prevalent malignancy among men, with a majority of patients presenting with distant metastases at the time of initial diagnosis. These patients are at a heightened risk of developing more aggressive castrationresistant PCa following androgen deprivation therapy, which poses a greater challenge for treatment. Notably, the inhibition of tumor angiogenesis should not be considered an ineffective treatment strategy. The regulatory role of CDK12 in transcriptional and posttranscriptional processes is essential for the proper functioning of various cellular processes. In the present study, the expression of CDK12 was first knocked down in cells using CRISPR or siRNA technology. Subsequently, RNAseq analysis, coimmunoprecipitation, western blotting, reverse transcriptionquantitative polymerase chain reaction and the LinkedOmics database were employed to reveal that CDK12 inhibits insulin like growth factor binding protein 3 (IGFBP3). Western blot analysis also demonstrated that CDK12 promoted VEGFA expression by inhibiting IGFBP3, which involves the Akt signaling pathway. Then, CDK12 was found to promote PCa cell proliferation, cell migration and angiogenesis by inhibiting IGFBP3 through cell proliferation assays, cell migration assays and tube formation assays, respectively. Finally, animal experiments were performed for in vivo validation. It was concluded that CDK12 promoted PCa and its angiogenesis by inhibiting IGFBP3.
Assuntos
Neoplasias da Próstata , Animais , Masculino , Humanos , Neoplasias da Próstata/genética , Antagonistas de Androgênios , Angiogênese , Bioensaio , Quinases Ciclina-Dependentes , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genéticaRESUMO
Although patients with early localized prostate cancer can survive longer, castration-resistant prostate cancer (CRPC) has gradually emerged with the use of androgen deprivation therapy (ADT). N-Myc and TEM8 play a vital role in the progression of several cancer types. However, the underlying mechanism of how N-Myc and TEM8 promote the progression of prostate cancer remains unclear. In this study, the expression of N-Myc and TEM8 was detected in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) tissues by immunohistochemistry (IHC). LNCaP cell lines were maintained in RPMI 1640 medium supplemented with 10% charcoal-stripped fetal bovine serum. Subsequently, R language software was used to verify our results. Tubule formation assay of human umbilical vein endothelial cell (HUVEC) was conducted to examine the effect of N-Myc and TEM8 overexpression on angiogenesis in prostate cancer cells. IHC results showed a positive correlation between the expression of N-Myc and TEM8 in prostate cancer tissues. Further analysis showed that N-Myc and TEM8 were associated with clinicopathological features and poor prognosis in prostate cancer patients. Moreover, the overexpression of N-Myc and TEM8 promoted proliferation of prostate cancer cells and angiogenesis. Additionally, N-Myc and TEM8 overexpression was associated with therapeutic resistance. We further found that N-Myc promoted angiogenesis and therapeutic resistance in prostate cancer via TEM8. Hence, targeting N-Myc/TEM8 pathway in prostate cancer would be a novel therapeutic strategy to enhance the treatment of prostate cancer patients.
Assuntos
Antagonistas de Androgênios/farmacologia , Resistencia a Medicamentos Antineoplásicos , Proteínas dos Microfilamentos/metabolismo , Proteína Proto-Oncogênica N-Myc/metabolismo , Neovascularização Patológica/patologia , Hiperplasia Prostática/patologia , Neoplasias de Próstata Resistentes à Castração/patologia , Receptores de Superfície Celular/metabolismo , Idoso , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Proteínas dos Microfilamentos/genética , Proteína Proto-Oncogênica N-Myc/genética , Prognóstico , Hiperplasia Prostática/tratamento farmacológico , Hiperplasia Prostática/metabolismo , Neoplasias de Próstata Resistentes à Castração/irrigação sanguínea , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/metabolismo , Receptores Androgênicos/química , Receptores de Superfície Celular/genética , Taxa de Sobrevida , Células Tumorais CultivadasRESUMO
The oncoprotein NMyc has a carcinogenic effect in numerous types of cancer, and it can cause castration resistance in prostate cancer (PCa), and leads to the development of small cell neuroendocrine cancer by regulating multiple target genes. Immunohistochemical staining, RTqPCR, western blotting, wound healing and CCK8 assays were used to detect the expression of NMyc and FSCN1 as well as AR and CgA at the human level and cell level. The immunohistochemical results revealed that the protein levels of NMyc protooncogene protein (NMyc) and fascin (FSCN1) in PCa were significantly higher than that of hyperplastic tissues (P<0.05), and there was a weak correlation between them (P=0.002). In vitro, NMyc and FSCN1 were overexpressed in LNCaP and C42 cell lines. The results revealed the promoting effect of NMyc and FSCN1 on malignant progression of PCa. In addition, the endogenous FSCN1 was knocked down in the C42 cell line, and the results revealed that the silencing of FSCN1 enhanced the expression of NMyc and weakened the expression of the neuroendocrine marker CgA. Therefore, the present findings indicated that NMyc may promote the malignant process of PCa by regulating FSCN1 and FSCN1 may have a reverse regulatory effect on NMyc.