cyclin-d1 and Prostatic-Neoplasms--Castration-Resistant

Aberrant activation of several signaling pathways has been implicated in prostate cancer (PCa) progression to castrate-resistant prostate cancer (CRPC). Phosphoinositide-3-kinase/Protein Kinase B/mechanistic Target of Rapamycin (PI3K/AKT/mTOR) and Hedgehog/GLI (Hh/GLI) pathways are major participants in progression to CRPC. In this sense, the current work aims to assess the potential antitumor effects resulting from co-targeting the aforementioned pathways in PC3 cells with Dactolisib as a dual PI3K/mTOR inhibitor and GANT61 as a GLI1 antagonist. Three replica of PC3 cells were assigned for four treatment groups; vehicle control, Dactolisib-treated, GANT61-treated, and combination-treated groups. GLI1 gene expression was determined by quantitative real-time PCR while active caspase-3 was determined colorimetrically. P-AKT, p70 ribosomal s6 protein kinase 1 (pS6K1), cyclin D1, vascular endothelial growth factor 1 (VEGF1), and Microtubule-associated proteins 1A/1B light chain 3 (LC3) protein levels were determined by ELISA technique. GLI1 gene expression was down-regulated as a result of Dactolisib, GANT61, and their combination. Additionally, both drugs significantly reduced p-AKT, pS6K1, cyclin D1, and VEGF1 protein levels. Dactolisib elevated LC3 protein levels and GANT61 augmented Dactolisib effect on LC3. Moreover, only Dactolisib/GANT61combination significantly increased active caspase-3 level. To sum up, Dactolisib/GANT61 combination was shown to be promising in PCa treatment. Further in-vitro and in-vivo studies are warranted to support our findings.

Topics: Caspase 3; Cell Line, Tumor; Cyclin D1; Hedgehog Proteins; Humans; Imidazoles; Male; Phosphatidylinositol 3-Kinases; Prostatic Neoplasms, Castration-Resistant; Proto-Oncogene Proteins c-akt; Pyridines; Pyrimidines; Quinolines; TOR Serine-Threonine Kinases; Vascular Endothelial Growth Factor A; Zinc Finger Protein GLI1

Two androgen receptor (AR)-targeted therapies, enzalutamide and abiraterone acetate plus prednisone (abiraterone), have been approved for the treatment of metastatic castration-resistant prostate cancer (CRPC). Many patients respond to these agents, but both de novo and acquired resistance are common. The authors characterized resistant phenotypes that emerge after treatment with abiraterone or enzalutamide.. Patients who received abiraterone or enzalutamide in the course of routine clinical care were consented for serial blood collection. A proprietary system (CellSearch) was used to enumerate and enrich circulating tumor cells (CTCs). RNA-sequencing (RNA-seq) was performed on pools of up to 10 epithelial cell adhesion molecule (EpCAM)-positive/CD45-negative CTCs. The impact of gene expression changes observed in CTCs between patients who responded or were resistant to abiraterone/enzalutamide therapies was further explored in a model cell line system.. RNA-seq data from CTCs identified mutations commonly associated with CRPC as well as novel mutations, including several in the ligand-binding domain of AR that could facilitate escape from AR-targeted agents. Ingenuity pathway analysis of differentially regulated genes identified the transforming growth factor β (TGFβ) and cyclin D1 (CCND1) signaling pathways as significantly upregulated in drug-resistant CTCs. Transfection experiments using enzalutamide-sensitive and enzalutamide-resistant LNCaP cells confirmed the involvement of SMAD family member 3, a key mediator of the TGFβ pathway, and of CCND1 in resistance to enzalutamide treatment.. The current results indicate that RNA-seq of CTCs representing abiraterone and enzalutamide sensitive and resistant states can identify potential mechanisms of resistance. Therapies targeting the downstream signaling mediated by SMAD family member 3 (SMAD3) and CCND1, such as cyclin-dependent kinase 4/cyclin-dependent kinase 6 inhibitors, could provide new therapeutic options for the treatment of antiandrogen-resistant disease. Cancer 2018;124:1216-24. © 2017 American Cancer Society.

Topics: Abiraterone Acetate; Aged; Aged, 80 and over; Androgen Antagonists; Antineoplastic Combined Chemotherapy Protocols; Benzamides; Biomarkers, Tumor; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; Drug Resistance, Neoplasm; Humans; Male; Middle Aged; Neoplastic Cells, Circulating; Nitriles; Phenylthiohydantoin; Prostatic Neoplasms, Castration-Resistant; Protein Kinase Inhibitors; Signal Transduction; Smad3 Protein

To explore the inhibitory effect of genistein (GEN) on the proliferation of VCaP castration-resistant prostate cancer (CRPC) cells.. VCaP CRPC cells were treated with GEN at the concentrations of 0, 12.5, 25, 50, 100, and 200 μmol/L for 24, 48, and 72 hours followed by determination of their proliferation by CCK-8 assay and their cycle by flow cytometry. The expression of Ki-67 in the cells was detected by immunocytochemistry and the levels of PSA, Cyclin D1, PCNA, and P53 determined by Western blot.. After 72 hours of treatment with GEN at 12.5, 25, 50, 100, and 200 μmol/L, the inhibition rates of the VCaP cells were (25.38±0.02)%, (31.14±0.29)%, (45.27±0.03)%, (52.19±0.05)%, and (68.21±0.19)%, respectively, all significantly higher than in the 0 μmol/L group (［10.08±0.02］%)(P<0.05). GEN caused the arrest of the VCaP cells in the G2/M phase (P<0.05) and inhibited the expression of Ki-67. The expressions of PSA, Cyclin D1, and PCNA were gradually down-regulated while that of P53 up-regulated with the increased concentration of GEN (P<0.05).. GEN inhibits the proliferation of VCaP CRPC cells by arresting the cell cycle with related protein expression changes.. 目的： 研究染料木黄酮(GDN)对去势抵抗性前列腺癌细胞VCaP增殖的抑制作用及可能机制。方法： 以不同浓度（0、12.5、25、50、100、200 μmol/L）的GEN处理去势抵抗性前列腺癌细胞VCaP，分别在24、48、72 h以CCK-8法测定细胞增殖；以流式细胞术检测细胞周期；以免疫荧光法检测Ki-67表达水平；以Western 印迹法检测Cyclin D1、PCNA、P53及PSA表达水平。结果： 12.5、25、50、100、200 μmol/L GEN作用于VCaP细胞72 h后，对VCaP细胞的抑制率分别为（25.38±0.02）%、（31.14±0.29）%、（45.27±0.03）%、（52.19±0.05）%与（68.21±0.19）%，与对照组［(10.08±0.02)%］相比差异有统计学意义（P<0.05）。流式细胞术显示GEN可导致VCaP细胞G2/M期阻滞（P<0.05）。免疫细胞化学法检测显示GEN能够抑制细胞中Ki-67的表达。Western印迹显示去势抵抗性前列腺癌细胞内PSA、Cyclin D1、PCNA随着GEN浓度的增加逐渐下调，P53随着GEN浓度的增加逐渐上调(P<0.05)。 结论： GEN能够抑制体外培养的去势抵抗性前列腺癌细胞VCaP增殖，其作用机制可能与阻滞细胞周期并伴随相关周期蛋白表达的改变有关。.

Topics: Cell Count; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Genistein; Humans; Male; Proliferating Cell Nuclear Antigen; Prostate-Specific Antigen; Prostatic Neoplasms, Castration-Resistant

Patients with hormone-resistant prostate cancer (PCa) have higher biochemical failure rates following radiation therapy (RT). Cyclin D1 deregulated expression in PCa is associated with a more aggressive disease: however its role in radioresistance has not been determined. Cyclin D1 levels in the androgen-independent PC3 and 22Rv1 PCa cells were stably inhibited by infecting with cyclin D1-shRNA. Tumorigenicity and radiosensitivity were investigated using in vitro and in vivo experimental assays. Cyclin D1 silencing interfered with PCa oncogenic phenotype by inducing growth arrest in the G1 phase of cell cycle and reducing soft agar colony formation, migration, invasion in vitro and tumor formation and neo-angiogenesis in vivo. Depletion of cyclin D1 significantly radiosensitizes PCa cells by increasing the RT-induced DNA damages by affecting the NHEJ and HR pathways responsible of the DNA double-strand break repair. Following treatment of cells with RT the abundance of a biomarker of DNA damage, γ-H2AX, was dramatically increased in sh-cyclin D1 treated cells compared to shRNA control. Concordant with these observations DNA-PKcs-activation and RAD51-accumulation, part of the DNA double-strand break repair machinery, were reduced in shRNA-cyclin D1 treated cells compared to shRNA control. We further demonstrate the physical interaction between CCND1 with activated-ATM, -DNA-PKcs and RAD51 is enhanced by RT. Finally, siRNA-mediated silencing experiments indicated DNA-PKcs and RAD51 are downstream targets of CCND1-mediated PCa cells radioresistance. In summary, these observations suggest that CCND1 is a key mediator of PCa radioresistance and could represent a potential target for radioresistant hormone-resistant PCa.

Topics: Animals; Apoptosis; Blotting, Western; Cell Adhesion; Cell Movement; Cell Proliferation; Cyclin D1; DNA Breaks, Double-Stranded; DNA Repair; Fluorescent Antibody Technique; Histones; Humans; Immunoenzyme Techniques; Male; Mice; Mice, Nude; Phosphorylation; Prostatic Neoplasms, Castration-Resistant; Radiation Tolerance; Radiation-Sensitizing Agents; RNA, Small Interfering; Signal Transduction; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

To explore the antitumoral effect of indirubin on androgen-independent prostate cancer PC-3 cells and its possible mechanisms.. We measured the inhibitory effect of indirubin on the proliferation of prostate cancer PC-3 cells using MTT assay, detected their cell cycles by flow cytometry, and determined the expressions of the cell cycle regulatory protein cyclin D1 and its related downstream gene c-myc by Western blot.. The viability of the PC-3 cells was significantly decreased by indirubin in a concentration-dependent manner, reduced to 52. 2% and 13. 6% at 5 and 10 µmol/L, respectively. The cell cycle of the PC-3 cells was markedly inhibited by indirubin at 5 µmol/L, with the cells remarkably increased in the G0 and G1 phases and decreased in the S and G2/M phases. Meanwhile, indirubin also inhibited the expressions of cyclin D1 and c-myc in the Wnt signaling pathway.. Indirubin can suppress the proliferation of androgen-independent prostate cancer PC-3 cells, which may be associated with its inhibitory effect on the cell cycle and Wnt signaling pathway.

Topics: Antibiotics, Antineoplastic; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Coloring Agents; Cyclin D1; Dose-Response Relationship, Drug; Genes, myc; Humans; Indoles; Male; Prostatic Neoplasms, Castration-Resistant; Proto-Oncogene Proteins c-myc; Tetrazolium Salts; Thiazoles

Prostate cancer (PCa) is the second leading cause of cancer-related deaths in men. Studies show that consumption of polyunsaturated fatty acids (PUFA) modulates the development and progression of prostate cancer. High amounts of omega-6 fatty acids have been linked with increased prostate cancer risk, whereas omega-3 fatty acids have been shown to inhibit PCa growth. However, because omega-3 and omega-6 are both essential fatty acids and part of a complete diet, it is more relevant to determine the ideal ratio of the two that would allow patients to benefit from the therapeutic properties of omega-3 fatty acids. LNCaP prostate cancer cells were treated with dietary-based ratios of omega-6 to omega-3 fatty acids under hormone-deprivation conditions, and effects on various cellular processes were determined. A low omega-6 to omega-3 PUFA ratio can delay the progression of cells toward castration-resistance by suppressing pathways involved in prostate cancer progression, such as the Akt/mTOR/NFκB axis. It also suppresses the expression of cyclin D1, and activation of caspase-3 and annexin V staining shows induction of proapoptotic events. Taken together, our data demonstrates that maintaining a low omega-6 to omega-3 fatty acids ratio can enhance efficacy of hormone ablation therapy.

Topics: Apoptosis; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclin D1; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Humans; Male; NF-kappa B; Phosphatidylinositol 3-Kinases; Prostatic Neoplasms; Prostatic Neoplasms, Castration-Resistant; Signal Transduction; TOR Serine-Threonine Kinases