Tetrandrine suppresses metastatic phenotype of prostate cancer cells by regulating Akt/mTOR/MMP-9 signaling pathway
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Prostate cancer is one of the leading causes of cancer deaths among men. New gene expressed in prostate (NGEP), is a prostate-specific gene expressed only in normal prostate and prostate cancer tissue. Because of its selective expression in prostate cancer cell surface, NGEP is a potential immunotherapeutic target. To target the NGEP in prostate cancer, it is essential to investigate its expression in prostate cancer cells.In the present study, we investigated NGEP expression in LNCaP and DU145 cells by real time and RT-PCR, flow cytometric and immunocytochemical analyses.Real time and RT-PCR analyses of NGEP expression showed that NGEP was expressed in the LNCaP cells but not in DU145 cells. The detection of NGEP protein by flow cytometric and immunocytochemistry analyses indicated that NGEP protein was weakly expressed only in LNCaP cell membrane.Our results demonstrate that LNCaP cell line is more suitable than DU145 for NGEP expression studies; however, its low-level expression is a limiting issue. NGEP expression may be increased by androgen supplementation of LNCaP cell culture medium.
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1144 Objectives FDG PET can be limited in the evaluation of prostate cancer. Several other PET tracers show promise. Using whole genome microarray profile of cell lines, we compared the expression of messages related to the key pathways linked to radiotracer uptake and retention to inform selection and development of prostate cancer imaging radiotracers. Methods Total RNA from PC3, DU145, LnCaP prostate cancer cell lines and duplicated immortal prostate cell line were extracted. Affy U133 plus 2.0 genomic chips (47,000 transcripts) were used in this study and the raw data were normalized and then analyzed with Robust Multichip Average (RMA) and Spotfire software. Results The rank of folate hydrolase expression from high to low expression as compared to normal prostate cell line was LnCaP (13909.6%), DU145 (107.9%), and PC3 (90.6%). PC3 presented minimally higher HK1 (129.4%) and PKM2 (104.5%) expression and DU145 showed higher PKM2 (166.7%) as compared to normal. The ranks of thymidine kinase (TK1) and choline kinase (CHKA) from high to low expression as compared to normal prostate cell line were PC3 (1104.7%), DU145 (934.4%), and LnCaP (727.7%) in TK1 and PC3 (383.9%), LnCaP (223.9%), and DU145 (160.7%) in CHKA. Only LnCaP (146.3%) presents slightly higher expression in acetoacetyl-CoA synthetase (AACS). Conclusions All of three prostate metastatic cell lines in this study presented with moderate to marked overexpression of TK1 and moderate overexpresion of CHKA, which suggested that FLT and Choline may use for detection of hematogeneous and lymphogeneous metastases. LNCaP presented with marked overexpression of FOLH1, but was not overexpressed in two non nodal lines. Glycolysis and acetate metabolism pathways were not impressively elevated in this study except mild to moderate overexpression of PKM2 in PC3 and DU145 and AACS in LNCaP
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Objective To investigate the role of solanine in human androgen-dependent prostate cancer cell line LNCaP and androgen-independent prostate cancer cell Du145.Methods The influence on the proliferation of LNCaP and Du145 cells were evaluated by MTT assay.Cell cycle and cell apoptosis of LNCaP and Du145 were assessed by flow cytometry and fluorescence microscope.The protein of expressions of Bcl-2,Bax,and Caspase-3 were determined by Western blot.Results Solanine could significantly inhibit the proliferation of the two cells in time-and dose-dependent manners(P0.05);also induced S-phase arrest of LNCaP and Du145 cells.The apoptosis ratio in treatment groups were significantly higher than that in 0 μg/mL group,with early apoptosis.Significant down-regulation of Bcl-2 and Caspase-3 was induced by solanine,with no effect on Bax protein.Conclusion Solanine provides anti-prostate cancer effect by inhibiting cell proliferation,S-phase arresting,and inducing apoptosis.
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Abstract [Background] The mechanisms that prostate cancer (PCa) progresses to castration-resistant PCa (CRPC) are adaptation and clonal selection. We hypothesized that PCa proliferated while each mechanism cooperated when PCa recurred. [Materials and Methods] After LNCaP cells were transfected with luciferase reporter driven by PSA promoter, LNCaP were cocultured with androgen-insensitive DU145 or PC-3. Then DHEA or DHT were added to the medium and luciferase assay was performed. Also, we added DHEA or DHT in the medium and measured various androgens level by LC-MS/MS. As for proliferation, LNCaP were cocultured with DU145 or PC -3 in 2-layer chamber and number of LNCaP were counted. DU145 and PC-3 were also cocultured with LNCaP cells for 4 days. We also checked migration by using 2-layer chamber and migrated cells were counted. [Results] Whereas DHEA was converted into DHT in DU145 and induced PSA promoter activity in LNCaP, the effect was not found in PC-3. Moreover, DU145 elevated DHT-induced PSA promoter activity. DU145 promoted LNCaP proliferation stimulated by DHT and DHEA, but PC-3 did not. LNCaP also promoted proliferation of DU145 and PC-3. LNCaP promoted migration of PC-3, but not DU145. [Conclusion] Cross-talk between androgen-sensitive PCa cells and androgen-insensitive PCa cells might regulate progression of CRPC. Citation Format: Yuta Takezawa, Atsushi Mizokami, Kazuaki Machioka, Kouji Izumi, Hiroaki Iwamoto, Maolake Aerken, Natsagdorg Ariunbold, Mikio Namiki. Crosstalk of androgen sensitive prostate cancer cells and insensitive prostate cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1581.
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A series of novel arylpiperazine derivatives was synthesized. The in vitro cytotoxic activities of all synthesized compounds against three human prostate cancer cell lines (PC-3, LNCaP, and DU145) were evaluated by a CCK-8 assay. Compounds 9 and 15 exhibited strong cytotoxic activities against LNCaP cells (IC50<5 μM), and compound 8 (IC50=8.25 μM) possessed the most potent activity against DU145 cells. However, these compounds also exhibited cytotoxicity towards human epithelial prostate normal cells RWPE-1. The structure-activity relationship (SAR) of these arylpiperazine derivatives was also discussed based on the obtained experimental data.
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To observe the expression of Coxsackie B virus-adenovirus receptor (CAR) in two prostate cancer cell lines with different metastatic potentials.The expressions of CAR in two prostate cancer cell lines (Du145 and LNCaP) with different metastatic potentials were detected by Western blotting. The Transwell polycarbonate filter was used to analyze the invasive potency.As one of the adhesion associated proteins, CAR highly expressed in the LNCaP cell line, which is well known with a low metastatic potential, and lowly expressed in Du145 with a high metastatic potential (P < 0.01). The invasive potency of Du145 was significantly higher than that of LNCaP (P < 0.05).There was a difference in the metastatic phenotypes of CAR among cell lines with different metastatic potentials. The expressions of CAR proteins may play an important role in repressing the metastasis of prostate cancer.
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