A new HSP90 inhibitor as therapeutic agent for bladder cancer.

2017 ◽  
Vol 35 (6_suppl) ◽  
pp. 416-416
Author(s):  
Weiya Liu ◽  
Eugene K. Lee ◽  
Karim Pirani ◽  
Brian S. J. Blagg ◽  
Jeffrey M. Holzbeierlein
Keyword(s):  
Bladder Cancer ◽  
Heat Shock ◽  
Western Blot ◽  
Protein Degradation ◽  
Cancer Cells ◽  
Hsp90 Inhibitor ◽  
Client Protein ◽  
Antiproliferative Effects ◽  
Bladder Cancer Cells ◽  
Shock Response

416 Background: Hsp90 represents one of the most promising biological targets for the treatment of cancer, including bladder cancer. A number of Hsp90 inhibitors that target the N-terminal ATP-binding pocket have demonstrated potent antiproliferative effects. However, a major drawback is that they induce a prosurvival heat shock response (HSR). We demonstrate the effects of a novel Hsp90 beta selective inhibitor on bladder cancer cells, which shows potent antiproliferative effects without inducing HSR. Methods: Cell Titer-Glo luminescent anti-proliferative assay was used to determine the IC50 numbers in UC3 cells. Trypan Blue Cytotoxicity assay was performed for 24h treatment with increasing concentrations of the inhibitor. Effects of the cmpound on Hsp90’s client protein degradation were investigated by Western Blot. Results: This new compound exhibits potent anti-proliferative in bladder cancer cells. IC50 number is determined as 0.30 µM for UC3 cancer cells. The toxicity assay was also performed over UC3 cells at 24h.1uM KU new compound has the similar effects on UC3 cells as 10 uM 17AAG: inhibit the cancer cells growth to half, but maintain over 60% viability of the cells. The western blot were also performed over UC3 cells, and some new target proteins such as FGFR3 and PKM2 were investigated. The data showed that, this new compound would not induce the heat shock response like 17AAG (Hsp27), and did cause some Hsp90β related protein degradation (CXCR4). FGFR3, PKM2, Her2, Hsf-1and B-raf all show degradation to different extent. Conclusions: A novel Hsp90 inhibitor, exhibits potent anti-proliferative and cytotoxic activity along with client protein degradation, without induction of HSR in bladder cancer cell lines. The reduction of Hsp90 beta related client protein caused by this compound suggests the potential to develop isoform specific inhibitors of Hsp90 for better antitumor therapies.

New HSP90 selective inhibitors as therapeutic agents for prostate and bladder cancer.

2018 ◽  
Vol 36 (6_suppl) ◽  
pp. 285-285 ◽  
Author(s):  
Weiya Liu ◽  
Derek Jensen ◽  
Eugene Lee ◽  
Jessie Gills ◽  
Jeffrey M. Holzbeierlein
Keyword(s):  
Bladder Cancer ◽  
Western Blot ◽  
Protein Degradation ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Selective Inhibition ◽  
Client Protein ◽  
Selective Inhibitors ◽  
Oncogenic Pathways ◽  
Bladder Cancer Cells

285 Background: Hsp90 is a molecular chaperone responsible for folding many of the proteins directly associated with cancer progression and consequently, inhibition of the Hsp90 protein folding machinery results in a combinatorial attack on numerous oncogenic pathways. Hsp90 family consist of four isoforms; Hsp90α, Hsp90β, Grp94 and Trap-1. The development of Hsp90 isoform-selective inhibitors represent an alternative approach towards the treatment of cancer that may limit some of the detriments. We demonstrate novel Hsp90 inhibitors, on prostate and bladder cancer cells, which shows both potent antiproliferative effects and specific selectivity for Hsp90β. Methods: PC3MM2, LNCap-LN3, C4-2b, LAPC4 (prostate cancer) and T24, UC3 (bladder cancer) cancer cells were utilized. Cell Titer-Glo luminescent anti-proliferative assay was used to determine the IC50 numbers after 72h treatment. Trypan Blue Cytotoxicity assay was performed for 24h treatment with increasing concentrations of KUNB inhibitors. Effects of KUNB inhibitors on Hsp90’s client protein degradation were investigated by Western Blot. Results: KUNB31 manifested an IC50 of 3.00 µM against UC3 bladder cancer cells, UC3 cells were then evaluated via western blot analyses of known Hsp90α- and Hsp90β-dependent client proteins following treatment with KUNB31 for 24 hours. The data showed that, KUNB31 would not induce the heat shock response like 17AAG, and did cause Hsp90β related protein degradation (CXCR4). Moreover, Hsp27, PKM2, Her2, Hsf-1and Akt all showed degradation to different extent. KUNB105 exhibited potent anti-proliferative in both prostate and bladder cancer cells. IC50 number was determined as 1.24 µM for PC3MM2, 1.18 µM for LNCap-LN3, 1.03 µM for C4-2b, 2.56 µM for LAPC4, 0.20 µM for T24, and 0.30 µM for UC3 cancer cells. Conclusions: KUNB novel Hsp90β selective inhibitors, exhibit potent anti-proliferative and cytotoxic activity along with client protein degradation, without induction of HSR in prostate and bladder cancer cell lines. KUNB compound’s selective inhibition on Hsp90β isomers supports the development of Hsp90-selective inhibitors as a method to overcome the detriments associated with pan-inhibition in cancer treatment.

scholarly journals Polyporus polysaccharide regulates proliferation, apoptosis and migration of bladder cancer cells by polarizing macrophages to M1 subtype in tumor microenvironment

2020 ◽  
Author(s):  
Wenyu Jia ◽  
Siwan Luo ◽  
Gena Lai ◽  
Shiqi Li ◽  
Shuai Huo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bladder Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Activated Macrophages ◽  
Tnf Α ◽  
Bladder Cancer Cells ◽  
And Migration ◽  
Mesenchymal Transformation ◽  
M1 Type

Abstract BackgroundPolyporus polysaccharide (PPS), an active ingredient of traditional Chinese medicinal Polyporus umbellatus, has multiple biological functions, such as anti-cancer, immune-regulating and hepatoprotective activities. The purpose of this study was to investigate the mechanism of PPS activated macrophages in the treatment of bladder cancer.Methods100 ng/mL Phorbol myristate acetate (PMA) was used to induce THP-1 human leukemic cells as a macrophage model. Flow cytometry was used to detect the expression of CD14 and CD68 to verify the establishment of macrophage model. After that, Macrophages derived from THP-1 were treated with different concentrations of PPS (1,10 and 100 ug/mL). Flow cytometry and RT-PCR were used to detected the expression of CD16, CD23, CD86, CD40 and interleukin (IL)-Iβ, iNOS mRNA. ELISA was used to test the change of IL-1β and TNF-α in macrophage after the treatment with PPS. The conditioned medium from PPS-polarized macrophages was used to detect the effect of activated macrophages on bladder cancer. MTT assay, 5-ethynyl-2¢-deoxyuridine assay, flow cytometry, Transwell assay, and Western blot analysis were used to detect the effects of polarized macrophages on the viability, proliferation, apoptosis, and migration of bladder cancer cells. Western blot was also used to analysis the change of JAK2/NF-κB pathway protein.ResultsPPS promoted the expression of pro-inflammatory factors, such as IL-Iβ, TNF-α and iNOS, and surface molecules CD86, CD16, CD23, and CD40 in macrophages and then polarized macrophages to M1 type. The results demonstrated that activated macrophages inhibited the proliferation of bladder cancer cells, regulated their apoptosis, and inhibited migration and epithelial–mesenchymal transformation (EMT). JAK2/NF-κB pathways were downregulated in the anti-bladder cancer process of activated macrophages. ConclusionThe findings indicated that PPS inhibited the proliferation and progression of bladder cancer by the polarization of macrophages to M1 type, and JAK2/NF-κB pathway was downregulated in the process of anti-bladder cancer.

Antiproliferative effects of bacillus Calmette-Gu�rin and interferon ?2b on human bladder cancer cells in vitro

1995 ◽  
Vol 41 (5) ◽  
pp. 309-316 ◽  
Author(s):  
Kimberley Pryor ◽  
Phillip Stricker ◽  
Pamela Russell ◽  
David Golovsky ◽  
Ronald Penny
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Antiproliferative Effects ◽  
Bladder Cancer Cells ◽  
Human Bladder Cancer Cells

scholarly journals A positive feedback loop between TAZ and miR-942-3p modulates proliferation, migration, epithelial-mesenchymal transition process and glycometabolism in human bladder cancer

2020 ◽  
Author(s):  
Feifan Wang ◽  
Mengjing Fan ◽  
Xuejian Zhou ◽  
Yanlan Yu ◽  
Yueshu Cai ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Feedback Loop ◽  
Epithelial Mesenchymal Transition ◽  
Biological Functions ◽  
Positive Feedback Loop ◽  
Mesenchymal Transition ◽  
Qrt Pcr ◽  
Bladder Cancer Cells

Abstract Background: Transcriptional co-activator with PDZ-binding motif (TAZ) has been reported to involve in tumor progression, epithelial-mesenchymal transition (EMT) process and glycometabolism modulation. Herein, the underlying molecular mechanisms of TAZ-induced biological effects in bladder cancer were discovered; Methods: qRT-PCR, western blot and immunohistochemistry were performed to determine the level of TAZ in bladder cancer cells and tissues; CCK-8 assay, Colony formation assay, wound healing assay and Transwell assay were performed to evaluate the functions of TAZ, miR-942-3p and GAS1. qRT-PCR and western blot were used to determine the expression levels of related genes. Chromatin immunoprecipitation and dual-luciferase reporter assay confirmed the interaction between TAZ and miR-942. In vivo tumorigenesis assay and colorimetric assay of glycolysis were also conducted; Results: We determined the upregulation and vital roles of TAZ in bladder cancer. TAZ-induced upregulation of miR-942-3p amplified upstream signaling by inhibiting the expression of large tumor suppressor 2 (LATS2, a TAZ inhibitor). MiR-942-3p attenuated the suppression of cell proliferation, EMT process and glycolysis induced by TAZ knockdown. Further, miR-942-3p resulted in restrained expression of growth arrest-specific 1 (GAS1) to modulate biological functions; Conclusion: Our study identified a novel positive feedback loop between TAZ and miR-942-3p that regulates biological functions in bladder cancer cells via GAS1 expression, and illustrated that TAZ and miR-942-3p might be potential therapeutic targets for bladder cancer treatment.

scholarly journals Cheliensisin A (Chel A) induces apoptosis in human bladder cancer cells by promoting PHLPP2 protein degradation

Oncotarget ◽  
2016 ◽  
Vol 7 (41) ◽  
pp. 66689-66699 ◽  
Author(s):  
Ruowen Zhang ◽  
Xun Che ◽  
Jingjie Zhang ◽  
Yang Li ◽  
Jingxia Li ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Protein Degradation ◽  
Cancer Cells ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Bladder Cancer Cells ◽  
Human Bladder Cancer Cells

The Apoptosis Mechanism of Epirubicin Combined with BCG on Human Bladder Cancer Cells

2020 ◽  
Vol 20 (13) ◽  
pp. 1571-1581 ◽  
Author(s):  
Yang Luo ◽  
Xiaoyi Fu ◽  
Bin Han ◽  
Fafu Zhang ◽  
Lihong Yuan ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Caspase 3 ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Bladder Cancer Cells ◽  
Human Bladder Cancer Cells

Aims: The purpose of our study was to explore the combination effect of epirubicin and Bacillus Calmette Guerin (BCG) and its mechanism. Background: Bladder cancer is a threat to human health worldwide. Commonly used chemotherapy drugs and biotherapy have significant therapeutic effects on bladder cancer, but the mechanism and combined effects are still unclear. Objective: To evaluate the anti-cancer effect of epirubicin combined with BCG on human bladder cancer cells, our studies were carried out. Methods: The viability of human bladder cancer cells with epirubicin and/or BCG treatments was examined by Cell Counting Kit-8 (CCK-8) assay. Apoptosis and cell cycle phase were determined by flow cytometry analysis. Pre-apoptosis factors of caspase-3, p53, B-cell lymphoma 2 associated X protein (Bax) and anti-apoptosis factor of B-cell lymphoma 2 (Bcl2) were detected by western blot. Results: The viability of human bladder cancer with epirubicin or BCG treatment was decreased and the viability with epirubicin combined with BCG treatment was decreased more, which were determined by CCK-8 assay. Both epirubicin and BCG increased the apoptosis rate of human bladder cancer and arrested more cells into G0/G1 phase, which were tested by flow cytometry. The expression of caspase-3, p53 and Bax was increased and the expression of Bcl-2 was decreased with epirubicin treatment on human bladder cells, which were analyzed by western blot. The expression of caspase-3 and p53 was increased with BCG treatment, which was examined by western blot. Conclusion: Epirubicin induced apoptosis in human bladder cancer cells by up-regulating the expression of proapoptotic factors (caspase-3, p53 and Bax) and down-regulating the expression of anti-apoptotic factor (Bcl-2). BCG promoted apoptosis of human bladder cancer cells by up-regulating the expression of caspase-3 and p53. BCG plays a potential role at the time of the combination of epirubicin and BCG on bladder cancer cells in early stage. Both epirubicin and BCG affected cell cycle distribution via arresting more bladder cancer cells at G0/G1 phase, which ultimately led bladder cancer proliferation in vitro and promoted apoptosis.

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