We are analyzing https://link.springer.com/article/10.1186/s13046-014-0059-8.

Title:
Histone deacetylase 8 is deregulated in urothelial cancer but not a target for efficient treatment | Journal of Experimental & Clinical Cancer Research
Description:
Background Previous studies have shown that class-I histone deacetylase (HDAC) 8 mRNA is upregulated in urothelial cancer tissues and urothelial cancer cell lines compared to benign controls. Using urothelial cancer cell lines we evaluated whether specific targeting of HDAC8 might be a therapeutic option in bladder cancer treatment. Methods We conducted siRNA-mediated knockdown and specific pharmacological inhibition of HDAC8 with the three different inhibitors compound 2, compound 5, and compound 6 in several urothelial carcinoma cell lines with distinct HDAC8 expression profiles. Levels of HDAC and marker proteins were determined by western blot analysis and mRNA levels were measured by quantitative real-time PCR. Cellular effects of HDAC8 suppression were analyzed by ATP assay, flow cytometry, colony forming assay and migration assay. Results Efficient siRNA-mediated knockdown of HDAC8 reduced proliferation up to 45%. The HDAC8 specific inhibitors compound 5 and compound 6 significantly reduced viability of all urothelial cancer cell lines (IC50 9 – 21 μM). Flow cytometry revealed only a slight increase in the sub-G1 fraction indicating a limited induction of apoptosis. Expression of thymidylate synthase was partly reduced; PARP-cleavage was not detected. The influence of the pharmacological inhibition on clonogenic growth and migration show a cell line- and inhibitor-dependent reduction with the strongest effects after treatment with compound 5 and compound 6. Conclusions Deregulation of HDAC8 is frequent in urothelial cancer, but neither specific pharmacological inhibition nor siRNA-mediated knockdown of HDAC8 impaired viability of urothelial cancer cell lines in a therapeutic useful manner. Accordingly, HDAC8 on its own is not a promising drug target in bladder cancer.
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Keywords {🔍}

hdac, cell, cancer, cells, expression, pubmed, urothelial, lines, knockdown, figure, article, treatment, google, scholar, histone, compound, protein, cas, inhibition, effects, control, sirna, analysis, umuc, vmcub, germany, inhibitors, carcinoma, uccs, inhibitor, observed, deacetylase, significant, mrna, class, acetylation, pharmacological, authors, blot, αtubulin, size, western, proliferation, compared, bladder, growth, cycle, level, full, specific,

Topics {✒️}

hrp-conjugated goat-anti-mouse antibody hrp-conjugated goat-anti-rabbit antibody hrp-conjugated rabbit-anti-goat antibody 4-naphtyl-benzoil hydroxamic acid quantitative real-time pcr methotrexate/vinblastine/doxorubicin/cisplatin anti-α-tubulin b-512 eco1-dependent cohesin acetylation apparent g2/m-arrest icud-eau international consultation keratinocyte serum-free medium article download pdf pi3k/akt/mtor signaling t-cell derived lymphoma g0/g1 arrest induced cell line-dependent manner conducted sirna-mediated knockdown isoenzyme-selective hdac inhibitor open access license invasive bladder cancer quantitative rt-pcr analysis long-term survival results metastatic bladder cancer cytoplasmic protein α-tubulin pan hdac-inhibitor saha cancer epigenetics bladder cancer treatment loading control α-tubulin hdac8-sirna mediated knockdown sirna-mediated hdac8 knockdown decreased s-phase fraction celltiter-glo reagent real-time pcr cell cycle arrest high hdac activity cell lines vm-cub1 full size image normal uroepithelial cultures normal uroepithelial control authors’ original file privacy choices/manage cookies p53 c-terminal domain muscle invasive carcinoma g2/m-arrest normal uroepithelial cells urothelial carcinoma-neoadjuvant um-uc-3 cells compared urothelial cancer tissues urothelial cancer cells g0/g1 arrest

Questions {❓}

Witt O, Deubzer HE, Milde T, Oehme I: HDAC family: What are the cancer relevant targets?

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         description:Previous studies have shown that class-I histone deacetylase (HDAC) 8 mRNA is upregulated in urothelial cancer tissues and urothelial cancer cell lines compared to benign controls. Using urothelial cancer cell lines we evaluated whether specific targeting of HDAC8 might be a therapeutic option in bladder cancer treatment. We conducted siRNA-mediated knockdown and specific pharmacological inhibition of HDAC8 with the three different inhibitors compound 2, compound 5, and compound 6 in several urothelial carcinoma cell lines with distinct HDAC8 expression profiles. Levels of HDAC and marker proteins were determined by western blot analysis and mRNA levels were measured by quantitative real-time PCR. Cellular effects of HDAC8 suppression were analyzed by ATP assay, flow cytometry, colony forming assay and migration assay. Efficient siRNA-mediated knockdown of HDAC8 reduced proliferation up to 45%. The HDAC8 specific inhibitors compound 5 and compound 6 significantly reduced viability of all urothelial cancer cell lines (IC50 9 – 21 μM). Flow cytometry revealed only a slight increase in the sub-G1 fraction indicating a limited induction of apoptosis. Expression of thymidylate synthase was partly reduced; PARP-cleavage was not detected. The influence of the pharmacological inhibition on clonogenic growth and migration show a cell line- and inhibitor-dependent reduction with the strongest effects after treatment with compound 5 and compound 6. Deregulation of HDAC8 is frequent in urothelial cancer, but neither specific pharmacological inhibition nor siRNA-mediated knockdown of HDAC8 impaired viability of urothelial cancer cell lines in a therapeutic useful manner. Accordingly, HDAC8 on its own is not a promising drug target in bladder cancer.
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      headline:Histone deacetylase 8 is deregulated in urothelial cancer but not a target for efficient treatment
      description:Previous studies have shown that class-I histone deacetylase (HDAC) 8 mRNA is upregulated in urothelial cancer tissues and urothelial cancer cell lines compared to benign controls. Using urothelial cancer cell lines we evaluated whether specific targeting of HDAC8 might be a therapeutic option in bladder cancer treatment. We conducted siRNA-mediated knockdown and specific pharmacological inhibition of HDAC8 with the three different inhibitors compound 2, compound 5, and compound 6 in several urothelial carcinoma cell lines with distinct HDAC8 expression profiles. Levels of HDAC and marker proteins were determined by western blot analysis and mRNA levels were measured by quantitative real-time PCR. Cellular effects of HDAC8 suppression were analyzed by ATP assay, flow cytometry, colony forming assay and migration assay. Efficient siRNA-mediated knockdown of HDAC8 reduced proliferation up to 45%. The HDAC8 specific inhibitors compound 5 and compound 6 significantly reduced viability of all urothelial cancer cell lines (IC50 9 – 21 μM). Flow cytometry revealed only a slight increase in the sub-G1 fraction indicating a limited induction of apoptosis. Expression of thymidylate synthase was partly reduced; PARP-cleavage was not detected. The influence of the pharmacological inhibition on clonogenic growth and migration show a cell line- and inhibitor-dependent reduction with the strongest effects after treatment with compound 5 and compound 6. Deregulation of HDAC8 is frequent in urothelial cancer, but neither specific pharmacological inhibition nor siRNA-mediated knockdown of HDAC8 impaired viability of urothelial cancer cell lines in a therapeutic useful manner. Accordingly, HDAC8 on its own is not a promising drug target in bladder cancer.
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         Cell cycle arrest
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         Immunology
         Apoptosis
         Oncology
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      name:Department of Urology, Heinrich-Heine-University, Medical Faculty, Duesseldorf, Germany
      name:Department of Urology, Heinrich-Heine-University, Medical Faculty, Duesseldorf, Germany

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