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Victor E. Marquez - One of the best experts on this subject based on the ideXlab platform.
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Enhanced growth inhibition by combined DNA methylation/HDAC inhibitors in lung tumor cells with silenced CDKN2A.
International journal of oncology, 2010Co-Authors: Min Chen, Victor E. Marquez, Frederic J. Kaye, Donna Voeller, Giuseppe Giaccone, Patricia S. Steeg, Maria Zajac-kayeAbstract:Aberrant hypermethylation at CpG sites within the CDKN2A gene is associated with silencing and has been proposed as a target for reactivation using both DNA methylation and histone deacetylation inhibitors. This study investigates the role of selecting tumor samples with a silenced as compared to deleted CDKN2A locus when assessing the efficacy of DNA methyltransferase inhibitor, Zebularine, combined with the HDAC inhibitor, depsipeptide. Non-small cell lung cancer cell lines with defined CDKN2A status were analyzed by MTS assay to determine the effect of Zebularine or Zebularine combined with depsipeptide on tumor cell growth. We observed that Zebularine treatment resulted in inhibition of cell growth in 11 out of 12 lung cancer cell lines with silenced CDKN2A, but no cell growth inhibition was detected in the 7 lung cancer cell lines tested with deleted CDKN2A (p>0.001). In addition, we found that the combi- nation of 30 μM Zebularine and 6 or 7 nM depsipeptide resulted in a synergistic inhibition of cell growth in tumor cells with silenced CDKN2A (p
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DNA (Cytosine-C5) methyltransferase inhibition by oligodeoxyribonucleotides containing 2-(1H)-pyrimidinone (Zebularine aglycon) at the enzymatic target site
Biochemical pharmacology, 2009Co-Authors: Dana M. Van Bemmel, Victor E. Marquez, Ramon Eritja, Adam S. Brank, Judith K. ChristmanAbstract:Aberrant cytosine methylation in promoter regions leads to gene silencing associated with cancer progression. A number of DNA methyltransferase inhibitors are known to reactivate silenced genes; including 5-azacytidine and 2-(1H)-pyrimidinone riboside (Zebularine). Zebularine is a more stable, less cytotoxic inhibitor compared to 5-azacytidine. To determine the mechanistic basis for this difference, we carried out a detailed comparisons of the interaction between purified DNA methyltransferases and oligodeoxyribonucleotides (ODNs) containing either 5-azacytosine or 2-(1H)-pyrimidinone in place of the cytosine targeted for methylation. When incorporated into small ODNs, the rate of C5 DNA methyltransferase inhibition by both nucleosides is essentially identical. However, the stability and reversibility of the enzyme complex in the absence and presence of cofactor differs. 5-Azacytosine ODNs form complexes with C5 DNA methyltransferases that are irreversible when the 5-azacytosine ring is intact. ODNs containing 2-(1H)-pyrimidinone at the enzymatic target site are competitive inhibitors of both prokaryotic and mammalian DNA C5 methyltransferases. We determined that the ternary complexes between the enzymes, 2-(1H)-pyrimidinone inhibitor, and the cofactor S-adenosyl methionine are maintained through the formation of a reversible covalent interaction. The differing stability and reversibility of the covalent bonds may partially account for the observed differences in cytotoxicity between Zebularine and 5-azacytidine inhibitors.
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Activation of p16 gene silenced by DNA methylation in cancer cells by phosphoramidate derivatives of 2’-deoxyZebularine
Journal of medicinal chemistry, 2008Co-Authors: Christine B. Yoo, Peter A Jones, Rocco Valente, Costantino Congiatu, Federica Gavazza, Annette Angel, Maqbool A. Siddiqui, Christopher Mcguigan, Victor E. MarquezAbstract:We report herein the application of the phosphoramidate ProTide technology to improve the metabolism of the DNA methytransferase inhibitor, Zebularine (Z). Zebularine is a riboside that must undergo a complex metabolic transformation before reaching the critical 2'-deoxyZebularine 5'-triphosphate (dZTP). Because 2'-deoxyZebularine (dZ) is not phosphorylated and therefore inactive, the ProTide strategy was employed to bypass the lack of phosphorylation of dZ and the inefficient reduction of Zebularine 5'-diphosphate by ribonucleotide-diphosphate reductase required for Zebularine. Several compounds were identified as more potent inhibitors of DNA methylation and stronger inducers of p16 tumor suppressor gene than Zebularine. However, their activity was dependent on the administration of thymidine to overcome the potent inhibition of thymidylate synthase (TS) and deoxycytidine monophosphate (dCMP) deaminase by dZMP, which deprives cells of essential levels of thymidine. Intriguingly, the activity of the ProTides was cell line-dependent, and activation of p16 was manifest only in Cf-Pac-1 pancreatic ductal adenocarcinoma cells.
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Induction of apoptosis in MMTV/PyMT mammary gland tumors with oral Zebularine
Cancer Research, 2008Co-Authors: Min Chen, Victor E. Marquez, Melinda G. Hollingshead, Sherry X. Yang, Kent W. Hunter, Donna Voeller, Giuseppe Giaccone, Patricia S. Steeg, Maria Zajac-kayeAbstract:2611 Zebularine, a synthetic analog of cytidine and a potent inhibitor of cytidine deaminase, has recently been identified as a DNA methylation inhibitor. In addition, nude mice with EJ6 bladder cancer xenografts showed induction of p16 expression and reduction of tumor growth following oral Zebularine as compared to untreated controls (Cheng et al JNCI 95 p399 2003). We studied the effect of Zebularine on mammary tumor growth in MMTV-PyMT transgenic mice which develop mammary tumors within 60 days with 100 percent penetrance. The mice were randomized at 46 days of age into control (n=25) and Zebularine treated (n=25) groups, which were given 5 mg/ml of Zebularine in drinking water. Tumors were measured twice weekly and the median tumor weight was calculated for each group. We found a significant delay in the occurrence of tumors in both the axillary and inguinal mammary glands in Zebularine treated mice as compared to untreated controls. We observed a statistically significant (p = 0.01) reduction in total tumor burden at 94 days of age when the mice were sacrificed. To determine the mechanism of the Zebularine tumor effect we examined the histology of the treated and control tumors. After 48 days of drug treatment the tumors were predominantly necrotic compared to the untreated animals and a high apoptotic index was observed as early as 13 days by tunel assay. A negligible level of apoptotic cells was found in tumors from control animals. Although we did not observe induction of p16 levels after 13, 23 and 48 days of Zebularine treatment, we did observe induction of p21 expression, suggesting a tissue specific effect on patterns of gene demethylation. Immunoblot analysis showed depletion of DNMT1 and partial depletion of DNMT3b after Zebularine treatment. To investigate the effect of Zebularine on global gene expression profiles, microarray analyses were performed on tumors that were excised at 13, 23 and 48 days from both treated and untreated mice. This analysis confirmed p21 induction and the absence of p16 induction. In addition, we identified upregulation of four additional methylation regulated genes (1.5 to 2.4 fold increase) as well as a set of candidate cancer genes that may play a role in cell growth and apoptosis. Our results suggest that the pattern of DNA demethylation following Zebularine treatment is restricted to specific sets of genes and that cell death and necrosis contribute to the delay in tumor growth observed in mice continuously exposed to the oral intake of the drug.
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induction of apoptosis in mmtv pymt mammary gland tumors with oral Zebularine
Cancer Research, 2008Co-Authors: Min Chen, Victor E. Marquez, Melinda G. Hollingshead, Sherry X. Yang, Kent W. Hunter, Donna Voeller, Giuseppe Giaccone, Patricia S. Steeg, Maria ZajackayeAbstract:2611 Zebularine, a synthetic analog of cytidine and a potent inhibitor of cytidine deaminase, has recently been identified as a DNA methylation inhibitor. In addition, nude mice with EJ6 bladder cancer xenografts showed induction of p16 expression and reduction of tumor growth following oral Zebularine as compared to untreated controls (Cheng et al JNCI 95 p399 2003). We studied the effect of Zebularine on mammary tumor growth in MMTV-PyMT transgenic mice which develop mammary tumors within 60 days with 100 percent penetrance. The mice were randomized at 46 days of age into control (n=25) and Zebularine treated (n=25) groups, which were given 5 mg/ml of Zebularine in drinking water. Tumors were measured twice weekly and the median tumor weight was calculated for each group. We found a significant delay in the occurrence of tumors in both the axillary and inguinal mammary glands in Zebularine treated mice as compared to untreated controls. We observed a statistically significant (p = 0.01) reduction in total tumor burden at 94 days of age when the mice were sacrificed. To determine the mechanism of the Zebularine tumor effect we examined the histology of the treated and control tumors. After 48 days of drug treatment the tumors were predominantly necrotic compared to the untreated animals and a high apoptotic index was observed as early as 13 days by tunel assay. A negligible level of apoptotic cells was found in tumors from control animals. Although we did not observe induction of p16 levels after 13, 23 and 48 days of Zebularine treatment, we did observe induction of p21 expression, suggesting a tissue specific effect on patterns of gene demethylation. Immunoblot analysis showed depletion of DNMT1 and partial depletion of DNMT3b after Zebularine treatment. To investigate the effect of Zebularine on global gene expression profiles, microarray analyses were performed on tumors that were excised at 13, 23 and 48 days from both treated and untreated mice. This analysis confirmed p21 induction and the absence of p16 induction. In addition, we identified upregulation of four additional methylation regulated genes (1.5 to 2.4 fold increase) as well as a set of candidate cancer genes that may play a role in cell growth and apoptosis. Our results suggest that the pattern of DNA demethylation following Zebularine treatment is restricted to specific sets of genes and that cell death and necrosis contribute to the delay in tumor growth observed in mice continuously exposed to the oral intake of the drug.
Fraidoon Kavoosi - One of the best experts on this subject based on the ideXlab platform.
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Effect of Zebularine in Comparison to and in Combination with Trichostatin A on CIP/KIP Family (p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2), DNMTs (DNMT1, DNMT3a, and DNMT3b), Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) Gene Express
Asian Pacific journal of cancer prevention : APJCP, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background A pattern of epigenetic modifications and changes, DNA methylation and histone modification, is central to many human cancers. A variety of tumor suppressor genes (TSGs) have been demonstrated to be silenced because of histone deacetylation and DNA hypermethylation in several cancers. Recent in vitro studies have shown that two known mechanisms of epigenetic alteration consisting of methylation and histone deacetylation seem to be the best candidate mechanisms for inactivation of CIP/KIP family (p21Cip1/Waf1/Sdi1, and p27Kip1) in numerous cancers. Numerous investigations have indicated that DNA demethylating and histone deacetylase inhibitors (HDACIs) can restore the CIP/KIP family gene expression. Previously, we evaluated the effect of trichostatin A (TSA) and 5-aza-2'-deoxycytidine (5-AZA-CdR) on hepatocellular carcinoma (HCC). The present study was designed to investigate the effect of Zebularine in comparison to and in combination with trichostatin A on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNMT1, DNMT3a and DNMT3b, Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) gene expression, cell growth inhibition and apoptosis induction in colon cancer LS 174T cell line. Materials and methods The colon cancer LS 174T cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and trichostatin A can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity.
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effect of Zebularine in comparison to and in combination with trichostatin a on cip kip family p21cip1 waf1 sdi1 p27kip1 and p57kip2 dnmts dnmt1 dnmt3a and dnmt3b class i hdacs hdacs 1 2 3 and class ii hdacs hdacs 4 5 6 gene expression cell growth in
Asian Pacific Journal of Cancer Prevention, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background A pattern of epigenetic modifications and changes, DNA methylation and histone modification, is central to many human cancers. A variety of tumor suppressor genes (TSGs) have been demonstrated to be silenced because of histone deacetylation and DNA hypermethylation in several cancers. Recent in vitro studies have shown that two known mechanisms of epigenetic alteration consisting of methylation and histone deacetylation seem to be the best candidate mechanisms for inactivation of CIP/KIP family (p21Cip1/Waf1/Sdi1, and p27Kip1) in numerous cancers. Numerous investigations have indicated that DNA demethylating and histone deacetylase inhibitors (HDACIs) can restore the CIP/KIP family gene expression. Previously, we evaluated the effect of trichostatin A (TSA) and 5-aza-2'-deoxycytidine (5-AZA-CdR) on hepatocellular carcinoma (HCC). The present study was designed to investigate the effect of Zebularine in comparison to and in combination with trichostatin A on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNMT1, DNMT3a and DNMT3b, Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) gene expression, cell growth inhibition and apoptosis induction in colon cancer LS 174T cell line. Materials and methods The colon cancer LS 174T cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and trichostatin A can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity.
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investigation of the effect of Zebularine in comparison to and in combination with trichostatin a on p21cip1 waf1 sdi1 p27kip1 p57kip2 dna methyltransferases and histone deacetylases in colon cancer ls 180 cell line
Asian Pacific Journal of Cancer Prevention, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background The heart of the cell cycle regulatory machine is a group of enzymes named cyclin-dependent kinases (Cdks). The active form of these enzymes includes a kinase and its partner, a cyclin. The regulation of cyclin-Cdk complexes is provided by Cdk inhibitors (CKIs) such as Cip/Kip family comprising p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2. The hypermethylation and deacetylation of Cip/Kip gene family seem to be frequent in numerous cancers. It has been indicated that increased expression of DNMTs and HDACs contributes to cancer induction. Previously, we reported the effect of DNA demethylating agents and histone deacetylase inhibitors on histone deacetylase 1, DNA methyltransferase 1, and CIP/KIP family in colon cancer. The current study was designed to evaluate the effect of Zebularine in comparison to and in combination with trichostatin A (TSA) on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNA methyltransferases (DNMT1, 3a and 3b) and histone deacetylases (HDAC1, 2, and 3) genes expression, cell growth inhibition and apoptosis induction in colon cancer LS 180 cell line. Materials and methods The colon cancer LS 180 cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and TSA can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity. .
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Investigation of the Effect of Zebularine in Comparison to and in Combination with Trichostatin A on p21Cip1/Waf1/ Sdi1, p27Kip1, p57Kip2, DNA Methyltransferases and Histone Deacetylases in Colon Cancer LS 180 Cell Line
Asian Pacific journal of cancer prevention : APJCP, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background The heart of the cell cycle regulatory machine is a group of enzymes named cyclin-dependent kinases (Cdks). The active form of these enzymes includes a kinase and its partner, a cyclin. The regulation of cyclin-Cdk complexes is provided by Cdk inhibitors (CKIs) such as Cip/Kip family comprising p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2. The hypermethylation and deacetylation of Cip/Kip gene family seem to be frequent in numerous cancers. It has been indicated that increased expression of DNMTs and HDACs contributes to cancer induction. Previously, we reported the effect of DNA demethylating agents and histone deacetylase inhibitors on histone deacetylase 1, DNA methyltransferase 1, and CIP/KIP family in colon cancer. The current study was designed to evaluate the effect of Zebularine in comparison to and in combination with trichostatin A (TSA) on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNA methyltransferases (DNMT1, 3a and 3b) and histone deacetylases (HDAC1, 2, and 3) genes expression, cell growth inhibition and apoptosis induction in colon cancer LS 180 cell line. Materials and methods The colon cancer LS 180 cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and TSA can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity. .
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Effect of valproic acid and Zebularine on SOCS-1 and SOCS-3 gene expression in colon carcinoma SW48 cell line.
Experimental oncology, 2020Co-Authors: Masumeh Sanaei, Fraidoon Kavoosi, H BehjooAbstract:BACKGROUND Two epigenetic modifications such as histone acetylation and DNA methylation have been known as critical players of gene regulation. Hypermethylation and deacetylation of suppressors of cytokine signaling family SOCS-1 and SOCS-3 have been shown in many solid cancers. Previously, we evaluated the effect of 5-aza-2'-deoxycytidine and valproic acid on hepatocellular carcinoma and colon cancer cells. AIM The present study was designed to assess the effect of valproic acid in comparison to Zebularine on SOCS-1 and SOCS-3 gene expression, cell growth inhibition and apoptosis induction in colon carcinoma SW48 cell line. MATERIALS AND METHODS SW48 cells were treated with valproic acid or Zebularine for 24 h and 48 h. The effect of the compounds on cell viability, SOCS-1 and SOCS-3 gene expression, and apoptosis induction was evaluated. Reverse transcription polymerase chain reaction analysis and flow cytometry were applied. RESULTS Both agents inhibited cell growth in a time- and dose-dependent fashion. The apoptotic effect was observed in cells treated with valproic acid (7.5 μM) but not Zebularine (75 μM). The valproic acid but not Zebularine upregulated SOCS-1 and SOCS-3 gene expression. CONCLUSION Epigenetic modulation can reactivate silenced tumor suppressor genes SOCS-1 and SOCS-3 through histone acetylation resulting in apoptosis induction.
Masumeh Sanaei - One of the best experts on this subject based on the ideXlab platform.
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Effect of Zebularine in Comparison to and in Combination with Trichostatin A on CIP/KIP Family (p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2), DNMTs (DNMT1, DNMT3a, and DNMT3b), Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) Gene Express
Asian Pacific journal of cancer prevention : APJCP, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background A pattern of epigenetic modifications and changes, DNA methylation and histone modification, is central to many human cancers. A variety of tumor suppressor genes (TSGs) have been demonstrated to be silenced because of histone deacetylation and DNA hypermethylation in several cancers. Recent in vitro studies have shown that two known mechanisms of epigenetic alteration consisting of methylation and histone deacetylation seem to be the best candidate mechanisms for inactivation of CIP/KIP family (p21Cip1/Waf1/Sdi1, and p27Kip1) in numerous cancers. Numerous investigations have indicated that DNA demethylating and histone deacetylase inhibitors (HDACIs) can restore the CIP/KIP family gene expression. Previously, we evaluated the effect of trichostatin A (TSA) and 5-aza-2'-deoxycytidine (5-AZA-CdR) on hepatocellular carcinoma (HCC). The present study was designed to investigate the effect of Zebularine in comparison to and in combination with trichostatin A on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNMT1, DNMT3a and DNMT3b, Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) gene expression, cell growth inhibition and apoptosis induction in colon cancer LS 174T cell line. Materials and methods The colon cancer LS 174T cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and trichostatin A can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity.
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effect of Zebularine in comparison to and in combination with trichostatin a on cip kip family p21cip1 waf1 sdi1 p27kip1 and p57kip2 dnmts dnmt1 dnmt3a and dnmt3b class i hdacs hdacs 1 2 3 and class ii hdacs hdacs 4 5 6 gene expression cell growth in
Asian Pacific Journal of Cancer Prevention, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background A pattern of epigenetic modifications and changes, DNA methylation and histone modification, is central to many human cancers. A variety of tumor suppressor genes (TSGs) have been demonstrated to be silenced because of histone deacetylation and DNA hypermethylation in several cancers. Recent in vitro studies have shown that two known mechanisms of epigenetic alteration consisting of methylation and histone deacetylation seem to be the best candidate mechanisms for inactivation of CIP/KIP family (p21Cip1/Waf1/Sdi1, and p27Kip1) in numerous cancers. Numerous investigations have indicated that DNA demethylating and histone deacetylase inhibitors (HDACIs) can restore the CIP/KIP family gene expression. Previously, we evaluated the effect of trichostatin A (TSA) and 5-aza-2'-deoxycytidine (5-AZA-CdR) on hepatocellular carcinoma (HCC). The present study was designed to investigate the effect of Zebularine in comparison to and in combination with trichostatin A on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNMT1, DNMT3a and DNMT3b, Class I HDACs (HDACs 1, 2, 3) and Class II HDACs (HDACs 4, 5, 6) gene expression, cell growth inhibition and apoptosis induction in colon cancer LS 174T cell line. Materials and methods The colon cancer LS 174T cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and trichostatin A can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity.
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investigation of the effect of Zebularine in comparison to and in combination with trichostatin a on p21cip1 waf1 sdi1 p27kip1 p57kip2 dna methyltransferases and histone deacetylases in colon cancer ls 180 cell line
Asian Pacific Journal of Cancer Prevention, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background The heart of the cell cycle regulatory machine is a group of enzymes named cyclin-dependent kinases (Cdks). The active form of these enzymes includes a kinase and its partner, a cyclin. The regulation of cyclin-Cdk complexes is provided by Cdk inhibitors (CKIs) such as Cip/Kip family comprising p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2. The hypermethylation and deacetylation of Cip/Kip gene family seem to be frequent in numerous cancers. It has been indicated that increased expression of DNMTs and HDACs contributes to cancer induction. Previously, we reported the effect of DNA demethylating agents and histone deacetylase inhibitors on histone deacetylase 1, DNA methyltransferase 1, and CIP/KIP family in colon cancer. The current study was designed to evaluate the effect of Zebularine in comparison to and in combination with trichostatin A (TSA) on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNA methyltransferases (DNMT1, 3a and 3b) and histone deacetylases (HDAC1, 2, and 3) genes expression, cell growth inhibition and apoptosis induction in colon cancer LS 180 cell line. Materials and methods The colon cancer LS 180 cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and TSA can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity. .
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Investigation of the Effect of Zebularine in Comparison to and in Combination with Trichostatin A on p21Cip1/Waf1/ Sdi1, p27Kip1, p57Kip2, DNA Methyltransferases and Histone Deacetylases in Colon Cancer LS 180 Cell Line
Asian Pacific journal of cancer prevention : APJCP, 2020Co-Authors: Masumeh Sanaei, Fraidoon KavoosiAbstract:Background The heart of the cell cycle regulatory machine is a group of enzymes named cyclin-dependent kinases (Cdks). The active form of these enzymes includes a kinase and its partner, a cyclin. The regulation of cyclin-Cdk complexes is provided by Cdk inhibitors (CKIs) such as Cip/Kip family comprising p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2. The hypermethylation and deacetylation of Cip/Kip gene family seem to be frequent in numerous cancers. It has been indicated that increased expression of DNMTs and HDACs contributes to cancer induction. Previously, we reported the effect of DNA demethylating agents and histone deacetylase inhibitors on histone deacetylase 1, DNA methyltransferase 1, and CIP/KIP family in colon cancer. The current study was designed to evaluate the effect of Zebularine in comparison to and in combination with trichostatin A (TSA) on p21Cip1/Waf1/Sdi1, p27Kip1, p57Kip2, DNA methyltransferases (DNMT1, 3a and 3b) and histone deacetylases (HDAC1, 2, and 3) genes expression, cell growth inhibition and apoptosis induction in colon cancer LS 180 cell line. Materials and methods The colon cancer LS 180 cell line was cultured and treated with Zebularine and TSA. To determine cell viability, apoptosis, and the relative expression level of the genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. Results Both compounds significantly inhibited cell growth, and induced apoptosis. Furthermore, both compounds increased p21Cip1/Waf1/Sdi1, p27Kip1, and p57Kip2 significantly. Additionally, Zebularine and TSA decreased DNMTs and HDACs gene expression respectively. Conclusion The Zebularine and TSA can reactivate the CIP/KIP family through inhibition of DNMTs and HDACs genes activity. .
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Effect of valproic acid and Zebularine on SOCS-1 and SOCS-3 gene expression in colon carcinoma SW48 cell line.
Experimental oncology, 2020Co-Authors: Masumeh Sanaei, Fraidoon Kavoosi, H BehjooAbstract:BACKGROUND Two epigenetic modifications such as histone acetylation and DNA methylation have been known as critical players of gene regulation. Hypermethylation and deacetylation of suppressors of cytokine signaling family SOCS-1 and SOCS-3 have been shown in many solid cancers. Previously, we evaluated the effect of 5-aza-2'-deoxycytidine and valproic acid on hepatocellular carcinoma and colon cancer cells. AIM The present study was designed to assess the effect of valproic acid in comparison to Zebularine on SOCS-1 and SOCS-3 gene expression, cell growth inhibition and apoptosis induction in colon carcinoma SW48 cell line. MATERIALS AND METHODS SW48 cells were treated with valproic acid or Zebularine for 24 h and 48 h. The effect of the compounds on cell viability, SOCS-1 and SOCS-3 gene expression, and apoptosis induction was evaluated. Reverse transcription polymerase chain reaction analysis and flow cytometry were applied. RESULTS Both agents inhibited cell growth in a time- and dose-dependent fashion. The apoptotic effect was observed in cells treated with valproic acid (7.5 μM) but not Zebularine (75 μM). The valproic acid but not Zebularine upregulated SOCS-1 and SOCS-3 gene expression. CONCLUSION Epigenetic modulation can reactivate silenced tumor suppressor genes SOCS-1 and SOCS-3 through histone acetylation resulting in apoptosis induction.
Peter A Jones - One of the best experts on this subject based on the ideXlab platform.
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Activation of p16 gene silenced by DNA methylation in cancer cells by phosphoramidate derivatives of 2’-deoxyZebularine
Journal of medicinal chemistry, 2008Co-Authors: Christine B. Yoo, Peter A Jones, Rocco Valente, Costantino Congiatu, Federica Gavazza, Annette Angel, Maqbool A. Siddiqui, Christopher Mcguigan, Victor E. MarquezAbstract:We report herein the application of the phosphoramidate ProTide technology to improve the metabolism of the DNA methytransferase inhibitor, Zebularine (Z). Zebularine is a riboside that must undergo a complex metabolic transformation before reaching the critical 2'-deoxyZebularine 5'-triphosphate (dZTP). Because 2'-deoxyZebularine (dZ) is not phosphorylated and therefore inactive, the ProTide strategy was employed to bypass the lack of phosphorylation of dZ and the inefficient reduction of Zebularine 5'-diphosphate by ribonucleotide-diphosphate reductase required for Zebularine. Several compounds were identified as more potent inhibitors of DNA methylation and stronger inducers of p16 tumor suppressor gene than Zebularine. However, their activity was dependent on the administration of thymidine to overcome the potent inhibition of thymidylate synthase (TS) and deoxycytidine monophosphate (dCMP) deaminase by dZMP, which deprives cells of essential levels of thymidine. Intriguingly, the activity of the ProTides was cell line-dependent, and activation of p16 was manifest only in Cf-Pac-1 pancreatic ductal adenocarcinoma cells.
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Long-term epigenetic therapy with oral Zebularine has minimal side effects and prevents intestinal tumors in mice.
Cancer prevention research (Philadelphia Pa.), 2008Co-Authors: Christine B. Yoo, Victor E. Marquez, Allen S. Yang, Jody C. Chuang, Hyang-min Byun, Gerda Egger, Louis Dubeau, Tiffany I. Long, Peter W. Laird, Peter A JonesAbstract:Recent successes in the application of epigenetic drugs for the treatment of myelodysplastic syndrome have raised questions on the safety of long-term administration of DNA methylation inhibitors. We treated preweaned cancer prone ApcMin/+ (Min) mice continuously with the DNA methylation inhibitor Zebularine in their drinking water to determine the effects of the drug on normal mouse development as well as cancer prevention. Zebularine caused a tissue-specific reduction in DNA methylation at B1 short interspersed nucleotide elements in the small and large intestines of female Min mice but not in other organs examined after chronic oral treatment. No significant difference in the average weights of mice was observed during the treatment. In addition, analysis of global gene expression of colonic epithelial cells from the females indicated that only 3% to 6% of the genes were affected in their expression. We did not detect toxicity and abnormalities from the histopathologic analysis of liver and intestinal tissues. Lastly, we tested whether prevention of tumorigenesis can be achieved with chronic oral administration of Zebularine in Min mice. The average number of polyps in Min females decreased from 58 to 1, whereas the average polyp number remained unaffected in Min males possibly due to differential activity of aldehyde oxidase. Taken together, our results show for the first time that long-term oral administration of Zebularine causes a gender-specific abrogation of intestinal tumors while causing a tissue-specific DNA demethylation. Importantly, prolonged treatment of mice with epigenetic drugs resulted in only minor developmental and histologic changes.
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Zebularine: A Unique Molecule for an Epigenetically Based Strategy in Cancer Chemotherapy
Annals of the New York Academy of Sciences, 2005Co-Authors: Victor E. Marquez, James A. Kelley, Jonathan C Cheng, Christine B. Yoo, Riad Agbaria, Tisipi Ben‐kasus, Peter A JonesAbstract:1-(Beta-d-ribofuranosyl)-1,2-dihydropyrimidin-2-one (Zebularine) corresponds structurally to cytidine minus the exocyclic 4-amino group. The increased electrophilic character of its simple aglycon endows the molecule with unique biologic properties as a potent inhibitor of both cytidine deaminase and DNA cytosine methyltransferase. The latter activity makes Zebularine a promising antitumor agent that is hydrolytically stable, preferentially targets cancer cells, and shows activity both in vitro and in experimental animals, even after oral administration.
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Zebularine: a unique molecule for an epigenetically based strategy in cancer chemotherapy. The magic of its chemistry and biology.
Nucleosides nucleotides & nucleic acids, 2005Co-Authors: Victor E. Marquez, James A. Kelley, Joseph J. Barchi, Jonathan C Cheng, Christine B. Yoo, Kambhampati V. R. Rao, Riad Agbaria, Tsipi Ben-kasus, Peter A JonesAbstract:1-(beta-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one (Zebularine) is structurally 4-deamino cytidine. The increased electrophilic character of this simple aglycon endows the molecule with unique chemical and biological properties, making Zebularine a versatile starting material for the synthesis of complex nucleosides and an effective inhibitor of cytidine deaminase and DNA cytosine methyltransferase. Zebularine is a stable, antitumor agent that preferentially targets cancer cells and shows activity both in vitro and in experimental animals, even after oral administration.
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Zebularine: a new drug for epigenetic therapy.
Biochemical Society Transactions, 2004Co-Authors: C.b. Yoo, J.c. Cheng, Peter A JonesAbstract:Regulatory genes are often hypermethylated at their promoter 5' regions and silenced in cancer. Epigenetic therapy with DNA methylation inhibitors have been shown to result in the demethylation and reactivation of these genes. Zebularine is a recently discovered mechanism-based inhibitor of DNA methylation, and has received much attention for its potential in clinical use. Further studies exploring the effectiveness of Zebularine in a variety of settings could allow the development of novel therapies for cancer.
Elvar Theodorsson - One of the best experts on this subject based on the ideXlab platform.
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dna methylation inhibitor Zebularine confers stroke protection in ischemic rats
Translational Stroke Research, 2015Co-Authors: Hua Dock, Annette Theodorsson, Elvar TheodorssonAbstract:5-Aza-deoxycytidine (5-aza-dC) confers neuroprotection in ischemic mice by inhibiting DNA methylation. Zebularine is another DNA methylation inhibitor, less toxic and more stable in aqueous solutions and, therefore more biologically suitable. We investigated Zebularine’s effects on brain ischemia in a rat middle cerebral artery occlusion (MCAo) model in order to elucidate its therapeutic potential. Male Wistar wild-type (WT) rats were randomly allocated to three treatment groups, vehicle, Zebularine 100 μg, and Zebularine 500 μg. Saline (10 μL) or Zebularine (10 μL) was administered intracerebroventricularly 20 min before 45-min occlusion of the middle cerebral artery. Reperfusion was allowed after 45-min occlusion, and the rats were sacrificed at 24-h reperfusion. The brains were removed, sliced, and stained with 2 % 2,3,5-triphenyltetrazolium chloride (TTC) before measuring infarct size. Zebularine (500 μg) reduced infarct volumes significantly (p < 0.05) by 61 % from 20.7 ± 4.2 % in the vehicle treated to 8.1 ± 1.6 % in the Zebularine treated. Zebularine (100 μg) also reduced infarct volumes dramatically by 55 to 9.4 ± 1.2 %. The mechanisms behind this neuroprotection is not yet known, but the results agree with previous studies and support the notion that Zebularine-induced inhibition of DNA methyltransferase ameliorates ischemic brain injury in rats.
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DNA Methylation Inhibitor Zebularine Confers Stroke Protection in Ischemic Rats
Translational stroke research, 2015Co-Authors: Hua Dock, Annette Theodorsson, Elvar TheodorssonAbstract:5-Aza-deoxycytidine (5-aza-dC) confers neuroprotection in ischemic mice by inhibiting DNA methylation. Zebularine is another DNA methylation inhibitor, less toxic and more stable in aqueous solutions and, therefore more biologically suitable. We investigated Zebularine’s effects on brain ischemia in a rat middle cerebral artery occlusion (MCAo) model in order to elucidate its therapeutic potential. Male Wistar wild-type (WT) rats were randomly allocated to three treatment groups, vehicle, Zebularine 100 μg, and Zebularine 500 μg. Saline (10 μL) or Zebularine (10 μL) was administered intracerebroventricularly 20 min before 45-min occlusion of the middle cerebral artery. Reperfusion was allowed after 45-min occlusion, and the rats were sacrificed at 24-h reperfusion. The brains were removed, sliced, and stained with 2 % 2,3,5-triphenyltetrazolium chloride (TTC) before measuring infarct size. Zebularine (500 μg) reduced infarct volumes significantly (p
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dna methylation inhibitor Zebularine confersstroke protection in ischemic rats
2015Co-Authors: Hua Dock, Annette Theodorsson, Elvar TheodorssonAbstract:Aza-deoxycytidine (5-aza-dC) confers neuropro- tection in ischemic mice by inhibiting DNA methylation. Zebularine is another DNA methylation inhibitor, less toxic and more stable in aqueous solutions and, therefore more bi- ologically suitable. We investigated Zebularine's effects on brain ischemia in a rat middle cerebral artery occlusion (MCAo) model in order to elucidate its therapeutic potential. Male Wistar wild-type (WT) rats were randomly allocated to three treatment groups, vehicle, Zebularine 100 μg, and Zebularine 500 μg. Saline (10 μL) or Zebularine (10 μL) was administered intracerebroventricularly 20 min before 45-min occlusion of the middle cerebral artery. Reperfusion was allowed after 45-min occlusion, and the rats were sacrificed at 24-h reperfusion. The brains were removed, sliced, and stained with 2 % 2,3,5-triphenyltetrazolium chlo- ride (TTC) before measuring infarct size. Zebularine (500 μg) reduced infarct volumes significantly (p<0.05) by 61 % from 20.7±4.2 % in the vehicle treated to 8.1±1.6 % in the Zebularine treated. Zebularine (100 μg) also reduced infarct volumes dramatically by 55 to 9.4±1.2 %. The mechanisms behind this neuroprotection is not yet known, but the results agree with previous studies and support the notion that Zebularine-induced inhibition of DNA methyltransferase ameliorates ischemic brain injury in rats.
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DNA Methylation Inhibitor Zebularine ConfersStroke Protection in Ischemic Rats
2015Co-Authors: Hua Dock, Annette Theodorsson, Elvar TheodorssonAbstract:Aza-deoxycytidine (5-aza-dC) confers neuropro- tection in ischemic mice by inhibiting DNA methylation. Zebularine is another DNA methylation inhibitor, less toxic and more stable in aqueous solutions and, therefore more bi- ologically suitable. We investigated Zebularine's effects on brain ischemia in a rat middle cerebral artery occlusion (MCAo) model in order to elucidate its therapeutic potential. Male Wistar wild-type (WT) rats were randomly allocated to three treatment groups, vehicle, Zebularine 100 μg, and Zebularine 500 μg. Saline (10 μL) or Zebularine (10 μL) was administered intracerebroventricularly 20 min before 45-min occlusion of the middle cerebral artery. Reperfusion was allowed after 45-min occlusion, and the rats were sacrificed at 24-h reperfusion. The brains were removed, sliced, and stained with 2 % 2,3,5-triphenyltetrazolium chlo- ride (TTC) before measuring infarct size. Zebularine (500 μg) reduced infarct volumes significantly (p
