DNMT3a plays a role in switches between doxorubicin-induced senescence and apoptosis of colorectal cancer cells
Yu Zhang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorYanyan Gao
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorGuoping Zhang
Biology Section of Guangdong Medical College, Dongguan, Guangdong, China
Search for more papers by this authorShuyan Huang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorZhixiong Dong
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorChenfei Kong
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorDongmei Su
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorJuan Du
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorShan Zhu
Biology Section of Guangdong Medical College, Dongguan, Guangdong, China
Search for more papers by this authorQian Liang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorJianchao Zhang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorCorresponding Author
Jun Lu
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Tel.: +86-431-85098729
Jun Lu, The Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, Changchun 130024, China
Baiqu Huang, The Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
Search for more papers by this authorCorresponding Author
Baiqu Huang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Tel.: +86-431-85099798, Fax: +86-431-85099768
Jun Lu, The Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, Changchun 130024, China
Baiqu Huang, The Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
Search for more papers by this authorYu Zhang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorYanyan Gao
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorGuoping Zhang
Biology Section of Guangdong Medical College, Dongguan, Guangdong, China
Search for more papers by this authorShuyan Huang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorZhixiong Dong
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorChenfei Kong
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorDongmei Su
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorJuan Du
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorShan Zhu
Biology Section of Guangdong Medical College, Dongguan, Guangdong, China
Search for more papers by this authorQian Liang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorJianchao Zhang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Search for more papers by this authorCorresponding Author
Jun Lu
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Tel.: +86-431-85098729
Jun Lu, The Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, Changchun 130024, China
Baiqu Huang, The Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
Search for more papers by this authorCorresponding Author
Baiqu Huang
The Institute of Genetics and Cytology, The Key Laboratory of Molecular Epigenetics of Ministry of Education (MOE), Northeast Normal University, Changchun, China
Tel.: +86-431-85099798, Fax: +86-431-85099768
Jun Lu, The Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, Changchun 130024, China
Baiqu Huang, The Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China
Search for more papers by this authorAbstract
The DNA-damaging drug doxorubicin (Dox) induces cell senescence at concentrations significantly lower than those required for induction of apoptosis. At low Dox concentrations, tumor suppressor p53 is activated, which enhances the expression of p21Waf1/Cip1 (p21). At high concentrations, Dox activates p53 leading to apoptosis without enhancing p21 expression. The underlying mechanisms and factors that govern the differential effects of Dox in inducing senescence and apoptosis are unclear. Here, we report that the DNA methyltransferase (DNMT) DNMT3a was upregulated by Dox especially at concentrations that induced apoptosis in HCT116 colorectal cancer cells, and this process was regulated by p53. Meanwhile, p21 expression was significantly upregulated at senescence-inducing concentrations and kept low on treatment with apoptosis-inducing concentrations of Dox. The differential expression of DNMT3a and p21 in response to Dox suggests that DNMT3a may be a key factor in switches between Dox-induced senescence and apoptosis. Moreover, when DNMT3a was silenced, treatment of HCT116 cells with apoptosis-inducing concentration of Dox increased the percentage of cells undergoing senescence, accompanied by upregulation of p21. Contrarily, senescence-inducing concentration of Dox promoted apoptosis rate, and p21 expression was repressed. Surprisingly, no changes in DNA methylation status at p21 promoter were detected at either ranges of Dox, although DNMT3a and HDAC1 were recruited to p21 promoter at apoptosis-inducing Dox concentration, where they were present in the same complex. Overall, these data demonstrate that DNMT3a impacts the expression of p21 and plays a role in determining the Dox-induced senescence and apoptosis in HCT116 cells.
References
- 1 Gewirtz DA. A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin. Biochem Pharmacol 1999; 57: 727–41.
- 2 Rebbaa A, Zheng X, Chou PM, Mirkin BL. Caspase inhibition switches doxorubicin-induced apoptosis to senescence. Oncogene 2003; 22: 2805–11.
- 3 El-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE, Wang Y, et al. WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 1994; 54: 1169–74.
- 4 Chang BD, Swift ME, Shen M, Fang J, Broude EV, Roninson IB. Molecular determinants of terminal growth arrest induced in tumor cells by a chemotherapeutic agent. Proc Natl Acad Sci USA 2002; 99: 389–94.
- 5 Schuler M, Green DR. Transcription, apoptosis and p53: catch-22. Trends Genet 2005; 21: 182–7.
- 6 Roninson IB. Oncogenic functions of tumour suppressor p21(Waf1/Cip1/Sdi1): association with cell senescence and tumour-promoting activities of stromal fibroblasts. Cancer Lett 2002; 179: 1–14.
- 7 Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach D. p21 is a universal inhibitor of cyclin kinases. Nature 1993; 366: 701–4.
- 8 Deng C, Zhang P, Harper JW, Elledge SJ, Leder P. Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 1995; 82: 675–84.
- 9 Mahyar-Roemer M, Roemer K. p21 Waf1/Cip1 can protect human colon carcinoma cells against p53-dependent and p53-independent apoptosis induced by natural chemopreventive and therapeutic agents. Oncogene 2001; 20: 3387–98.
- 10 Bunz F, Hwang PM, Torrance C, Waldman T, Zhang Y, Dillehay L, Williams J, Lengauer C, Kinzler KW, Vogelstein B. Disruption of p53 in human cancer cells alters the responses to therapeutic agents. J Clin Invest 1999; 104: 263–9.
- 11 Martinez LA, Yang J, Vazquez ES, Rodriguez-Vargas Mdel C, Olive M, Hsieh JT, Logothetis CJ, Navone NM. p21 modulates threshold of apoptosis induced by DNA-damage and growth factor withdrawal in prostate cancer cells. Carcinogenesis 2002; 23: 1289–96.
- 12 Wu S, Cetinkaya C, Munoz-Alonso MJ, von der Lehr N, Bahram F, Beuger V, Eilers M, Leon J, Larsson LG. Myc represses differentiation-induced p21CIP1 expression via Miz-1-dependent interaction with the p21 core promoter. Oncogene 2003; 22: 351–60.
- 13 Jin YH, Yoo KJ, Lee YH, Lee SK. Caspase 3-mediated cleavage of p21WAF1/CIP1 associated with the cyclin A-cyclin-dependent kinase 2 complex is a prerequisite for apoptosis in SK-HEP-1 cells. J Biol Chem 2000; 275: 30256–63.
- 14 Rebbaa A, Zheng X, Chu F, Mirkin BL. The role of histone acetylation versus DNA damage in drug-induced senescence and apoptosis. Cell Death Differ 2006; 13: 1960–7.
- 15 Bestor TH. The DNA methyltransferases of mammals. Hum Mol Genet 2000; 9: 2395–402.
- 16 Okano M, Li E. Genetic analyses of DNA methyltransferase genes in mouse model system. J Nutr 2002; 132: 2462S–5S.
- 17 Okano M, Bell DW, Haber DA, Li E. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 1999; 99: 247–57.
- 18 Geiman TM, Sankpal UT, Robertson AK, Zhao Y, Zhao Y, Robertson KD. DNMT3B interacts with hSNF2H chromatin remodeling enzyme. HDACs 1 and 2, and components of the histone methylation system. Biochem Biophys Res Commun 2004; 318: 544–55.
- 19 Fuks F, Burgers WA, Godin N, Kasai M, Kouzarides T. Dnmt3a binds deacetylases and is recruited by a sequence-specific repressor to silence transcription. Embo J 2001; 20: 2536–44.
- 20 Robertson KD, Uzvolgyi E, Liang G, Talmadge C, Sumegi J, Gonzales FA, Jones PA. The human DNA methyltransferases (DNMTs) 1, 3a and 3b: coordinate mRNA expression in normal tissues and overexpression in tumors. Nucleic Acids Res 1999; 27: 2291–8.
- 21 Schneider-Stock R, Ocker M. Epigenetic therapy in cancer: molecular background and clinical development of histone deacetylase and DNA methyltransferase inhibitors. IDrugs 2007; 10: 557–61.
- 22 Yokochi T, Robertson KD. Doxorubicin inhibits DNMT1, resulting in conditional apoptosis. Mol Pharmacol 2004; 66: 1415–20.
- 23 Su D, Zhu S, Han X, Feng Y, Huang H, Ren G, Pan L, Zhang Y, Lu J, Huang B. BMP4-Smad signaling pathway mediates adriamycin-induced premature senescence in lung cancer cells. J Biol Chem 2009; 284: 12153–64.
- 24 Jiemjit A, Fandy TE, Carraway H, Bailey KA, Baylin S, Herman JG, Gore SD. p21(WAF1/CIP1) induction by 5-azacytosine nucleosides requires DNA damage. Oncogene 2008; 27: 3615–23.
- 25 Lu R, Wang X, Chen ZF, Sun DF, Tian XQ, Fang JY. Inhibition of the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway decreases DNA methylation in colon cancer cells. J Biol Chem 2007; 282: 12249–59.
- 26 Zhang Y, Huang S, Dong W, Li L, Feng Y, Pan L, Han Z, Wang X, Ren G, Su D, Huang B, Lu J. SOX7, down-regulated in colorectal cancer, induces apoptosis and inhibits proliferation of colorectal cancer cells. Cancer Lett 2009; 277: 29–37.
- 27 Kim TK, Lee JS, Jung JE, Oh SY, Kwak S, Jin X, Lee SY, Lee JB, Chung YG, Choi YK, You S, Kim H. Interferon regulatory factor 3 activates p53-dependent cell growth inhibition. Cancer Lett 2006; 242: 215–21.
- 28 Wang X, Pan L, Feng Y, Wang Y, Han Q, Han L, Han S, Guo J, Huang B, Lu J. P300 plays a role in p16(INK4a) expression and cell cycle arrest. Oncogene 2008; 27: 1894–904.
- 29 Chang BD, Broude EV, Dokmanovic M, Zhu H, Ruth A, Xuan Y, Kandel ES, Lausch E, Christov K, Roninson IB. A senescence-like phenotype distinguishes tumor cells that undergo terminal proliferation arrest after exposure to anticancer agents. Cancer Res 1999; 59: 3761–7.
- 30 Wang YA, Kamarova Y, Shen KC, Jiang Z, Hahn MJ, Wang Y, Brooks SC. DNA methyltransferase-3a interacts with p53 and represses p53-mediated gene expression. Cancer Biol Ther 2005; 4: 1138–43.
- 31 Wells SI, Francis DA, Karpova AY, Dowhanick JJ, Benson JD, Howley PM. Papillomavirus E2 induces senescence in HPV-positive cells via pRB- and p21(CIP)-dependent pathways. Embo J 2000; 19: 5762–71.
- 32 Vousden KH, Lu X. Live or let die: the cell's response to p53. Nat Rev Cancer 2002; 2: 594–604.
- 33 Roninson IB. Tumor cell senescence in cancer treatment. Cancer Res 2003; 63: 2705–15.
- 34 Li CH, Tzeng SL, Cheng YW, Kang JJ. Chloramphenicol-induced mitochondrial stress increases p21 expression and prevents cell apoptosis through a p21-dependent pathway. J Biol Chem 2005; 280: 26193–9.
- 35 Jair KW, Bachman KE, Suzuki H, Ting AH, Rhee I, Yen RW, Baylin SB, Schuebel KE. De novo CpG island methylation in human cancer cells. Cancer Res 2006; 66: 682–92.
- 36 Deplus R, Brenner C, Burgers WA, Putmans P, Kouzarides T, de Launoit Y, Fuks F. Dnmt3L is a transcriptional repressor that recruits histone deacetylase. Nucleic Acids Res 2002; 30: 3831–8.
- 37 Di Croce L, Raker VA, Corsaro M, Fazi F, Fanelli M, Faretta M, Fuks F, Lo Coco F, Kouzarides T, Nervi C, Minucci S, Pelicci PG. Methyltransferase recruitment and DNA hypermethylation of target promoters by an oncogenic transcription factor. Science 2002; 295: 1079–82.
- 38 Brenner C, Deplus R, Didelot C, Loriot A, Vire E, de Smet C, Gutierrez A, Danovi D, Bernard D, Boon T, Pelicci PG, Amati B, et al. Myc represses transcription through recruitment of DNA methyltransferase corepressor. Embo J 2005; 24: 336–46.
Citing Literature
