The role of XPC protein deficiency in tobacco smoke-induced DNA hypermethylation of tumor suppressor genes


DNA hypermethylation of tumor suppressor genes has been frequently observed in cancer patients, and therefore, may provide a valuable biomarker for cancer prevention and treatment. DNA hypermethylation may also provide an important mechanism in cancer progression. Lung cancer is strongly associated with exposure to environmental carcinogens, especially tobacco smoke. DNA damage generated by tobacco smoke is believed to play an important role in lung cancer development. XPC is a DNA damage recognition protein required for DNA repair and other DNA damage responses and attenuated XPC protein levels have been detected in many lung cancer patients. We studied the role of XPC protein deficiency in tobacco smoke-caused DNA hypermethylation of important tumor suppressor genes. Using both normal human fibroblasts (NF) and XPC-deficient hu man fibroblasts (XPC), our DNA methylation studies demonstrated that the XPC deficiency caused elevated levels of DNA hypermethylation in both Brca1 and Mlh1 tumor suppressor genes following exposure to tobacco smoke condensate (TSC). The results of our ChIP studies revealed that the XPC deficiency led to an increased binding of DNA methyltransferase 3A (DNMT3A) at the promoter region CpG island-containing sequences of these genes under the TSC treatment; however, this increase was partially diminished with prior treatment with caffeine. The results of our immuno-precipitation (IP) studies demonstrated a protein-protein interaction of the ATR with DNMT3A. Our western blots revealed that the

XPC deficiency caused an increase in TSC-induced ATR phosphorylation at serine 428, an indicator of ATR activation. All these results suggest that XPC deficiency causes an accelerated DNA hypermethylation in important tumor suppressor genes under tobacco smoke exposure and activation of the ATR signaling pathway is involved in this DNA hypermethylation process.

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Wang, G. , Wang, L. , Bhoopalan, V. , Xi, Y. , Bhalla, D. , Wang, D. and Xu, X. (2013) The role of XPC protein deficiency in tobacco smoke-induced DNA hypermethylation of tumor suppressor genes. Open Journal of Genetics, 3, 285-293. doi: 10.4236/ojgen.2013.34032.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] [1] Kerr, K.M., Galler, J.S., Hagen, J.A., Laird, P.W. and Laird-Offringa, I.A. (2007) The role of DNA methylation in the development and progression of lung adenocarcinoma. Disease Markers, 23, 5-30.
[2] Anglim, P.P., Alonzo, T.A. and Laird-Offringa, I.A. (2008) DNA methylation-based biomarkers for early detection of non-small cell lung cancer: An update. Molecular Cancer, 7, 81.
[3] Risch, A. and Plass, C. (2008) Lung cancer epigenetics and genetics. International Journal of Cancer, 123, 1-7.
[4] Pfeifer, G.P. and Rauch, T.A. (2009) DNA methylation patterns in lung carcinomas. Seminars in Cancer Biology, 19, 181-187.
[5] Van Den Broeck, A., Ozenne, P., Eymin, B. and Gazzeri, S. (2010) Lung cancer: A modified epigenome. Cell Adhesion & Migration, 4, 107-113.
[6] Holland-Frei (2006) Cancer medicine 7. American Association for Cancer Research.
[7] Akhavan-Niaki, H. and Samadani, A.A. (2013) DNA methylation and cancer development: Molecular mechanism. Cell Biochemistry and Biophysics.
[8] Das, P.M. and Singal, R. (2004) DNA methylation and cancer. Journal of Clinical Oncology, 22, 4632-4642.
[9] Sancar, A., Lindsey-Boltz, L.A., Kang, T.H., Reardon, J.T., Lee, J.H. and Ozturk, N. (2010) Circadian clock control of the cellular response to DNA damage. FEBS Letters, 584.2618-2625.
[10] Friedberg, E.C., Walker, G.C., Siede, W., Wood, R.D., Schultz, R.A. and Ellenberger, T. (2006) DNA repair and mutagenesis. ASM Press, Washington DC.
[11] Hanawalt, P. (2002) Subpathways of nucleotide excision repair and their regulation. Oncogene, 21, 8949-8956.
[12] Sarker, A.H., Tsutakawa, S.E., Kostek, S., Ng, C., Shin, D.S., Peris, M., Campeau, E., Tainer, J.A., Nogales, E., and Cooper, P.K. (2005) Recognition of RNA polymerase II and transcription bubbles by XPG, CSB, and TFIIH: Insights for transcription-coupled repair and Cockayne Syndrome. Molecular Cell, 20, 187-198.
[13] Lainé, J.P. and Egly, J.M. (2006) Initiation of DNA repair mediated by a stalled RNA polymerase IIO. The EMBO Iournal, 25, 387-397.
[14] Wood, R.D. (1999) DNA damage recognition during nucleotide excision repair in mammalian cells. Biochimie, 81, 39-44.
[15] Sugasawa, K., Okamoto, T., Shimizu, Y., Masutani, C., Iwai, S. and Hanaoka, F. (2001) A multistep damage recognition mechanism for global genomic nucleotide excision repair. Genes & Development, 15, 507-521.
[16] Wang, G., Chuang, L., Zhang, X., Colton, S., Dombkowski, A., Reiners, J., Diakiw, A. and Xu, X.S. (2004) The initiative role of XPC protein in cisplatin DNA damaging treatment-mediated cell cycle regulation. Nucleic Acids Research, 32, 2231-2240.
[17] Colton, S.L., Xu, X., Wang, Y.A. and Wang, G. (2006) The involvement of ataxia-telangiectasia mutated protein activation in nucleotide excision repair-facilitated cell survival with cisplatin treatment. The Journal of Biological Chemistry, 281, 27117-27125.
[18] Lomonaco, S.L., Xu, X. and Wang, G. (2009) The role of Bcl-x(L) protein in nucleotide excision repair-facilitated cell protection against cisplatin-induced apoptosis. DNA and Cell Biology, 28, 285-294.
[19] Hollander, M.C., Philburn, R.T., Patterson, A.D., VelascoMiguel, S., Friedberg, E.C., Linnoila, R.I. and Fornace Jr., A.J. (2005) Deletion of XPC leads to lung tumors in mice and is associated with early events in human lung carcinogenesis. Proceedings of the National Academy of Sciences of the United States of America, 102, 13200-13205.
[20] Chen, Z., Yang, J., Wang, G., Song, B., Li, J. and Xu, Z. (2007) Attenuated expression of xeroderma pigmentosum group C is associated with critical events in human bladder cancer carcinogenesis and progression. Cancer Research, 67, 4578-4585.
[21] Wu, Y.H., Tsai Chang, J.H., Cheng, Y.W., Wu, T.C., Chen, C.Y. and Lee, H. (2007) Xeroderma pigmentosum group C gene expression is predominantly regulated by promoter hypermethylation and contributes to p53 mutation in lung cancers. Oncogene, 26, 4761-4773.
[22] Cheo, D.L., Burns, D.K., Meira, L.B., Houle, J.F. and Friedberg, E.C. (1999) Mutational inactivation of the xeroderma pigmentosum group C gene confers predisposition to 2-acetylaminofluorene-induced liver and lung cancer and to spontaneous testicular cancer in Trp53-/-mice. Cancer Research, 59, 771-775.
[23] Friedberg, E.C., Bond, J.P., Burns, D.K., Cheo, D.L., Greenblatt, M.S., Meira, L.B., Nahari, D. and Reis, A.M. (2000) Defective nucleotide excision repair in Xpc mutant mice and its association with cancer predisposition. Mutation Research/DNA Repair, 459, 99-108.
[24] Wang, Y.C., Lu, Y.P., Tseng, R.C., Lin, R.K., Chang, J.W., Chen, J.T., Shih, C.M. and Chen, C.Y. (2003) Inactivation of hMLH1 and hMSH2 by promoter methylation in primary non-small cell lung tumors and matched sputum samples. The Journal of Clinical Investigation, 111, 887-895.
[25] Okuda, T., Kawakami, K., Ishiguro, K., Oda, M., Omura, K. and Watanabe, G. (2005) The profile of hMLH1 methylation and microsatellite instability in colorectal and non-small cell lung cancer. International Journal of Molecular Medicine, 15, 85-90.
[26] Lee, M.N., Tsen, R.C., Hsu, H.S., Chen, J.Y., Tzao, C., Ho, W.L. and Wang, Y.C. (2007) Epigenetic inactivation of the chromosomal stability control genes BRCA1, BRCA2, and XRCC5 in non-small cell lung cancer. Clinical Cancer Research, 13, 832-838.
[27] Seng, T.J., Currey, N., Cooper, W.A., Lee, C.S., Chan, C., Horvath, L., Sutherland, R.L., Kennedy, C., McCaughan, B. and Kohonen-Corish, M.R. (2008) DLEC1 and MLH1 promoter methylation are associated with poor prognosis in non-small cell lung carcinoma. British Journal of Cancer, 99, 375-382.
[28] Geng, X., Wang, F., Zhang, L. and Zhang, W.M. (2009) Loss of heterozygosity combined with promoter hypermethylation, the main mechanism of human MutL Homolog (hMLH1) gene inactivation in non-small cell lung cancer in a Chinese population. Tumori, 95, 488-494.
[29] Okano, M., Bell, D.W., Haber, D.A. and Li, E. (1999) DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell, 99, 247-257.
[30] Chen, T., Ueda, Y., Dodge, J.E., Wang, Z. and Li, E. Establishment and maintenance of genomic methylation patterns in mouse embryonic stem cells by Dnmt3a and Dnmt3b. Molecular and Cellular Biology, 2003, 23, 5594-5605.
[31] Watanabe, D., Suetake, I., Tada, T. and Tajima, S. (2002) Stage- and cell-specific expression of Dnmt3a and Dnmt3b during embryogenesis, Mechanisms of Development, 118, 187-190.
[32] Tibbetts, R.S., Brumbaugh, K.M., Williams, J.M., Sarkaria, J.N., Cliby, W.A., Shieh, S.Y., Taya, Y., Prives, C. and Abraham, R.T. (1999) A role for ATR in the DNA damage-induced phosphorylation of p53. Genes & Development, 13, 152-157.
[33] Heffernan, T.P., Simpson, D.A., Frank, A.R., Heinloth, A.N., Paules, R.S., Cordeiro-Stone, M. and Kaufmann, W.K. (2002) An ATR- and Chk1-dependent S checkpoint inhibits replicon initiation following UVC-induced DNA damage. Molecular and Cellular Biology, 22, 8552-8561.
[34] Helt, C.E., Cliby, W.A., Keng, P.C., Bambara, R.A. and O’Reilly, M.A. (2005) Ataxia telangiectasia mutated (ATM) and ATM and Rad3-related protein exhibit selective target specificities in response to different forms of DNA damage. The Journal of Biological Chemistry, 280, 1186-1192.
[35] Cimprich, K.A. and Cortez, D. (2008) ATR: An essential regulator of genome integrity. Nature Reviews Molecular Cell Biology, 9, 616-627.
[36] Despras, E., Daboussi, F., Hyrien, O., Marheineke, K. and Kannouche, P.L. (2010) ATR/Chk1 pathway is essential for resumption of DNA synthesis and cell survival in UV-irradiated XP variant cells. Human Molecular Genetics, 19, 1690-1701.
[37] Stiff, T., Walker, S.A., Cerosaletti, K., Goodarzi, A.A., Petermann, E., Concannon, P., O'Driscoll, M., Jeggo, P.A. (2006) ATR-dependent phosphorylation and activation of ATM in response to UV treatment or replication fork stalling. The EMBO Journal, 25, 5775-5782.
[38] Ward, I.M., Minn, K. and Chen, J. (2004) UV-induced ataxia-telangiectasia-mutated and Rad3-related (ATR) activation requires replication stress. The Journal of Biological Chemistry, 279, 9677-9680.
[39] Hervouet, E., Vallette, F.M. and Cartron, P.F. (2009) Dnmt3/transcription factor interactions as crucial players in targted DNA methylation. Epigenetics, 4, 487-499.
[40] Le, May, N., Mota-Fernandes, D., Velez-Cruz, R., Iltis, I., Biard, D. and Egly, J.M. (2010) NER factors are recruited to active promoters and facilitate chromatin modification for transcription in the absence of exogenous genotoxic attack. Molecular and Cellular Biology, 38, 54-66.

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