甲基化抗肿瘤药物治疗中的DNA修复:治疗效果与健康结局

doi:10.3969/j.issn.1004-616x.2016.05.001

摘要/Abstract

摘要： 在甲基化抗肿瘤药物治疗中，DNA修复是决定治疗效果与不良生物效应（如突变、癌变和致畸）的一个关键机制。本文主要对甲基化抗肿瘤药物在DNA修复过程的作用进行综述。尽管这些抗肿瘤药物无选择性地靶向作用于癌细胞和正常细胞的DNA，但因其削弱了癌细胞内某些特异的DNA修复活动，从而更多地杀死癌细胞。单功能的烷化剂显示出甲基化改变特性（丙卡巴肼、达卡巴嗪、链脲佐菌素、替莫唑胺），或者氯乙基化形成单加合物并在下一步反应中引起DNA链内交联（洛莫司汀、尼莫地平、卡莫司汀、福莫司汀）。癌细胞对抗肿瘤药物的一个主要机制是通过自杀酶O⁶-甲基鸟嘌呤-DNA甲基转移酶（MGMT）直接逆转DNA损伤，形成O⁶-甲基鸟嘌呤和O⁶-氯化鸟嘌呤。由于MGMT对恶性肿瘤治疗的结局有显著影响，它被认为是一个耐药的重要标志，特别是在高级恶性胶质瘤。MGMT也被认为是甲基化抗肿瘤药物有效性的预测标志，许多临床试验正在分析MGMT抑制对治疗效果的影响。其他涉及甲基化抗肿瘤药物耐药的DNA修复因素包括错配修复、通过同源重组和DNA双链断裂（DSB）信号启动的相关修复。碱基切除修复和alkB同源蛋白（如ABH2）也可能与烷化类药物耐药有关，该现象在高表达MGMT的细胞株异常明显。对于这些机制的进一步了解，将有助于设计更为有效的治疗方案，同时减少副作用。

关键词: 替莫唑胺, O⁶-甲基鸟嘌呤-DNA甲基转移酶, 烷基转移酶, 启动子甲基化, DNA修复, 肿瘤, 生物标记, 药物抗性, 胶质母细胞瘤

Abstract: For patients who are treated with anti-cancer drugs,DNA repair is often a critical mechanism,which determines therapeutic efficacy and/or adverse biological consequences such as mutagenesis,carcinogenesis and teratogenesis. This brief review is focused onto DNA repair activities on damage resulting from exposure to methylating anticancer drugs. Although these drugs unselectively target DNA from cancer and normal cells,the cancer-specific killing effect is likely due to downregulation of specific DNA repair activities,thereby killing more cancer than normal cells. The monofunctional alkylating agents exhibit methylating properties (procarbazine,dacarbazine,streptozotocine, temozolomide) or they chloroethylate the DNA forming monoadducts and,in a second step,DNA interstrand crosslinks (lomustine,nimustine,carmustine,fotemustine). A major mechanism of defense of cancer cells to these drugs is direct reversal of the critical DNA damage,O⁶-methylguanine as well as O⁶-chloroethylguanine,by the suicide enzyme O⁶-methylguanine-DNA methyltransferase (MGMT),which represents an important drug resistance marker especially in high-grade malignant gliomas. Since MGMT has a significant impact on the outcome of anti-cancer therapy,it is a predictive marker of the effectiveness of methylating anticancer drugs,and clinical trials are underway analyzing the influence of MGMT inhibition on the therapeutic success. Other DNA repair factors involved in methylating drug resistance are mismatch repair,DNA double-strand break (DSB) repair by homologous recombination and DSB signaling. Base excision repair and alkB homologous proteins (such as ABH2) might also contribute to alkylating drug resistance notably in cells expressing a high amount of MGMT. Better understanding of these mechanisms will be helpful in designing more effective therapies that have less adverse outcomes.

Key words: temozolomide, O⁶-methylguanine-DNA methyltransferase, alkyltransferase, promoter methylation, DNA repair, cancer, biomarker, drug resistance, glioblastoma

中图分类号:

R730.5

Bernd Kaina. 甲基化抗肿瘤药物治疗中的DNA修复:治疗效果与健康结局[J]. 癌变·畸变·突变, 2016, 28(5): 333-341,347.

Bernd Kaina. DNA repair in cancer therapy with methylating anticancer drugs: improving therapeutic efficacy and reducing side effects[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(5): 333-341,347.

参考文献

[1] Beranek DT.Distribution of methyl and ethyl adducts following alkylation with monofunctional alkylating agents[J].Mutat Res,1990,231(1):11-30.
[2] Beranek DT,Weis CC,Swenson DH.A comprehensive quantitative analysis of methylated and ethylated DNA using high pressure liquid chromatography[J].Carcinogenesis,1980,1:595-606.
[3] Newlands E,Stevens M,Wedge S,et al.Temozolomide:a review of its discovery,chemical properties,pre-clinical development and clinical trials[J].Cancer Treat Rev,1997,23:35-61.
[4] Pegg AE.Repair of O⁶-alkylguanine by alkyltransferases[J].Mutation Res,2000,462:83-100.
[5] Margison GP,Povey AC,Kaina B,et al.Variability and regulation of O⁶-alkylguanine-DNA alkyltransferase[J].Carcino-genesis,2003,24(4):625-635.
[6] Kaina B,Christmann M,Naumann S,et al.MGMT:key node in the battle against genotoxicity,carcinogenicity and apoptosis induced by alkylating agents[J].DNA Repair (Amst),2007,6(8):1079-1099.
[7] Preuss I,Thust R,Kaina B.Protective effect of 06-methylguanine-DNA methyltransferase (MGMT) on the cytotoxic and recombinogenic activity of different antineoplastic drugs[J].Int J Cancer,1996,65:506-512.
[8] Kaina B,Fritz G,Mitra S,et al.Transfection and expression of human O⁶-methylguanine-DNA methyltransferase (MGMT) cDNA in Chinese hamster cells:the role of MGMT in protection against the genotoxic effects of alkylating agents[J].Carcinogenesis,1991,12:1857-1867.
[9] Kaina B,Margison GP,Christmann M.Targeting O⁶-methylguanine-DNA methyltransferase with specific inhibitors as a strategy in cancer therapy[J].Cell Mol Life Sci,2010,67(21):3663-3681.
[10] Quiros S,Roos WP,Kaina B.Processing of O⁶-methylguanine into DNA double-strand breaks requires two rounds of replication whereas apoptosis is also induced in subsequent cell cycles[J].Cell Cycle,2010,9(1):168-178.
[11] Ochs K,Kaina B.Apoptosis induced by DNA damage O⁶-methylguanine is Bcl-2 and caspase-9/3 regulated and Fas/caspase-8 independent[J].Cancer Res,2000,60(20):5815-5824.
[12] Yoshioka K,Yoshioka Y,Hsieh P.ATR kinase activation mediated by mutsalpha and mutlalpha in response to cytotoxic O⁶-methylguanine adducts[J].Mol Cell,2006,22(4):501-510.
[13] Klapacz J,Meira LB,Luchetti DG,et al.O⁶-methylguanine-induced cell death involves exonuclease 1 as well as DNA mismatch recognition in vivo[J].Proc Natl Acad Sci U S A,2009,106(2):576-581.
[14] Caporali S,Falcinelli S,Starace G,et al.DNA damage induced by temozolomide signals to both ATM and ATR:role of the mismatch repair system[J].Mol Pharmacol,2004,66(3):478-491.
[15] Roos WP,Batista LFZ,Naumann S,et al.Apoptosis in malignant glioma cells triggered by the temozolomide-induced DNA lesion O⁶-methylguanine[J].Oncogene,2007,26:186-197.
[16] Roos WP,Nikolova T,Quiros S,et al.Brca2/Xrcc2 dependent HR,but not NHEJ,is required for protection against O⁶-methylguanine triggered apoptosis,DSBs and chromosomal aberrations by a process leading to SCEs[J].DNA Repair (Amst),2009,8(1):72-86.
[17] Kaina B,Ochs K,Grosch S,et al.BER,MGMT,and MMR in defense against alkylation-induced genotoxicity and apoptosis[J].Prog Nucleic Acid Res Mol Biol,2001,68:41-54.
[18] Eich M,Roos WP,Nikolova T,et al.Contribution of ATM and ATR to the resistance of glioblastoma and malignant melanoma cells to the methylating anticancer drug temozolomide[J].Mol Cancer Ther,2013,12(11):2529-2540.
[19] Stojic L,Mojas N,Cejka P,et al.Mismatch repair-dependent G2 checkpoint induced by low doses of SN1 type methylating agents requires the ATR kinase[J].Genes Dev,2004,18(11):1331-1344.
[20] Kaina B,Ziouta A,Ochs K,et al.Chromosomal instability,reproductive cell death and apoptosis induced by O⁶-methylguanine in Mex-,Mex+and methylation-tolerant mismatch repair compromised cells:facts and models[J].Mutation Res,1997,381:227-241.
[21] Meikrantz W,Bergom MA,Memisoglu A,et al.O⁶-alkyl-guanine DNA lesions trigger apoptosis[J].Carcinogenesis,1998,19:369-372.
[22] Ochs K,Kaina B.Apoptosis induced by DNA damage O⁶-methylguanine is Bcl-2 and caspase-9/-3 regulated and Fas/caspase-8 independent[J].Cancer Res,2000,60:5815-5824.
[23] Tomicic MT,Meise R,Aasland D,et al.Apoptosis induced by temozolomide and nimustine in glioblastoma cells is supported by JNK/c-Jun-mediated induction of the BH3-only protein BIM[J].Oncotarget,2015,6(32):33755-33768.
[24] Christmann M,Verbeek B,Roos WP,et al.O⁶-methylguanine-DNA methyltransferase (MGMT) in normal tissues and tumors:Enzyme activity,promoter methylation and immunohistochemistry[J].Biochim Biophys Acta,2011,1816(2):179-190.
[25] Preuss I,Eberhagen I,Haas S,et al.O⁶-methylguanine-DNA methyltransferase activity in breast and brain tumors[J].Int J Cancer,1995,61(3):321-326.
[26] Christmann M,Nagel G,Horn S,et al.MGMT activity,promoter methylation and immunohistochemistry of pretreatment and recurrent malignant gliomas:a comparative study on astrocytoma and glioblastoma[J].Int J Cancer,2010,127(9):2106-2118.
[27] Dolan ME,Pegg AE,Dumenco LL,et al.Comparison of the inactivation of mammalian and bacterial O⁶-alkylguanine-DNA alkyltransferases by O⁶-benzylguanine and O⁶-methylguanine[J].Carcinogenesis,1991,12(12):2305-2309.
[28] Preuss I,Thust R,Kaina B.Protective effect of O⁶-methylguanine-DNA methyltransferase (MGMT) on the cytotoxic and recombinogenic activity of different antineoplastic drugs[J].Int J Cancer,1996,65(4):506-512.
[29] Pegg AE.Mammalian O⁶-alkylguanine-DNA alkyltransferase:regulation and importance in response to alkylating carcinogenic and therapeutic agents[J].Cancer Res,1990,50(19):6119-6129.
[30] Belanich M,Pastor M,Randall T,et al.Retrospective study of the correlation between the DNA repair protein alkyl-transferase and survival of brain tumor patients treated with carmustine[J].Cancer Res,1996,56(4):783-788.
[31] Jaeckle KA,Eyre HJ,Townsend JJ,et al.Correlation of tumor O⁶-methylguanine-DNA methyltransferase levels with survival of malignant astrocytoma patients treated with bis-chloroethylnitrosourea:a Southwest Oncology Group study[J].J Clin Oncol,1998,16(10):3310-3315.
[32] Mineura K,Izumi I,Watanabe K,et al.Influence of O⁶-methylguanine-DNA methyltransferase activity on chloroethyl-nitrosourea chemotherapy in brain tumors[J].Int J Cancer,1993,55(1):76-81.
[33] Mineura K,Watanabe K,Yanagisawa T,et al.Quantification of O⁶-methylguanine-DNA methyltransferase mRNA in human brain tumors[J].Biochim Biophys Acta,1996,1289(1):105-109.
[34] Fabi A,Metro G,Russillo M,et al.Treatment of recurrent malignant gliomas with fotemustine monotherapy:impact of dose and correlation with MGMT promoter methylation[J].BMC Cancer,2009,9:101.
[35] Hegi ME,Diserens AC,Gorlia T,et al.MGMT gene silencing and benefit from temozolomide in glioblastoma[J].N Engl J Med,2005,352(10):997-1003.
[36] Hegi ME,Diserens AC,Godard S,et al.Clinical trial substantiates the predictive value of O⁶-methylguanine-DNA methyltransferase promoter methylation in glioblastoma patients treated with temozolomide[J].Clin Cancer Res,2004,10(6):1871-1874.
[37] Friedman HS,Kerby T,Calvert H.Temozolomide and treatment of malignant glioma[J].Clin Cancer Res,2000,6(7):2585-2597.
[38] Friedman HS,Kokkinakis DM,Pluda J,et al.Phase I trial of O⁶-benzylguanine for patients undergoing surgery for malignant glioma[J].J Clin Oncol,1998,16(11):3570-3575.
[39] Stupp R,Hegi ME,Mason WP,et al.Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase Ⅲ study:5-year analysis of the EORTC-NCIC trial[J].Lancet Oncol,2009,10(5):459-466.
[40] Wiewrodt D,Nagel G,Dreimuller N,et al.MGMT in primary and recurrent human glioblastomas after radiation and chemotherapy and comparison with p53 status and clinical outcome[J].Int J Cancer,2008,122(6):1391-1399.
[41] Christmann M,Verbeek B,Roos WP,et al.O⁶-methylguanine-DNA methyltransferase (MGMT) in normal tissues and tumors:Enzyme activity,promoter methylation and immunohistochemistry[J].Biochim Biophys Acta,2011,1816(2):179-190.
[42] Krokan H,Haugen A,Myrnes B,et al.Repair of premutagenic DNA lesions in human fetal tissues:evidence for low levels of O⁶-methylguanine-DNA methyltransferase and uracil-DNA glycosylase activity in some tissues[J].Carcinogenesis,1983,4:1559-1564.
[43] Esteller M,Garcia-Foncillas J,Andion E,et al.Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents[J].N Engl J Med,2000,343(19):1350-1354.
[44] Everhard S,Kaloshi G,Criniere E,et al.MGMT methylation:a marker of response to temozolomide in low-grade gliomas[J].Ann Neurol,2006,60(6):740-743.
[45] Mollemann M,Wolter M,Felsberg J,et al.Frequent promoter hypermethylation and low expression of the MGMT gene in oligodendroglial tumors[J].Int J Cancer,2005,113(3):379-385.
[46] Switzeny OJ,Christmann M,Renovanz M,et al.MGMT promoter methylation determined by HRM in comparison to MSP and pyrosequencing for predicting high-grade glioma response[J].Clin Epigenetics,2016,8:49.
[47] Esteller M,Hamilton SR,Burger PC,et al.Inactivation of the DNA repair gene O⁶-methylguanine-DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia[J].Cancer Res,1999,59(4):793-797.
[48] Shibata T,Glynn N,McMurry TB,et al.Novel synthesis of O⁶-alkylguanine containing oligodeoxyribonucleotides as substrates for the human DNA repair protein,O⁶-methylguanine DNA methyltransferase (MGMT)[J].Nucleic Acids Res,2006,34(6):1884-1891.
[49] Felker GM,Friedman HS,Dolan ME,et al.Treatment of subcutaneous and intracranial brain tumor xenografts with O⁶-benzylguanine and 1,3-bis (2-chloroethyl)-1-nitrosourea[J].Cancer Chemother Pharmacol,1993,32(6):471-476.
[50] Friedman HS,Dolan ME,Moschel RC,et al.Enhancement of nitrosourea activity in medulloblastoma and glioblastoma multiforme[J].J Natl Cancer Inst,1992,84(24):1926-1931.
[51] Rhines LD,Sampath P,Dolan ME,et al.O⁶-benzylguanine potentiates the antitumor effect of locally delivered carmustine against an intracranial rat glioma[J].Cancer Res,2000,60(22):6307-6310.
[52] Marathi UK,Dolan ME,Erickson LC.Anti-neoplastic activity of sequenced administration of O⁶-benzylguanine,streptozotocin,and 1,3-bis (2-chloroethyl)-1-nitrosourea in vitro and in vivo[J].Biochem Pharmacol,1994,48(11):2127-2134.
[53] Wan Y,Wu D,Gao H,et al.Potentiation of BCNU anticancer activity by O⁶-benzylguanine:a study in vitro and in vivo[J].J Environ Pathol Toxicol Oncol,2000,19(1/2):69-75.
[54] Kokkinakis DM,Ahmed MM,Chendil D,et al.Sensitization of pancreatic tumor xenografts to carmustine and temozolomide by inactivation of their O⁶-methylguanine-DNA methyltransferase with O⁶-benzylguanine or O⁶-benzyl-2'-deoxyguanosine[J].Clin Cancer Res,2003,9(10 Sup 1):3801-3807.
[55] Friedman HS,Keir S,Pegg AE,et al.O⁶-benzylguanine-mediated enhancement of chemotherapy[J].Mol Cancer Ther,2002,1(11):943-948.
[56] Wagner LM,McLendon RE,Yoon KJ,et al.Targeting methylguanine-DNA methyltransferase in the treatment of neuroblastoma[J].Clin Cancer Res,2007,13(18 Sup 1):5418-5425.
[57] Middleton MR,Kelly J,Thatcher N,et al.O⁶-(4-bromothenyl) guanine improves the therapeutic index of temozolomide against A375M melanoma xenografts[J].Int J Cancer,2000,85(2):248-252.
[58] Middleton MR,Kelly J,Goodger S,et al.Four-hourly scheduling of temozolomide improves tumour growth delay but not therapeutic index in A375M melanoma xenografts[J].Cancer Chemother Pharmacol,2000,45(1):15-20.
[59] Middleton MR,Thatcher N,McMurry TB,et al.Effect of O⁶-(4-bromothenyl) guanine on different temozolomide schedules in a human melanoma xenograft model[J].Int J Cancer,2002,100(5):615-617.
[60] Clemons M,Kelly J,Watson AJ,et al.O⁶-(4-bromothenyl) guanine reverses temozolomide resistance in human breast tumour MCF-7 cells and xenografts[J].Br J Cancer,2005,93(10):1152-1156.
[61] Quinn JA,Pluda J,Dolan ME,et al.Phase Ⅱ trial of carmustine plus O⁶-benzylguanine for patients with nitrosourea-resistant recurrent or progressive malignant glioma[J].J Clin Oncol,2002,20(9):2277-2283.
[62] Koch D,Hundsberger T,Boor S,et al.Local intracerebral administration of O⁶-benzylguanine combined with systemic chemotherapy with temozolomide of a patient suffering from a recurrent glioblastoma[J].J Neurooncol,2007,82:85-89.
[63] Javanmard S,Loktionova NA,Fang Q,et al.Inactivation of O⁶-alkylguanine-DNA alkyltransferase by folate esters of O⁶-benzyl-2'-deoxyguanosies and of O⁶-(4-(hydroxymethyl) benzyl) guanine[J].J Med Chem,2007,50(21):5193-5201.
[64] Yamamoto T,Seino Y,Fukumoto H,et al.Over-expression of facilitative glucose transporter genes in human cancer[J].Biochem Biophys Res Commun,1990,170(1):223-230.
[65] Reinhard J,Eichhorn U,Wiessler M,et al.Inactivation of O⁶-methylguanine-DNA methyltransferase by glucose-conjugated inhibitors[J].Int J Cancer,2001,93(3):373-379.
[66] Reinhard J,Hull WE,von der Lieth CW,et al.Monosaccharide-linked inhibitors of O⁶-methylguanine-DNA methyltransferase (MGMT):synthesis,molecular modeling,and structure-activity relationships[J].J Med Chem,2001,44(24):4050-4061.
[67] Kaina B,Muhlhausen U,Piee-Staffa A,et al.Inhibition of O⁶-methylguanine-DNA methyltransferase by glucose-conjugated inhibitors:comparison with nonconjugated inhibitors and effect on fotemustine and temozolomide-induced cell death[J].J Pharmacol Exp Ther,2004,311(2):585-593.
[68] Tomaszowski KH,Schirrmacher R,Kaina B.Multidrug efflux pumps attenuate the effect of mgmt inhibitors[J].Mol Pharm,2015,12(11):3924-3934.
[69] Roth RB,Samson LD.Gene transfer to suppress bone marrow alkylation sensitivity[J].Mut Res,2000,462(2/3):107-120.
[70] Adair JE,Beard BC,Trobridge GD,et al.Extended survival of glioblastoma patients after chemoprotective HSC gene therapy[J].Sci Transl Med,2012,4(133):133-157.