药物相关性分子靶标检测在儿童恶性实体肿瘤个体化治疗中的初步研究

摘要

目的 初步探讨药物相关性分子靶标检测在儿童恶性实体肿瘤个体化治疗中的临床意义.方法 选择2014年2月至2017年3月期间于重庆医科大学附属儿童医院诊治的150例恶性实体肿瘤患儿,取其手术肿瘤组织标本150份,通过免疫组织化学(IHC)或聚合酶链式反应(PCR)测序的方法检测肿瘤药物相关性分子靶标的表达或突变,并根据靶标对应的药物分别比较间叶来源和非间叶来源的肿瘤,霍奇金淋巴瘤(Hodgkin lymphoma,HL)和非霍奇金淋巴瘤(non-Hodgkin lymphoma,NHL),神经母细胞瘤(neuroblastoma,NB)和肝母细胞瘤(hepatoblastoma,HB)对常见化疗药物的敏感性及毒副作用有无差异.结果 ①共检测肿瘤标本化疗药物相关性分子靶标15个,包括:人切除修复交叉互补基因1 (ERCC1),DNA拓扑异构酶I(TOPO I),DNA拓扑异构酶ⅡA(TOPOⅡA),DNA修复蛋白O6-甲基转移酶(MGMT),微管蛋白β3(tubulinβ3),胸苷酸合成酶(TS),CYP2C19*2,二氢叶酸还原酶[DHFR(C829T)],尿苷二磷酸-葡萄糖醛酸基转移酶(lUGT1 A1* 28),CYP2B6*6,亚甲基四氢叶酸还原酶[MTHFR(C677T)],巯嘌呤甲基转移酶[TPMT(A719G)],TPMT(G238C),二氢嘧啶脱氢酶[DPYD* 2A(14+ 1G＞A)]和DPYD* 9(T85C)基因的表达或多态性.②根据靶标对应的相关药物进行分析,得出常用不同化疗药物在儿童恶性实体肿瘤中的总体敏感率及毒副反应情况:甲氨蝶呤敏感占100.0％,替莫唑胺/卡莫司汀/司莫司汀敏感占66.7％,蒽环类/依托泊苷、氟尿嘧啶敏感占52.2％,铂类敏感占52.1％,长春碱类敏感占51.6％,伊立替康/拓扑替康不敏感占81.5％,环磷酰胺不敏感占53.7％;甲氨蝶呤毒副反应弱占90.0％,巯嘌呤毒副反应弱占87.5％,环磷酰胺毒副反应弱占73.8％,氟尿嘧啶毒副反应弱占64.3％,伊立替康/依托泊苷毒副反应弱占55.6％.③不同来源、不同病理类型的儿童恶性实体肿瘤对同一化疗药物敏感性的差异比较:间叶源性肿瘤较非间叶源性肿瘤对伊立替康/拓扑替康敏感性较高(P＜0.001),NHL较HL对蒽环类/依托泊苷敏感性较高(P ＜0.001),HB较NB对铂类(P=0.011)和长春碱类敏感性较高(P=0.018),差异均有统计学意义.结论 本研究系统性检测了儿童常见恶性实体肿瘤药物相关性分子靶标的表达或多态性,根据检测结果预测药物敏感性及毒副作用,为儿童恶性实体肿瘤化疗方案的制定提供了循证学依据及进一步为个体化用药提供了指导.%Objective To preliminarily explore the clinical significance of drug-related genomic markers for individualized therapy in children with common malignant solid tumors.Methods Surgical specimens in 150 children with malignant solid tumor were collected between February 2014 and March 2017.The drug-related genomic markers were examined by immunohistochemical staining and polymerase chain reaction sequencing.According to identification of genomic markers,we compared the differences of sensitivity and toxicity to common chemotherapeutic agents between mesenchymal and non-mesenchymal tumors,Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL),neuroblastoma (NB) and hepatoblastoma (HB) respectively.Results ①Fifteen genomic markers of anti-neoplastic agents correlations were examined,including gene expression or mutation of excision repair cross-complementing 1 (ERCC1),topoisomerase I (TOPOI),topoisomerase Ⅱ A (TOPO Ⅱ A),O6-methylguanine-DNA methyhransferase (MGMT),tubulin β33,thymidylate synthase (TS),CYP2C19 * 2,dihydrofolate reductase (DHFR C829T),UDP-glucuronosyltransferase1 (UGT1A1 * 28),CYP2B6 * 6,methylene tetrahydrofolate reductase (MTHFR C677T),thiopurine S-methyltransferase (TPMT A719G),TPMT G238C,dihydropyrimidine dehydrogenase (DPYD * 2A14 + 1G ＞ A),DPYD * 9T85C.②Among examined specimens,100％ were of methotrexate sensitivity and66.7％ had temozolomide/carmustine/semustine sensitivity,52.2％ anthracycline/etoposide and fluorouracil sensitivity,52.1％ platinum sensitivity and 51.6％ vincristine sensitivity.While81.5％ had no sensitivity to irinotecan/topotecan,53.7％ non-sensitivity to cyclophosphamide.90.0％ weak toxicity to methotrexate,73.8％ weak toxicity to cyclophosphamide,64.3％ weak toxicity to fluorouracil and 55.6％ weak toxicity to irinotecan/etoposide.③Mesenchymal tumors had higher sensitivity to irinotecan/topotecan than non-mesenchymal tumors (P =0.000);NHL had higher sensitivity to anthracycline/etoposide than HL(P =0.000),HB had higher sensitivity to platinum(P =0.011)and vincristine (P =0.018)than NB.Based upon the above test results,therapeutic strategies for poor responders were promptly adjusted and then they achieved preliminary effect.Conclusion Genomic markers of drug sensitivity or toxicity are examined in children with malignant solid tumors.It provides rationales of formulating personalized chemotherapeutic regimens.