CYP2C19基因多态性分析及其与奥美拉唑疗效关系的临床研究
摘要
第一部分
50例吉林地区健康汉族人群和185例胃食管疾病患者CYP2C19基因多态性分析
目的：分析吉林地区健康汉族人群和胃食管疾病患者CYP2C19基因多态性。方法：50例无血缘关系的健康体检者参加本研究，其中男28例，女22例，年龄18～43岁，平均31.50±8.45岁。185例胃食管疾病患者。采用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）方法进行分析。结果：在50例健康汉族人中，弱代谢型共14例（14.00%），其中CYP2C19*2/CYP2C19*2型5例（10.00%），CYP2C19*2/ CYP2C19*3型2例（4.00%）；强代谢型共43例（86.00%），其中外显子5及外显子4均为CYP2C19*1/CYP2C19*1型者19例，占38.00%；CYP2C19*1/CYP2C19*2型20例，占40.00%；CYP2C19*1/ CYP2C19*3型4例，占8.00%。由此，CYP2C19*1的等位基因频率为0.62（95%CI:0.56-0.68），CYP2C19*2的为0.32（95%CI: 0.26-0.38），CYP2C19*3的为0.06（95%CI:0.02-0.10）。食管癌组中CYP2C19弱代谢者5例，与健康组相比有显著性差异(P<0.05；OR：4.39, 95%CI:1.08-17.77)。结论：我国吉林地区健康汉族人群中，弱代谢型频率约为14.00%，其中CYP2C19*2占多数，其等位基因频率约为CYP2C19*3的5倍。CYP2C19*3等位基因的频率尽管相对较低，但它几乎说明了中国人剩余的全部弱代谢型等位基因。CYP2C19弱代谢型与
食管癌的发生具有显著相关性，CYP2C19弱代谢型增加了发生食管癌的相对危险性。
关键词 CYP2C19; PCR-RFLP; 基因多态性
第二部分
奥美拉唑治疗反流性食管炎的疗效及其与CYP2C19基因多态性关系的临床研究
目的：探讨短期应用奥美拉唑治疗反流性食管炎的疗效及其与CYP2C19基因多态性的关系。方法：应用奥美拉唑20mg每日晨起空腹口服一次，持续4周，治疗56例经胃镜证实的反流性食管炎患者。记录治疗前和用药2周后、4周后时症状的变化，并于治疗结束时复查胃镜。受试者 CYP2C19的基因型及幽门螺杆菌感染情况分别采用聚合酶链式反应-限制性片段长度多态性（PCR－RFLP）及ELISA分析方法。结果：纯合子强代谢组、杂合子强代谢组和弱代谢组胃镜下治愈率分别为47.37%、69.23%和88.89%，纯合子强代谢组的治愈率显著低于弱代谢组（P<0.05）。性别差异并非影响反流性食管炎治愈率的主要因素。结论：CYP2C19基因多态性与奥美拉唑治疗反流性食管炎的疗效密切相关。CYP2C19基因分型检测是临床治疗酸相关性疾病中一个重要的工具。
Part I
Analysis of the CYP2C19 Polymorphism in 50 Chinese Han People and 185 Patients with Gastroesophageal Disease of JiLin Province
Objective: To assess the genotype pattern distribution of cytochrome P450 in Chinese Han population and patients with gastroesophageal disease of JiLin Province. Method: 50 unrelated healthy Chinese Han population subjects(28 males and 22 females, aged from 18～43) and 185 gastroesophageal disease patients were genotyped for CYP2C19 according to the polymerase chain reaction- restriction fragment length polymorphism technique(PCR-RFLP). Result: In 50 healthy people, 7 subjects (14.00%) were identified as poor metabolizers(PMs),consisting of 5 homozygous for CYP2C19*2/CYP2C19*2 and 2 heterozygous for CYP2C19*2/ CYP2C19*3,
there was no homozygous for CYP2C19*3/CYP2C19*3 being found. The frequency of extensive metabolizers(EMs) was 86.00% (43 of 50 subjects), consisting of 19(38.00%) homozygous for CYP2C19*1/ CYP2C19*1 in exon4 and exon5, 20(40.00%) heterozygous for CYP2C19*1/CYP2C19*2, 4(8.00%) heterozygous for CYP2C19*1/ CYP2C19*3. Thus allele frequencies for CYP2C19*1, CYP2C19*2 and CYP2C19*3 were 0.62 [95% confidence interval (CI):0.56-0.68], 0.32 (95% CI:0.26-0.38) and 0.06(95%CI: 0.02-0.10), respectively. 5 CYP2C19 poor metabolizers were found in esophageal cancer patients, which showed a very significant difference than that of healthy controls. Conclusion: The frequency of poor metabolizers in Chinese is 14.00%, whereas the frequency of extensive metabolizers is 86.00%, CYP2C19*2 is the majority in two mutations and frequency of the CYP2C19*2 allele is about 5 times than that of mutant allele CYP2C19*3.These results are consistent with the exhibited reports. CYP2C19 poor metabolizers increase the risk of esophageal cancer significantly.
Key words CYP2C19; genetic polymorphism; PCR-RFLP
Part II
Association between CYP2C19 Polymorphism and the Effects of Omeprazole on Reflux Esophagitis
Objective: To investigate whether the CYP2C19 genotype is related to the healing of erosive reflux esophagitis during treatment with omeprazole. Method: Fifty-six Chinese patients with reflux esophagitis were treated with a daily oral dose of
20 mg omeprazole for 4 weeks. The CYP2C19 genotype, Helicobacter pylori infection status were assessed before treatment. At 4 weeks, the healing of reflux esophagitis was evaluated endoscopically. Result: The healing rates were 47.37%, 69.23% and 88.89% at 4 weeks in homozygous extensive metabolizers, heterozygous extensive metabolizers and poor metabolizers, respectively. At 4 weeks, the healing rate of reflux esophagitis was significantly lower in homozygous extensive metabolizers than in poor metabolizers (P < 0.05). Sex had less influence than CYP2C19 polymorphism on the healing rate of reflux esophagitis. Conclusions: The therapeutic effect of omeprazole on reflux esophagitis is influenced by the CYP2C19 genotype status. Therefore, a test of CYP2C19 genotype may be useful for the medical treatment of reflux esophagitis with omeprazole.
50例吉林地区健康汉族人群和185例胃食管疾病患者CYP2C19基因多态性分析
目的：分析吉林地区健康汉族人群和胃食管疾病患者CYP2C19基因多态性。方法：50例无血缘关系的健康体检者参加本研究，其中男28例，女22例，年龄18～43岁，平均31.50±8.45岁。185例胃食管疾病患者。采用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）方法进行分析。结果：在50例健康汉族人中，弱代谢型共14例（14.00%），其中CYP2C19*2/CYP2C19*2型5例（10.00%），CYP2C19*2/ CYP2C19*3型2例（4.00%）；强代谢型共43例（86.00%），其中外显子5及外显子4均为CYP2C19*1/CYP2C19*1型者19例，占38.00%；CYP2C19*1/CYP2C19*2型20例，占40.00%；CYP2C19*1/ CYP2C19*3型4例，占8.00%。由此，CYP2C19*1的等位基因频率为0.62（95%CI:0.56-0.68），CYP2C19*2的为0.32（95%CI: 0.26-0.38），CYP2C19*3的为0.06（95%CI:0.02-0.10）。食管癌组中CYP2C19弱代谢者5例，与健康组相比有显著性差异(P<0.05；OR：4.39, 95%CI:1.08-17.77)。结论：我国吉林地区健康汉族人群中，弱代谢型频率约为14.00%，其中CYP2C19*2占多数，其等位基因频率约为CYP2C19*3的5倍。CYP2C19*3等位基因的频率尽管相对较低，但它几乎说明了中国人剩余的全部弱代谢型等位基因。CYP2C19弱代谢型与
食管癌的发生具有显著相关性，CYP2C19弱代谢型增加了发生食管癌的相对危险性。
关键词 CYP2C19; PCR-RFLP; 基因多态性
第二部分
奥美拉唑治疗反流性食管炎的疗效及其与CYP2C19基因多态性关系的临床研究
目的：探讨短期应用奥美拉唑治疗反流性食管炎的疗效及其与CYP2C19基因多态性的关系。方法：应用奥美拉唑20mg每日晨起空腹口服一次，持续4周，治疗56例经胃镜证实的反流性食管炎患者。记录治疗前和用药2周后、4周后时症状的变化，并于治疗结束时复查胃镜。受试者 CYP2C19的基因型及幽门螺杆菌感染情况分别采用聚合酶链式反应-限制性片段长度多态性（PCR－RFLP）及ELISA分析方法。结果：纯合子强代谢组、杂合子强代谢组和弱代谢组胃镜下治愈率分别为47.37%、69.23%和88.89%，纯合子强代谢组的治愈率显著低于弱代谢组（P<0.05）。性别差异并非影响反流性食管炎治愈率的主要因素。结论：CYP2C19基因多态性与奥美拉唑治疗反流性食管炎的疗效密切相关。CYP2C19基因分型检测是临床治疗酸相关性疾病中一个重要的工具。
Part I
Analysis of the CYP2C19 Polymorphism in 50 Chinese Han People and 185 Patients with Gastroesophageal Disease of JiLin Province
Objective: To assess the genotype pattern distribution of cytochrome P450 in Chinese Han population and patients with gastroesophageal disease of JiLin Province. Method: 50 unrelated healthy Chinese Han population subjects(28 males and 22 females, aged from 18～43) and 185 gastroesophageal disease patients were genotyped for CYP2C19 according to the polymerase chain reaction- restriction fragment length polymorphism technique(PCR-RFLP). Result: In 50 healthy people, 7 subjects (14.00%) were identified as poor metabolizers(PMs),consisting of 5 homozygous for CYP2C19*2/CYP2C19*2 and 2 heterozygous for CYP2C19*2/ CYP2C19*3,
there was no homozygous for CYP2C19*3/CYP2C19*3 being found. The frequency of extensive metabolizers(EMs) was 86.00% (43 of 50 subjects), consisting of 19(38.00%) homozygous for CYP2C19*1/ CYP2C19*1 in exon4 and exon5, 20(40.00%) heterozygous for CYP2C19*1/CYP2C19*2, 4(8.00%) heterozygous for CYP2C19*1/ CYP2C19*3. Thus allele frequencies for CYP2C19*1, CYP2C19*2 and CYP2C19*3 were 0.62 [95% confidence interval (CI):0.56-0.68], 0.32 (95% CI:0.26-0.38) and 0.06(95%CI: 0.02-0.10), respectively. 5 CYP2C19 poor metabolizers were found in esophageal cancer patients, which showed a very significant difference than that of healthy controls. Conclusion: The frequency of poor metabolizers in Chinese is 14.00%, whereas the frequency of extensive metabolizers is 86.00%, CYP2C19*2 is the majority in two mutations and frequency of the CYP2C19*2 allele is about 5 times than that of mutant allele CYP2C19*3.These results are consistent with the exhibited reports. CYP2C19 poor metabolizers increase the risk of esophageal cancer significantly.
Key words CYP2C19; genetic polymorphism; PCR-RFLP
Part II
Association between CYP2C19 Polymorphism and the Effects of Omeprazole on Reflux Esophagitis
Objective: To investigate whether the CYP2C19 genotype is related to the healing of erosive reflux esophagitis during treatment with omeprazole. Method: Fifty-six Chinese patients with reflux esophagitis were treated with a daily oral dose of
20 mg omeprazole for 4 weeks. The CYP2C19 genotype, Helicobacter pylori infection status were assessed before treatment. At 4 weeks, the healing of reflux esophagitis was evaluated endoscopically. Result: The healing rates were 47.37%, 69.23% and 88.89% at 4 weeks in homozygous extensive metabolizers, heterozygous extensive metabolizers and poor metabolizers, respectively. At 4 weeks, the healing rate of reflux esophagitis was significantly lower in homozygous extensive metabolizers than in poor metabolizers (P < 0.05). Sex had less influence than CYP2C19 polymorphism on the healing rate of reflux esophagitis. Conclusions: The therapeutic effect of omeprazole on reflux esophagitis is influenced by the CYP2C19 genotype status. Therefore, a test of CYP2C19 genotype may be useful for the medical treatment of reflux esophagitis with omeprazole.
引文
Zaphiropoulos PG. RNA molecules containing exons originating from different members of the cytochrome P450 2C gene subfamily (CYP2C) in human epidermis and liver. Nucleic Acids Res，1999；27:2585～2590
Obach RS, Zhang QY, Dunbar D, et al. Metabolic characterization of the major human small intestinal cytochrome p450s. Drug Metab Dispos，2001；29:347～352
Blaisdell J, Mohrenweiser H, Jackson J，et al. Identification and functional characterization of new potentially defective alleles of human CYP2C19. Pharmacogenetics，2002；12(9):703～711
Tsuneoka Y, Fukushima K, Matsuo Y, et al. Genotype analysis of the CYP2C19 gene in the Japanese population. Life Sci，1996；59:1711～1715
Brockm?ller J, Cascorbi I, Kerb R, et al. Combined analysis of inherited polymorphisms in arylamine N-acetyltransferase 2, glutathione S-transferases M1 and T1, microsomal epoxide hydrolase, and cytochrome P450 enzymes as modulators of bladder cancer risk. Cancer Res，1996；56:3915～3925
Roddam PL, Rollinson S, Kane E, et al. Poor metabolizers at the cytochrome P450 2D6 and 2C19 loci are at increased risk of developing adult acute leukemia. Pharmacogenetics， 2000；10:605～615
Xie HG, Kim RB, Stein CM, et al. Genetic polymorphism of (S)-mephenytoin 4'-hydroxylation in populations of African descent. Br J Clin Pharmacol，1999；48:402～408
8. Xie HG, Stein CM, Kim RB, et al. Allelic, genotypic and phenotypic distributions of S-mephenytoin 4'-hydroxylase (CYP2C19) in healthy Caucasian populations of European descent throughout the world. Pharmacogenetics，1999；9:539～549
9. Bertilsson L.Geographical/interracial differences in polymorphic drug oxidation. Clin Pharmacokinet，1995；29: 192～209
10.Goldstein JA, Ishizaki T, Chiba K, et al. Frequencies of the defective CYP2C19 alleles responsible for the mephenytoin poor olizer phenotype in various Oriental, Caucasian, Saudi Arabian and American Black populations. Pharmacogenetics，1997；7: 59～64
11.Roh HK, Dahl ML, Tybring G，et al. CYP2C19 genotype and phenotype determined by omeprazole in a Korean population. Pharmacogenetics，1996；6(6):547～55
12.de Morais SM, Wilkinson GR, Blaisdell J, et al The major genetic defect responsible for the polymorphism of S-mephenytoin metabolism in humans. J Biol Chem，1994；269: 15419～15422
13.de Morais SM, Wilkinson GR, Blaisdell J, et al. Identification of a new genetic defect responsible for the
polymorphism of S-mephenytoin metabolism in Japanese. Mol Pharmacol，1994；46:594～598
14.He N, Yan FX, Huang SL, et al.CYP2C19 genotype S-mephenytoin 4’-hytroxylation phenotype in Chinese Dai population. Eur J Clin Pharmacol，2002；58:15～18
15.张惠文，陈盛强.健康汉族人细胞色素P4502C19基因的检测 现代临床医学生物工程学杂志,2002；8（4）：261～262
16.唐莉、彭林、张松等.中国独龙族人群细胞色素P4502C19基因多态性的研究.中国药理学通报，2001；17（5）：556～558
17.张松、董兆文、唐莉等.中国苗族、布依族、土族和独龙族群体种细胞色素P4502C19基因座遗传多态性分析.中华医学遗传学杂志，2002；19（1）：52～54
18.段于峰、武占鹏、张松等.中国土族人群中细胞色素P4502C19基因多态性的研究.中国优生与遗传学杂志，2001；9(5):13～14
19.xiao ZS, Goldstein JA, Xie HG, et al. Differences in the incidence of the CYP2C19 polymorphism affecting the S-mephenytoin phenotype in Chinese Han and Bai populations and identification of a new rare CYP2C19 mutant alete. J Pharmacol Exp Ther, 1997；281(1):604～609
20.Kubota T, Chiba K, Ishizaki T. Genotyping of S-mephenytoin 4'-hydroxylation in an extended Japanese population. Clin Pharmacol Ther. 1996；60(6):661～666
21.Tassaneeyakul W, Tawalee A, Tassaneeyakul W, et al. Analysis of the CYP2C19 polymorphism in a North-eastern Thai population. Pharmacogenetics，2002；12(3): 221～225
22.Goldstein JA, Ishizaki T, Chiba K, et al. Frequencies of the defective CYP2C19 alleles responsible for the mephenytoin poor metabolizer phenotype in various Oriental, Caucasian, Saudi Arabian and American black populations. Pharmacogenetics，1997；7: 59–64
23.Tamminga WJ, Wemer J, Oosterhuis B，et al. The prevalence of CYP2D6 and CYP2C19 genotypes in a population of healthy Dutch volunteers. Eur J Clin Pharmacol，2001；57(10):717～722
24.Aynacioglu AS, Sachse C, Bozkurt A， et al. Low frequency of defective alleles of cytochrome P450 enzymes 2C19 and 2D6 in the Turkish population. Clin Pharmacol Ther，1999；66 (2): 185～192
25.Xiao ZS, Goldstein JA, Xie HG, et al. Differences in the incidence of the CYP2C19 polymorphism affecting the S-mephenytoin phenotype in Chinese Han and Bai populations and identification of a new rare CYP2C19 mutant allele. J Pharmacol Exp Ther，1997；281: 604–609
26.Xie HG, Huang SL, Xu ZH, et al. Evidence for the effect of gender on activity of (S)-mephenytoin 4'-hydroxylase (CYP2C19) in a Chinese population. Pharmacogenetics.1997；7(2):115～119
27.Xie HG, Stein CM, Kim RB, et al. Allelic, genotypic and phenotypic distributions of S-mephenytoin 4'-hydroxylase (CYP2C19) in healthy Caucasian populations of European descent throughout the world. Pharmacogenetics，1999；9(5):
539～549
28.Inaba T, Jorge LF, Arias TD. Mephenytoin hydroxylation in the Cuna Amerindians of Panama. Br J Clin Pharmacol， 1988；25: 75～79
29.Kaneko A, Laneko O, Taleo G, et al. High frequencies of CYP2C19 mutations and poor metabolism of proguanil in Vanuatu. Lancet，1997；349:921～922
Obach RS, Zhang QY, Dunbar D, et al. Metabolic characterization of the major human small intestinal cytochrome p450s. Drug Metab Dispos，2001；29:347～352
Blaisdell J, Mohrenweiser H, Jackson J，et al. Identification and functional characterization of new potentially defective alleles of human CYP2C19. Pharmacogenetics，2002；12(9):703～711
Tsuneoka Y, Fukushima K, Matsuo Y, et al. Genotype analysis of the CYP2C19 gene in the Japanese population. Life Sci，1996；59:1711～1715
Brockm?ller J, Cascorbi I, Kerb R, et al. Combined analysis of inherited polymorphisms in arylamine N-acetyltransferase 2, glutathione S-transferases M1 and T1, microsomal epoxide hydrolase, and cytochrome P450 enzymes as modulators of bladder cancer risk. Cancer Res，1996；56:3915～3925
Roddam PL, Rollinson S, Kane E, et al. Poor metabolizers at the cytochrome P450 2D6 and 2C19 loci are at increased risk of developing adult acute leukemia. Pharmacogenetics， 2000；10:605～615
Xie HG, Kim RB, Stein CM, et al. Genetic polymorphism of (S)-mephenytoin 4'-hydroxylation in populations of African descent. Br J Clin Pharmacol，1999；48:402～408
8. Xie HG, Stein CM, Kim RB, et al. Allelic, genotypic and phenotypic distributions of S-mephenytoin 4'-hydroxylase (CYP2C19) in healthy Caucasian populations of European descent throughout the world. Pharmacogenetics，1999；9:539～549
9. Bertilsson L.Geographical/interracial differences in polymorphic drug oxidation. Clin Pharmacokinet，1995；29: 192～209
10.Goldstein JA, Ishizaki T, Chiba K, et al. Frequencies of the defective CYP2C19 alleles responsible for the mephenytoin poor olizer phenotype in various Oriental, Caucasian, Saudi Arabian and American Black populations. Pharmacogenetics，1997；7: 59～64
11.Roh HK, Dahl ML, Tybring G，et al. CYP2C19 genotype and phenotype determined by omeprazole in a Korean population. Pharmacogenetics，1996；6(6):547～55
12.de Morais SM, Wilkinson GR, Blaisdell J, et al The major genetic defect responsible for the polymorphism of S-mephenytoin metabolism in humans. J Biol Chem，1994；269: 15419～15422
13.de Morais SM, Wilkinson GR, Blaisdell J, et al. Identification of a new genetic defect responsible for the
polymorphism of S-mephenytoin metabolism in Japanese. Mol Pharmacol，1994；46:594～598
14.He N, Yan FX, Huang SL, et al.CYP2C19 genotype S-mephenytoin 4’-hytroxylation phenotype in Chinese Dai population. Eur J Clin Pharmacol，2002；58:15～18
15.张惠文，陈盛强.健康汉族人细胞色素P4502C19基因的检测 现代临床医学生物工程学杂志,2002；8（4）：261～262
16.唐莉、彭林、张松等.中国独龙族人群细胞色素P4502C19基因多态性的研究.中国药理学通报，2001；17（5）：556～558
17.张松、董兆文、唐莉等.中国苗族、布依族、土族和独龙族群体种细胞色素P4502C19基因座遗传多态性分析.中华医学遗传学杂志，2002；19（1）：52～54
18.段于峰、武占鹏、张松等.中国土族人群中细胞色素P4502C19基因多态性的研究.中国优生与遗传学杂志，2001；9(5):13～14
19.xiao ZS, Goldstein JA, Xie HG, et al. Differences in the incidence of the CYP2C19 polymorphism affecting the S-mephenytoin phenotype in Chinese Han and Bai populations and identification of a new rare CYP2C19 mutant alete. J Pharmacol Exp Ther, 1997；281(1):604～609
20.Kubota T, Chiba K, Ishizaki T. Genotyping of S-mephenytoin 4'-hydroxylation in an extended Japanese population. Clin Pharmacol Ther. 1996；60(6):661～666
21.Tassaneeyakul W, Tawalee A, Tassaneeyakul W, et al. Analysis of the CYP2C19 polymorphism in a North-eastern Thai population. Pharmacogenetics，2002；12(3): 221～225
22.Goldstein JA, Ishizaki T, Chiba K, et al. Frequencies of the defective CYP2C19 alleles responsible for the mephenytoin poor metabolizer phenotype in various Oriental, Caucasian, Saudi Arabian and American black populations. Pharmacogenetics，1997；7: 59–64
23.Tamminga WJ, Wemer J, Oosterhuis B，et al. The prevalence of CYP2D6 and CYP2C19 genotypes in a population of healthy Dutch volunteers. Eur J Clin Pharmacol，2001；57(10):717～722
24.Aynacioglu AS, Sachse C, Bozkurt A， et al. Low frequency of defective alleles of cytochrome P450 enzymes 2C19 and 2D6 in the Turkish population. Clin Pharmacol Ther，1999；66 (2): 185～192
25.Xiao ZS, Goldstein JA, Xie HG, et al. Differences in the incidence of the CYP2C19 polymorphism affecting the S-mephenytoin phenotype in Chinese Han and Bai populations and identification of a new rare CYP2C19 mutant allele. J Pharmacol Exp Ther，1997；281: 604–609
26.Xie HG, Huang SL, Xu ZH, et al. Evidence for the effect of gender on activity of (S)-mephenytoin 4'-hydroxylase (CYP2C19) in a Chinese population. Pharmacogenetics.1997；7(2):115～119
27.Xie HG, Stein CM, Kim RB, et al. Allelic, genotypic and phenotypic distributions of S-mephenytoin 4'-hydroxylase (CYP2C19) in healthy Caucasian populations of European descent throughout the world. Pharmacogenetics，1999；9(5):
539～549
28.Inaba T, Jorge LF, Arias TD. Mephenytoin hydroxylation in the Cuna Amerindians of Panama. Br J Clin Pharmacol， 1988；25: 75～79
29.Kaneko A, Laneko O, Taleo G, et al. High frequencies of CYP2C19 mutations and poor metabolism of proguanil in Vanuatu. Lancet，1997；349:921～922