中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2011年
5期
393-398
,共6页
张爱华%潘雪莉%夏玉洁%肖骞%黄晓欣
張愛華%潘雪莉%夏玉潔%肖鶱%黃曉訢
장애화%반설리%하옥길%초건%황효흔
砷中毒%煤%基因,p53%DNA甲基化%突变
砷中毒%煤%基因,p53%DNA甲基化%突變
신중독%매%기인,p53%DNA갑기화%돌변
Arsenic poisoning%Coal%Genes,p53%DNA methylation%Mutation
目的 探讨燃煤砷污染对人体p53基因甲基化(启动子区及第5外显子)和突变(第5外显子)的影响,分析其与燃煤型砷中毒的关系.方法 在贵州省兴仁县燃煤型砷中毒病区选择112例砷中毒患者,根据病情将其分为轻度中毒组(38例)、中度中毒组(43例)和重度中毒组(31例);根据有皮肤病理学诊断患者(43例)的病理结果,将其分为非癌变组(24例)和癌变组(19例).选择条件相似的非砷暴露村90名居民作为对照组.在知情同意原则下,采集上述观察对象的外周血,应用限制性内切酶-PCR法检测p53基因启动子区及第5外显子甲基化情况,应用PCR-单链构象多态性技术及PCR产物克隆测序技术检测p53基因第5外显子突变情况.结果 p53基因启动子区甲基化阳性率在轻、中、重度组分别为13.16%(5/38)、27.91%(12/43)、45.16%(14/31),与对照组[1.11%(1/90)]比较差异均有统计学意义(χ2值分别为8.679、23.690、41.199,P值均＜0.017);在非癌变组和癌变组分别为25.00%(6/24)和63.16%(12/19),与对照组[1.11%(1/90)]比较差异均有统计学意义(χ2值分别为18.762、57.497,P值均＜0.025).p53基因第5外显子甲基化阳性率在轻、中、重度组分别为55.26%(21/38)、51.16%(22/43)、48.39%(15/31),与对照组[88.88%(80/90)]比较差异有统计学意义(χ2值分别为18.151、23.168、22.420,P值均＜0.017);在非癌变组和癌变组分别为54.17%(13/24)和42.11%(8/19),与对照组[88.88%(80/90)]比较差异有统计学意义(χ2值分别为15.201、22.075,P值均＜0.025).p53基因第5外显子突变率在轻、中、重度组分别为5.26%(2/38)、16.28%(7/43)、25.81%(8/31),中、重度组与对照组(0.00%)比较差异均有统计学意义(χ2值分别为15.465、24.870,P值均＜0.017);在非癌变组和癌变组分别为16.67%(4/24)、31.58%(6/19),与对照组(0.00%)比较差异均有统计学意义(χ2值分别为15.545、30.077,P值均＜0.025).启动子区高甲基化与第5外显子突变相关(列联系数为0.294,P＜0.05);第5外显子低甲基化与第5外显子突变相关(列联系数为0.410,P＜0.05).结论 燃煤砷污染可致人体p53基因启动子区高甲基化、第5外显子低甲基化和突变,上述甲基化改变均与p53基因突变相关联;p53基因甲基化改变可能是砷致p53基因突变以及砷致病或致癌的重要原因之一.
目的 探討燃煤砷汙染對人體p53基因甲基化(啟動子區及第5外顯子)和突變(第5外顯子)的影響,分析其與燃煤型砷中毒的關繫.方法 在貴州省興仁縣燃煤型砷中毒病區選擇112例砷中毒患者,根據病情將其分為輕度中毒組(38例)、中度中毒組(43例)和重度中毒組(31例);根據有皮膚病理學診斷患者(43例)的病理結果,將其分為非癌變組(24例)和癌變組(19例).選擇條件相似的非砷暴露村90名居民作為對照組.在知情同意原則下,採集上述觀察對象的外週血,應用限製性內切酶-PCR法檢測p53基因啟動子區及第5外顯子甲基化情況,應用PCR-單鏈構象多態性技術及PCR產物剋隆測序技術檢測p53基因第5外顯子突變情況.結果 p53基因啟動子區甲基化暘性率在輕、中、重度組分彆為13.16%(5/38)、27.91%(12/43)、45.16%(14/31),與對照組[1.11%(1/90)]比較差異均有統計學意義(χ2值分彆為8.679、23.690、41.199,P值均＜0.017);在非癌變組和癌變組分彆為25.00%(6/24)和63.16%(12/19),與對照組[1.11%(1/90)]比較差異均有統計學意義(χ2值分彆為18.762、57.497,P值均＜0.025).p53基因第5外顯子甲基化暘性率在輕、中、重度組分彆為55.26%(21/38)、51.16%(22/43)、48.39%(15/31),與對照組[88.88%(80/90)]比較差異有統計學意義(χ2值分彆為18.151、23.168、22.420,P值均＜0.017);在非癌變組和癌變組分彆為54.17%(13/24)和42.11%(8/19),與對照組[88.88%(80/90)]比較差異有統計學意義(χ2值分彆為15.201、22.075,P值均＜0.025).p53基因第5外顯子突變率在輕、中、重度組分彆為5.26%(2/38)、16.28%(7/43)、25.81%(8/31),中、重度組與對照組(0.00%)比較差異均有統計學意義(χ2值分彆為15.465、24.870,P值均＜0.017);在非癌變組和癌變組分彆為16.67%(4/24)、31.58%(6/19),與對照組(0.00%)比較差異均有統計學意義(χ2值分彆為15.545、30.077,P值均＜0.025).啟動子區高甲基化與第5外顯子突變相關(列聯繫數為0.294,P＜0.05);第5外顯子低甲基化與第5外顯子突變相關(列聯繫數為0.410,P＜0.05).結論 燃煤砷汙染可緻人體p53基因啟動子區高甲基化、第5外顯子低甲基化和突變,上述甲基化改變均與p53基因突變相關聯;p53基因甲基化改變可能是砷緻p53基因突變以及砷緻病或緻癌的重要原因之一.
목적 탐토연매신오염대인체p53기인갑기화(계동자구급제5외현자)화돌변(제5외현자)적영향,분석기여연매형신중독적관계.방법 재귀주성흥인현연매형신중독병구선택112례신중독환자,근거병정장기분위경도중독조(38례)、중도중독조(43례)화중도중독조(31례);근거유피부병이학진단환자(43례)적병리결과,장기분위비암변조(24례)화암변조(19례).선택조건상사적비신폭로촌90명거민작위대조조.재지정동의원칙하,채집상술관찰대상적외주혈,응용한제성내절매-PCR법검측p53기인계동자구급제5외현자갑기화정황,응용PCR-단련구상다태성기술급PCR산물극륭측서기술검측p53기인제5외현자돌변정황.결과 p53기인계동자구갑기화양성솔재경、중、중도조분별위13.16%(5/38)、27.91%(12/43)、45.16%(14/31),여대조조[1.11%(1/90)]비교차이균유통계학의의(χ2치분별위8.679、23.690、41.199,P치균＜0.017);재비암변조화암변조분별위25.00%(6/24)화63.16%(12/19),여대조조[1.11%(1/90)]비교차이균유통계학의의(χ2치분별위18.762、57.497,P치균＜0.025).p53기인제5외현자갑기화양성솔재경、중、중도조분별위55.26%(21/38)、51.16%(22/43)、48.39%(15/31),여대조조[88.88%(80/90)]비교차이유통계학의의(χ2치분별위18.151、23.168、22.420,P치균＜0.017);재비암변조화암변조분별위54.17%(13/24)화42.11%(8/19),여대조조[88.88%(80/90)]비교차이유통계학의의(χ2치분별위15.201、22.075,P치균＜0.025).p53기인제5외현자돌변솔재경、중、중도조분별위5.26%(2/38)、16.28%(7/43)、25.81%(8/31),중、중도조여대조조(0.00%)비교차이균유통계학의의(χ2치분별위15.465、24.870,P치균＜0.017);재비암변조화암변조분별위16.67%(4/24)、31.58%(6/19),여대조조(0.00%)비교차이균유통계학의의(χ2치분별위15.545、30.077,P치균＜0.025).계동자구고갑기화여제5외현자돌변상관(렬련계수위0.294,P＜0.05);제5외현자저갑기화여제5외현자돌변상관(렬련계수위0.410,P＜0.05).결론 연매신오염가치인체p53기인계동자구고갑기화、제5외현자저갑기화화돌변,상술갑기화개변균여p53기인돌변상관련;p53기인갑기화개변가능시신치p53기인돌변이급신치병혹치암적중요원인지일.
Objective To explore the influence of arsenic pollution caused by coal-burning on methylation(promoter and exon 5 ) and mutation (exon 5 ) of human p53 gene, and to analyze the relationship between methylation, mutation and arsenism. Methods According to the diagnostic criteria of endemic arsenism, 112 patients with arsenism (including 38 mild cases,43 moderate cases and 31 severe cases) were selected in the areas with endemic arsenism from Xingren, Guizhou province. Among the subjects ,43 cases were diagnosed by dermatopathological methods, and they were divided into non-cancerous group (24 cases) and cancerous group ( 19 cases ). 90 controls were selected from the non-arsenic polluted areas. Under the principle of informed consent, blood samples were collected from individuals. The methylation of p53 gene in promoter region and exon 5 were detected by extinction enzyme-PCR,the mutation of p53 gene (exon 5 ) was detected by PCR-SSCP,PCR products cloning and sequencing technology. Results The positive rates of methylation of p53 gene in promoter region were 13. 16% ( 5/38 ), 27.91% ( 12/43 )and 45. 16% (14/31) respectively among mild, moderate and severe arsenism group, which were obviously higher than the rates in the control group (1.11% (1/90), χ2 values were 8.679,23.690, 41. 199,respectively,both P values ＜ 0. 017 ). The positive rates of methylation of p53 gene were 25.00% (6/24)and 63. 16% (12/19)in non-cancerous and cancerous group respectively, which were obviously higher than those in the control group ( 1. 11% (1/90) ,χ2 values were 18. 762,57. 497,respectively,both P values ＜0. 025 ). The positive rates of methylation of p53 gene ( exon 5 ) were 55.26% ( 21/38 ), 51. 16% ( 22/43 )and 48. 39% (15/31)respectively among mild, moderate and severe arsenism group,which were obviously lower than the rates in the control group ( 88. 88% ( 80/90 ), χ2 values were 18. 151 , 23. 168,22. 420,respectively, both P values ＜ 0. 017 ). The positive rates of methylation of p53 gene (exon 5 ) were 54. 17%(13/24) and 42. 11% (8/19) in non-cancerous and cancerous group respectively, which were obviously lower than those in the control group ( 88. 88% (80/90), χ2 values were 15. 201,22. 075, respectively, both P values ＜ 0. 025 ). The mutation rates of p53 gene ( exon 5 ) were respectively 5. 26% ( 2/38 ), 16. 28%(7/43) and 25.81% (8/31 )among mild, moderate and severe arsenism group; while the results in moderate and severe arsenism group were obviously higher than in the control group (0. 00% ,χ2 values were 15.465,24. 870,respectively,both P values ＜ 0. 017). The positive rate of mutation of p53 gene ( exon 5 ) were respectively 16. 67% (4/24) and 31.58% ( 6/19 ) in non-cancerous and cancerous group, which were obviously higher than it in the control group (0. 00%, χ2 values were 15. 545,30. 077, both P values ＜0. 025). The hypermethylation of p53 gene in promoter region was related with the mutation of p53 gene ( exon 5) ( coefficient of association was 0. 294, P value ＜ 0. 05 ); and the hypomethylation of p53 gene (exon 5 ) was related with the its mutation ( coefficient of association was 0. 410, P value ＜ 0. 05 ).Conclusion Arsenic pollution caused by coal-burning can cause the hypermethylation of p53 gene in promoter region, hypomethylation and mutation of p53 gene ( exon 5 ), and the changes of methylation of p53 gene are related with its mutation and might be one of the important etiological factors of arsenic pathogenicity or carcinogenesis.
