PCR-SSCP及克隆测序法研究燃煤型砷中毒患者p53基因突变与皮肤癌的关系

A Study on the Relationship between the Mutation of p53 Gene and Skin Cancer in Patients with Arsenism Caused by Coal-burning Using PCR-SSCP,Cloning and Sequencing

[目的 ]分析燃煤型砷中毒患者外周血中p5 3基因突变情况。 [方法 ]采用聚合酶链反应 单链构象多态性(PCR -SSCP)技术对 60例燃煤型砷中毒患者外周血中p5 3基因 5～ 8外显子的突变情况进行初筛 ,并进一步克隆测序 ,以确定突变位点及类型。 [结果 ]癌变组p5 3基因突变率为 3 0 % ( 3 /10 ) ;癌前组为 16.67% ( 2 /12 ) ;一般病变组未发现突变 ( 0 /3 8)。对筛查出来的 5例突变阳性者进行克隆测序 ,突变均为G :C→A :T的转换 ,分别位于密码子 14 3、14 6及 15 1。[结论 ]燃煤型砷中毒患者外周血中p5 3基因突变的检出 。

To study the mutation of p53 gene in peripheral blood white cells of patients with arsenism caused by coal-burning pollution. PCR- SSCP was performed to detect mutation of p53 gene exon 5～8 in peripheral blood white cells of patients with arsenism caused by coal-burning pollution,then,PCR product cloning,sequencing were performed to find the types and sites of mutation. p53 gene mutations were found in 30% of cases in carcinoma group(3/10),16.67% of cases with pre-carcinoma group(2/12),there was no p53 gene mutation found in 38 cases in general pathological group. The results of cloning and sequencing indicated that mutation types were G:C→A:T transition,located at codons 143,146 and 151. [Conclusion] Detection of p53 mutation in peripheral blood white cells is very important with theoretical values to reveal the mechanism of arsenic carcinogenicity.