塞替派诱发人支气管上皮细胞恶性转化过程中的基因突变(二)  

Detection of mutations of p53,p16 and Ki-ras genes in the process of human bronchial epithelial cells malignant transformation induced by thiotepa.

作　　者：袁素波[1] 夏英[2] 叶常青[3] 廖明阳[1] 王治乔[1] 杨梅英[3] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850 [2]卫生部工业卫生实验所 [3]军事医学科学院放射医学研究所

出　　处：《卫生毒理学杂志》2001年第4期211-215,共5页Journal of Health Toxicology

基　　金：北京市科委实验动物专项基金资助 ( 95 40 2 45 0 0 )

摘　　要：目的　观察抗癌药物塞替派诱发永生化人支气管上皮细胞恶性转化过程中相关基因的突变并分析其意义。方法　以塞替派作为致癌原诱导永生化人支气管上皮细胞 (BEAS 2B) ,获得转化细胞 (BEAS TE)和克隆化多倍体癌前转化细胞 (BEAS STE)。采用PCR SSCP法检测上述 3种细胞p5 3、p16和Ki ras 3种基因是否出现点突变 ,进一步测序确定其突变情况。结果　SSCP结果阳性的有BEAS TE细胞p5 3第 7外显子 ,BEAS STE细胞p5 3第8外显子以及这二种细胞的p16基因第 1外显子 ;Ki ras基因第 1外显子的结果仅为可疑阳性。测序证明 ,p5 3、Ki ras基因存在多位点的碱基突变 ,而p16基因仅为单位点的碱基突变。结论　塞替派可诱发人支气管上皮细胞p5 3、Ki ras多位点的碱基突变和p16的单位点突变。分析前者为塞替派诱导细胞转化过程中发生的重要分子事件 。Objective To observe the mutation of related genes during the process of malignant transformation induced by a chemotherapeutic agent thiotepa in the immortalized human bronchial epithelial cell line (BEAS 2B).Methods The thiotepa was adopted as a carcinogen which induced the BEAS 2B to be malignantly transformed into transformed cells (BEAS TE)and coloned multi ploid precarcino transformed cells (BEAS STE). PCR SSCP was used to detect the point mutation of p53,p16 and Ki ras in three cells above, the mutation exons were analyzed by gene sequence.Results There were the positive results of PCR SSCP in the exon 7 of p53 of BEAS TE, exon 8 of p53 of BEAS STE, and exon 1 of p16 of both exposed cells, but there were only the suspicious positive results showed in the exon 1 of both exposed cells. Mutil sites base mutation was found in the genes of p53 and Ki ras , but only single site one in the gene of p16 .Conclusion Multi sites base mutation of gene p53, Ki ras and single site base mutation of gene p16 in the BEAS 2B could be induced by thiotepa, it seems that the former was the key molecular events,While the latter was the secondary one in the cell transformation process.

关 键 词：塞替派 永生化人支管上皮细胞 BEAS-2B p53 P16 KI-RAS基因 突变 PCR-SSCR 序列分析 

分 类 号：R99[医药卫生—毒理学] R979.1[医药卫生—药学]