镉和镍恶性转化16HBE上调基因eIF3 p36的克隆与序列  

Cloning and sequencing of gene fragments of translation factor eIF3 p36 in malignant transformed human bronchial epithelial cells induced by crystalline nickel sulfide and cadmium chloride

作　　者：李敏[1] 雷毅雄[1] 魏莲[1] 邹晓妮[1] 

机构地区：[1]广州医学院公共卫生与全科医学学院,广东广州510182

出　　处：《中国热带医学》2007年第4期497-498,514,共3页China Tropical Medicine

基　　金：国家自然科学基金资助项目(30371195);广东省自然科学基金资助项目(06022672;031756);广州市科技攻关重点资助项目(2003Z2-E0191/E0192);广州市市属高校科技计划项目(编号:1002);广州医学院自然科学基金重点项目(编号:2006ZR004)

摘　　要：目的分析镉和镍恶性转化16HBE(人支气管上皮细胞)成瘤细胞株中翻译起始因子eIF3 p36基因序列的变化情况,探讨镉、镍重金属的致癌分子机制。方法取正常对照16HBE细胞、氯化镉(CdCl2)和结晶型硫化镍(NiS)恶性转化的16HBE细胞,用有限稀释法分别建立单细胞克隆株,通过逆转录聚合酶链反应(RT-PCR)获得目的基因翻译起始因子eIF3p36的cDNA,将目的基因产物连接到T载体中,转化、扩增重组质粒。提取大肠杆菌中的重组质粒进行DNA测序。将三组DNA测序所得到的基因序列进行对比分析。结果镉、镍转化组各细胞株(各10株)翻译起始因子的eIF3 p36cDNA测序结果与正常对照细胞(10株)比较未发现基因的突变位点;所有镉、镍转化细胞和正常对照细胞株eIF3 p36cDNA序列与Genbank数据库进行同源性分析,相似性比值均为100%。结论实验中镉、镍恶性转化16HBE细胞中翻译起始因子eIF3p36上调未见基因序列改变,提示在镉、镍致癌中可能涉及到表遗传机制。Objective To explore the molecular mechanism potentially responsible for carcinogenesis due to cadmium and nickel by cloning and sequencing the gene fragment of translation factor cIF3 in malignant transformed human bronchial epithelial cells in- duced by crystalline nickel and cadmium chloride, Methods Three lines of single - cell clones were cloned. Total RNA was isolated from nontransformed cells and tumorigenic cells using the TRIzol kit, The translation factor＇s （TIFp36） eDNA was obtained using RT- PCR methods. The PCR products were purified and then cloned into PMD 18 - T vector followed by sequencing. Resuits Compared with those of control 16HBE cells, no change in the sequence of eIF3 p36 cDNA in tumorigenic cells induced by Cadmium chloride and Nickel sulfide was detected, Conclusion The eIF3 p36 CDNA region might not be a target sequence in carcinogenesis induced by heavy metal. The carcinogenesis may be related to the unheredily mechanism.

关 键 词：人支气管上皮细胞 氯化镉 硫化镍 翻译起始因子 DNA测序 

分 类 号：R994.3[医药卫生—毒理学]