肺巨细胞癌高低转移株差异表达基因的鉴定  

Differentially expressed genes in human giant cell lung cancer lines with different metastatic potentials

作　　者：张金强[1] 王妍[1] 王涛[1] 杜芝燕[1] 徐元基[1] 陆应麟[1] 

机构地区：[1]军事医学科学院基础医学研究所病理生物学研究室,北京100850

出　　处：《中华肿瘤杂志》2004年第10期590-593,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金资助项目(30271452);国家重点基础研究发展规划项目(2002CB513105)

摘　　要：目的从两株同源但转移能力不同的人肺巨细胞癌细胞株中分离并鉴定差异表达基因。方法应用抑制消减杂交(SSH)技术,对来源相同、转移能力不同的人肺巨细胞癌细胞株PLA801C和PLA801D进行了两次实验。第1次SSH实验以PLA801C株为实验方,第2次实验以PLA801D株为实验方。将获得差异表达的片段点在氨基化的玻片上做成基因芯片,利用荧光标记的探针与芯片杂交,选取差异表达克隆,并利用RNA狭缝杂交或Northern杂交对若干片段进行验证。结果在低转移肺巨细胞癌细胞株中,高表达的序列有16条;在高转移肺巨细胞癌细胞株中,高表达的序列有79条。测序后经过同源性分析,大部分序列均与已知序列高度同源,主要编码以下几类蛋白(1)细胞因子及其受体相关蛋白;(2)激酶及其相关蛋白;(3)其他蛋白,包括酶类、热休克蛋白、受体蛋白、细胞骨架蛋白、线粒体蛋白和癌基因编码产物等;(4)一些未知功能的蛋白或是从核酸序列推出来的假想蛋白。结论HSP70、AXL受体酪氨酸激酶和1433ζ等多种已知基因表达情况的改变,可能影响肺癌的转移过程,此外还发现了一些可能的肿瘤转移相关新基因。Objective To screen genes differentially expressed in two human giant cell lung cancer lines of same origin but with different metastasis potentials. Methods Suppression subtractive hybridization (SSH) was done twice on two giant cell lung cancer lines, PLA 801C and PLA 801D (hereafter abbreviated as C and D), of same origin but with low (C) and high (D) metastatic potentials. In the first round, SSH C was used as tester and D as driver, while in the second round, the tester and driver were interchanged. The sequences acquired from both rounds of SSH were spotted on glass slides respectively and screened by hybridizing with two color fluorescence probes. Clones that had different expression levels on chips were also confirmed by RNA dot blot or Northern blot. Results There were 16 sequences with high expression in C as compared to those in D, and 79 sequences with high expression in D compared to those in C. After sequencing, most of them were found to be highly homologous to those encoding the following proteins: (1) cytokines and their receptors; (2) kinases and related proteins; (3) other proteins including enzymes, heat shock proteins, receptors, proteins of cell skeleton and mitochondria, products of oncogenes, etc; (4) some proteins deduced from gene sequences with yet unknown functions. Conclusion The alterations in expression of some known genes, including HSP70, AXL receptor tyrosine kinase and 14 3 3ζ, might have impact on metastasis of giant cell lung cancer. Whether some differentially expressed genes newly revealed are metastasis related needs further study. [

关 键 词：肺巨细胞癌 癌细胞株 差异表达基因 转移 高表达 蛋白 实验 激酶 玻片 序列 

分 类 号：R734.2[医药卫生—肿瘤]