论文部分内容阅读
目的 从两株人肺巨细胞癌细胞株中分离并鉴定人肺癌转移相关基因 ,为探讨肿瘤转移的分子机制奠定基础。方法 以细胞系PLA 80 1的两个具有不同转移潜力的亚克隆为模型 ,利用抑制消减杂交(suppressionsubtractivehybridization ,SSH)技术克隆差异片段 ,对得到的片段经基因芯片筛选后测序并与GenBank数据库进行同源性比较 ,然后用Northern杂交和RT PCR对其中的两条序列进行验证 ,并进行详细的生物信息学分析。结果 获得了 79条在高转移细胞中高表达而在低转移细胞中低表达的序列 ,在与这些序列同源的己知基因中包括两条功能未知的全长cDNA序列BC0 0 62 3 6和BC0 0 2 42 0 ,分别命名为MAG 1和MAG 2。MAG 1定位于 4q2 1,由 4个外显子组成 ,MAG 2定位于 2 q3 5 ,由 9个外显子组成 ,基因长度分别为8.5kb和 5 .2kb。二条序列均有开放阅读框架 (ORF) ,编码产物以α螺旋为主并含有部分片层结构和无规卷曲。结论 MAG 1和MAG 2在高低转移性肺巨细胞癌细胞株中的表达具有显著差异 ,二者的表达与高转移潜能具有相关性 ,提示MAG 1和MAG 2可能参与肿瘤转移过程。
Objective To isolate and identify human lung cancer metastasis-related genes from two human lung giant cell carcinoma cell lines and lay a foundation for exploring the molecular mechanism of tumor metastasis. Methods Two subclones of different cell lines PLA 80 1 with different metastatic potentials were used as a model to clone the different fragments by suppressionsubtractive hybridization (SSH). The obtained fragments were screened by gene chip and sequenced and compared with the GenBank database Then, the two sequences were verified by Northern blot and RT-PCR, and the detailed bioinformatics analysis was performed. As a result, 79 sequences highly expressed in highly metastatic cells and lowly expressed in low metastatic cells were obtained, including two unknown full-length cDNA sequences BC0 0 62 3 6 and BC0 0 among known genes homologous to these sequences 0 2 42 0, respectively named MAG 1 and MAG 2. MAG 1 is located at 4q21, consists of 4 exons, and MAG2 is located at 2q3 5, consisting of 9 exons with gene lengths of 8.5 kb and 5.2 kb, respectively. The two sequences are open reading frames (ORFs), the encoded products are mainly α-helix and contain some lamellar structure and random coil. Conclusion The expression of MAG 1 and MAG 2 in high and low metastatic human lung cancer cell lines is significantly different. The expression of MAG 1 and MAG 2 is correlated with the high metastatic potential, suggesting that MAG 1 and MAG 2 may play a role in tumor metastasis.