Resarch of novel tumor metastasis-related genes for lung canncer

原发肿瘤的恶性细胞侵袭和随后的转移严重影响癌症患者的死亡率，但涉及的遗传和生物学机制仍然知之甚少。转移是一系列复杂的事件，其中一些基因起刺激作用，而另一些基因则抑制这一过程；据推测，几个这样的基因之间的不平衡导致了转移。我们使用了一种结合减法抑制杂交（SSH）和高通量差异筛选的系统来寻找转移相关基因。我们实验室从人肺腺癌中建立了HAL细胞系，其中HAL-8亚系是裸鼠肌内注射的转移细胞系，而HAL-24细胞则不是。采用SSH技术提取区分转移性HAL-8和非转移性HAL-24细胞的RNA。将减法的cDNA片段转染到组份细胞中，从组份细胞的转录本中提取cDNA片段进行序列分析。将PCR获得的序列与基因组数据库进行比较，得到了几个代表性基因的分离DNA和与滋养层弱连锁的新基因，后者是分析HAL细胞的cDNA文库。

Invasion of malignant cells from primary tumors and subsequent metastasis critically influence mortality among cancer patients, but the genetic and biological mechanisms involved are still poorly understood. Metastasis is a complex series of events in which some genes function as stimulatory factors and others inhibit the process ; presumably, an imbalance among several such genes induces metastasis.We have used a system that combines the subtractive suppression hybridization (SSH) with high throughput differential screening for search of metastasis related genes. The cell lines HAL were establishment from human lung adenocarcinoma in our laboratory, the sublines HAL-8 was metastatic cell line to inject intramuscularly in nude mice while HAL-24 cells do not. SSH technique was used to subtract the RNA distinguishing metastatic HAL-8 and non-metastatic HAL-24 cells.The cDNA fragments of subtract were trasfected to the compitents cells and recovered from the transcripts in compitents cells were analyzed by sequences. Comparison of the sequences obtained by PCR with genome databases provided that the isolated DNA of several represented genes and novel genes for weakly linked to trophoblast, the latter is analyzing cDNA library of HAL cells.