人肺癌高低转移细胞株差异表达基因的筛选

目的 筛选高低转移能力人肺癌细胞株PG-BE1和PG-LH7间差异表达基因.方法 进行两次抑制性消减杂交,第一次以PG-BE1细胞株为实验方、PG-LH7株为驱动方构建正向消减文库;第二次以PG-LH7株为实验方、PG-BE1为驱动方构建反向消减文库.筛选出来的阳性克隆进行测序,并在Gene Bank数据库中进行同源性比较.结果 2个消减文库共得到122个阳性克隆,随机选取10个克隆测序并进行同源性分析,发现其中9条序列来自于已知基因,另外1条序列为新的基因序列标签.结论 成功构建了高低转移力人肺癌细胞株的双向消减杂交文库.

Screen of differently expressed genes in human lung cancer cell lines with different metastasis potentials

Objective To screen genes differentially expressed in two human lung cancer cell lines PG-BE1 and PG-LH7 with different metastasis potentials. Methods Suppression subtractive hybridization was done twice on human lung cancer cell lines with high potential of metastasis PG-BE1 and its synogenetic cell line PG-LH7 with low metastasis potential. The first subtraction used PG-LH7 as tester and PG-BE1 as driver and constructed the forward subtractive library, while the second one interchanged the tester and driver and constructed the reverse subtractive library. The screened clones of both libraries were sequenced and Gene Bank homology search was performed. Results Two subtractive libraries containing 122 positive clones were constructed. Analysis of 10 sequenced clones randomly picked from the libraries showed that 1 differentially expressed gene fragments were identified as new EST with unknown functions. Conclusion The two subtractive libraries of human lung cancer cell lines with different metastasis potentials were constructed successfully.