高淋巴系统转移能力小鼠肝癌细胞株差异表达基因的筛选

Screening of differentially expressed genes in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis

目的 筛选高淋巴系统转移能力小鼠肝癌细胞株差异性表达基因。方法 应用抑制性消减杂交 (SSH)技术 ,构建高淋巴系统转移能力小鼠肝癌细胞株Hca F及其同源低转移细胞系Hca P的cDNA消减杂交文库 ,筛选阳性克隆进行测序 ,并在GenBank数据库中进行同源性比较。结果 成功克隆 10个差异基因片段 ,其中 2个为新基因。结论 SSH技术是克隆差异表达基因及发现新基因的有效方法。

Objective To screen genes differentially expressed in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis. Methods A subtracted cDNA library of mouse hepatocarcinoma cell line with high potential of lymphatic metastasis Hca F and its synogenetic cell line Hca P with low metastatic potential was constructed by suppression subtractive hybridization(SSH) method. The screened clones of the subtracted library were sequenced and GenBank homology search was performed. Results Ten differentially expressed cDNA fragments of Hca F with high potential of lymphatic spreading were obtained, two of which were newly identified ones. Conclusion SSH is a useful technique to detect genes of differential expression and an effective method to clone novel genes.