摘要
目的 观察细胞凋亡相关基因的表达。方法 天花粉蛋白诱导Ｕ９３７细胞凋亡后，用锚定引物和随机引物进行差异显示逆转录ＰＣＲ反应，回收差异条带，进行ＲＮＡ点杂交。用细胞凋亡时高表达的差异片段筛选胎肝ｃＤＮＡ文库，随后测序，行Ｎｏｒｔｈｅｒｎ杂交。结果 用细胞凋亡时高表达的差异片段筛选胎肝ｃＤＮＡ文库，获得一个１－４ｋｂ的ｃＤＮＡ克隆，该ｃＤＮＡ序列中部分碱基与人Ｂｒｕｔｏｎ○ｓ酪氨酸激酶高度同源。用１－４ｋｂｃＤＮＡ进行Ｎｏｒｔｈｅｒｎ杂交证实该基因在细胞中有３个转录本存在，其中０－７ｋｂ的ｍＲＮＡ在凋亡细胞中高表达。同时观察到Ｄｒｇ１基因高表达。结论 用差异显示逆转录ＰＣＲ方法结合ｃＤＮＡ文库筛选，寻找到１－４ｋｂ的ｃＤＮＡ片段，部分序列与Ｂｒｕｔｏｎｓ酪氨酸激酶高度同源，是与细胞凋亡有关的新基因的部分片段。凋亡相关基因Ｄｒｇ１在天花粉蛋白诱导Ｕ９３７细胞凋亡时高表达。
AIM To search for the genes related to apoptosis. METHODS The different mRNA expression patterns of U937 cells treated with or without trichosanthin were compared with differential display RT PCR method. The cDNA segments, which were highly expressed in the apoptosis cells, were used as probes to screen the human fetal liver cDNA library and to hybridize with the total RNA from U937 in the two groups with dot blot analysis. RESULTS A 1 4 kb cDNA segment is the partial sequence of a novel gene related to apoptosis ,the result of Northern blot analysis showed three bands in the apoptosis cells. Partial sequence of the 1 4 kb cDNA was homology to Brutons tyrosine kinase. The Drg 1 gene was highly expressed in apoptosis of U937 cells. CONCLUSIONS The partial cDNA of a new gene related to apoptosis and the high expression of Drg 1 in U937 apoptosis induced by trichosanthin were found.
出处
《中国药理学通报》
CAS
CSCD
北大核心
1999年第6期516-518,共3页
Chinese Pharmacological Bulletin
基金
上海市医学领先专业专项基金
国家杰出青年科学基金!No 39725029
关键词
天花粉蛋白
细胞凋亡
基因表达
U937 cell
apoptosis
differential display RTPCR
