摘要
目的：建立ＤＮＡ引物酶抑制剂的筛选方法，并应用于大量筛选，找出对ＤＮＡ引物酶有抑制作用的抗癌药物或先导化合物。方法：以小鼠艾氏腹水癌细胞破碎液为材料，过ＤＥ－５２和Ｐ－１１层析柱，提取分离ＤＮＡ引物酶，用大肠杆菌大片段ＤＮＡ多聚酶Ⅰ延长引物法检测ＤＮＡ引物酶的活性。并在此反应液中加入化合物检测对ＤＮＡ引物酶活性的抑制作用，以激活的小牛胸腺ＤＮＡ为模板排除化合物对大片断ＤＮＡ多聚酶Ⅰ的抑制作用。结果：部分纯化的ＤＮＡ引物酶比活性为８７５３Ｕ／ｍｇ，酶的总获得率为２２１％。在１００μｇ／ｍｌ浓度下，氯霉素和鞘氨醇对ＤＮＡ引物酶活性抑制率分另为７１２％±３７％和８３０％±５４％。结论：该方法已经建立，大量筛选发现非抗癌药物的氯霉素和鞘氨醇两种化合物对ＤＮＡ引物酶活性有抑制作用。临床常用的抗癌药物未发现对此酶活性有抑制作用。
Objective: To establish
the screening method of DNA primase inhibitors.The leading compouds of inhibiting DNA
primase activity were found through this method. Methods: DNA primase was extracted from
mouse EAC cells, then was partially purified and identified.Its activity was assayed using
indirect method, namely polydT,ATP,3HdATP as substrate,and 02u E.coli.DNA polymerase
Klenow fragment was added to the reaction mixture.The inhibitory effects of the drugs on DNA
primase activity were assayed. Results: The effect of drugs on DNA primase activity were
studied.Under the concentration of 100 g/ml ,inhibitory rate of chloramphenicol and sphingosine
were 705%27%and 830%54%.However,all the conventional antitumor agents showed no
inhibitory effect. Conclusion: 1The screening method of DNA primase inhibitors was
established.2 Chloramphenicol and sphingosine have inhibitory effect on primase activity.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
1999年第3期292-294,共3页
Chinese Journal of Cancer
基金
国家自然科学基金
