High-throughput screening of telomerase inhibitors in actinomycetes fermentation broth
- VernacularTitle:从放线菌发酵液中高通量筛选端粒酶抑制剂
- Author:
Huimin ZHANG
;
Ying CAO
;
Weiguo WANG
;
Bolan LI
;
Huifang SHEN
;
Birun LIN
;
Jianxin PANG
;
Shuguang WU
- Publication Type:Journal Article
- Keywords:
actinomycetes fermentation broth;
high-throughput screening;
telomerase inhibitors;
yeast three-hybrid system;
96-well plates;
anti-tumor activity
- From:
Chinese Pharmacological Bulletin
2010;26(1):40-43
- CountryChina
- Language:Chinese
-
Abstract:
Aim To discover specific telomerase inhibitors by high-throughput screening in the more than 2 000 strains of actinomycetes fermentation broth.Methods After verifying the three kinds of screening models for telomerase inhibitor by yeast PCR,the three yeast strains were inoculated overnight into yeast liquid culture which was lacking in histidine and contained 3-AT.Then concentration of yeast strains in culture liquid and dosage of screening sample were regulated for better adjustment to the high-throughput screening.At last the adjusted liquid culture containing yeast strain piped to 96-well plates,drug samples and control article were also added,then the yeast was incubated in the thermostat oscillator.At the same time OD_(595nm) of the yeast liquid was measured every 12 h by using Multifunctional Microplate Analyzer,and the samples whose inhibitory efficiency was over 0.45 were selected for secondary screening.Results The high-throughput screening method related to yeast three-hybrid system was effective in screening samples that had potential telomerase inhibitory ability.The initial concentration of yeast liquid OD_(595nm) was 0.04 and the best dosage of samples was 5 ml.14 active samples,whose inhibitory efficiency was over 0.45,were founded after primary and secondary screening.Conclusions High-throughput screening method related to yeast three-hybrid system is simple and stable in discovering telomerase inhibitors,and 14 new antitumor compounds are identified,which lays the foundation in the development of new anti-tumor agents.
