中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2010年
1期
40-43
,共4页
张慧敏%曹莹%王卫国%李波兰%沈会芳%林壁润%庞建新%吴曙光
張慧敏%曹瑩%王衛國%李波蘭%瀋會芳%林壁潤%龐建新%吳曙光
장혜민%조형%왕위국%리파란%침회방%림벽윤%방건신%오서광
放线菌发酵液%高通量筛选%端粒酶抑制剂%酵母三杂交系统%96孔板%抗肿瘤活性
放線菌髮酵液%高通量篩選%耑粒酶抑製劑%酵母三雜交繫統%96孔闆%抗腫瘤活性
방선균발효액%고통량사선%단립매억제제%효모삼잡교계통%96공판%항종류활성
actinomycetes fermentation broth%high-throughput screening%telomerase inhibitors%yeast three-hybrid system%96-well plates%anti-tumor activity
目的 从2 000多株放线菌发酵液中高通量筛选特异性端粒酶抑制剂.方法 经对持有的3种端粒酶抑制剂筛选模型即3株酵母菌PCR验证后,将其接种于含3-AT、组氨酸缺乏的酵母培养液,然后分别调整其菌液浓度和筛选样品剂量,以备高通量筛选.将酵母菌液移于96孔板,加入待测样品,振摇培养并定时经多功能微孔板分析仪进行定量分析,选取抑菌率大于0.45的样品进行复筛.结果 经验证所持有的3种端粒酶抑制剂筛选模型均可用于下一步的筛选,并选取酵母菌液OD_(595nm) 0.04做为起始筛选浓度,选取放线菌发酵液剂量为5 ml,通过对放线菌发酵液进行初筛和复筛,发现了14株放线菌发酵液的抑菌效率超过0.45.结论 该研究首次高通量筛选端粒酶抑制剂,方法简便快捷,筛选结果稳定可靠,并筛选到14株放线菌发酵液可能具有抗肿瘤活性,为进一步研究和开发抗肿瘤新药奠定基础.
目的 從2 000多株放線菌髮酵液中高通量篩選特異性耑粒酶抑製劑.方法 經對持有的3種耑粒酶抑製劑篩選模型即3株酵母菌PCR驗證後,將其接種于含3-AT、組氨痠缺乏的酵母培養液,然後分彆調整其菌液濃度和篩選樣品劑量,以備高通量篩選.將酵母菌液移于96孔闆,加入待測樣品,振搖培養併定時經多功能微孔闆分析儀進行定量分析,選取抑菌率大于0.45的樣品進行複篩.結果 經驗證所持有的3種耑粒酶抑製劑篩選模型均可用于下一步的篩選,併選取酵母菌液OD_(595nm) 0.04做為起始篩選濃度,選取放線菌髮酵液劑量為5 ml,通過對放線菌髮酵液進行初篩和複篩,髮現瞭14株放線菌髮酵液的抑菌效率超過0.45.結論 該研究首次高通量篩選耑粒酶抑製劑,方法簡便快捷,篩選結果穩定可靠,併篩選到14株放線菌髮酵液可能具有抗腫瘤活性,為進一步研究和開髮抗腫瘤新藥奠定基礎.
목적 종2 000다주방선균발효액중고통량사선특이성단립매억제제.방법 경대지유적3충단립매억제제사선모형즉3주효모균PCR험증후,장기접충우함3-AT、조안산결핍적효모배양액,연후분별조정기균액농도화사선양품제량,이비고통량사선.장효모균액이우96공판,가입대측양품,진요배양병정시경다공능미공판분석의진행정량분석,선취억균솔대우0.45적양품진행복사.결과 경험증소지유적3충단립매억제제사선모형균가용우하일보적사선,병선취효모균액OD_(595nm) 0.04주위기시사선농도,선취방선균발효액제량위5 ml,통과대방선균발효액진행초사화복사,발현료14주방선균발효액적억균효솔초과0.45.결론 해연구수차고통량사선단립매억제제,방법간편쾌첩,사선결과은정가고,병사선도14주방선균발효액가능구유항종류활성,위진일보연구화개발항종류신약전정기출.
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.
