[Establishment of high-throughput drug screening cell models based on JAK-STAT signal pathway].
OBJECTIVE
To discover new drugs which may be applied to diseases of the immune system, hemogenesis system diseases and tumors, several high-throughput drug screening cell models based on JAK-STAT signal pathway have been established.


METHODS
Four repeats of STAT DNA binding conserved sequences were synthesized, subcloned into pGL-Luc reporter vector and stably transfected into cell lines in vitro. Cell clones with high copy numbers of STAT binding sites and reporter genes were chosen as high-throughput drug screening cell models. The cell models were tested with known anti-allergic drugs and anti-tumor drugs by determining luciferase activity. The reaction was performed in 96 well micro-plates with a final volume of 50 microL.


RESULTS
The cell models by performing rapid fluorescence assay were shown to be highly sensitive and stable after testing with cytokine and drugs. The modification of the expression plasmid simplified this method and made it more practical. It also provided good linear correlation, wide range of assay, highly sensitive and good reproducibility.


CONCLUSIONS
The method can be performed by high-throughput drug screening for effective extraction of Chinese traditional herbs.
Huang
Mei
M
Institute of Biomedicine, Tsinghua Univiersity, Beijing 100084 , China.
Rong
Yu
Y
Ning
Hong-xiu
HX
Wang
Chun-hua
CH
Wang
Yin-yin
YY
Chang
Zhi-jie
ZJ
D018926:Anti-Allergic Agents / Q000302:isolation & purification / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D053614:Janus Kinase 2; D019169:Jurkat Cells / Q000378:metabolism; D008113:Liver Neoplasms / Q000473:pathology; D008156:Luciferases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D050796:STAT3 Transcription Factor; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D018926:Anti-Allergic Agents; D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; D004365:Drugs, Chinese Herbal; D011518:Proto-Oncogene Proteins; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D015534:Trans-Activators; D008156:Luciferases; D011505:Protein-Tyrosine Kinases; C507924:JAK2 protein, human; D053614:Janus Kinase 2