Transforming growth factor-β1 bioassay involving matrix metalloproteinase-2 induction.
The transforming growth factor-β1 (TGF-β1) bioassay developed in this study monitors increased luciferase activity in MCF10A cells containing the matrix metalloproteinase-2 (MMP-2) promoter with a luciferase reporter and treated with increasing TGF-β1 concentrations. The response was linear in the concentration range from 75 to 2,500 pg/mL. The abilities of 3 types of TGF-β in inducing MMP-2 were different. The luciferase activity induced by TGF-β1 was about 2 times more than that by TGF-β2 and TGF-β3. The MMP-2 promoter bioassay showed greater reproducibility (coefficient of variation [CV] 10%) than the previously developed anticell proliferation assay of TF-1 cell (CV 16%) and the MMP-2 zymogram assay (CV 40%).
Kim
Gi-Hyun
GH
Division of Recombinant Product, Biopharmaceutical Bureau, Korea Food and Drug Administration, Eunpyeong-gu Seoul, Republic of Korea.
Kang
So-Young
SY
Kwak
Ho-Jung
HJ
Baek
Kyung-Min
KM
Hong
Seung-Hwa
SH
Lee
Hwa-Jeong
HJ
Sohn
Yeo-Won
YW
D001681:Biological Assay; D002460:Cell Line; D003001:Cloning, Molecular; D004306:Dose-Response Relationship, Immunologic; D004790:Enzyme Induction; D005260:Female; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D049449:Luminescence; D042361:Mammary Glands, Human / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D053773:Transforming Growth Factor beta1; D008156:Luciferases; D020778:Matrix Metalloproteinase 2