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KMID : 0613820080180070952
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Journal of Life Science
2008 Volume.18 No. 7 p.952 ~ p.957
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Effect of Various Factors on Early THP-1 Cell Adhesion Induced Phorbol 12-Myristate 13-Acetate (PMA)
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Jo Yong-Sam
Shin Ji-Hyun
Choi Tae-Saeng
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Abstract
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We evaluated the effects of various factors (e.g., serum, inhibitors of protein synthesis, and cytoskeleton and protein kinases) on early PMA-induced THP-1 cell adhesion using an adhesion assay with Sulforhodamine B (SRB) staining, which was used to assess the proliferation of the attached cells. THP-1 cell adhesion to a plastic substrate was detected 1 hr after exposure to Phorbol 12-Myristate 13-Acetate (PMA) and peaked after 18 hr. At concentrations ＞ 25 nM PMA, the level of adhesion did not change. Based on our preliminary results, we used 25 nM PMA and 5 hr of culture as standard assay conditions. Early PMA-induced cell adhesion was not affected by the presence of serum or PD 98059 in the culture medium, but was affected by the addition of PKC inhibitors and cycloheximide. In the presence of actin inhibitor with PMA, the cell adhesion increased when comparing with PMA treatment only. Thus, early PMA-induced adhesion of THP-1 cells does not require serum in the culture medium, MAP-kinase activation, or actin polymerization, but does require de novo protein synthesis and PKC activation. Our SRB-based cell adhesion assay may be used to screen other PKC inhibitors.
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KEYWORD
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Celladhesion, monocyticcells, Phorbol12-myristate13-acetate(PMA), proteinkinaseC
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