PMID: 11931702Apr 5, 2002Paper

Effect of phorbol esters on activity of Ha-ras gene promoter in HeLa cells

Acta Pharmacologica Sinica
Wei ZhangPing Gao

Abstract

To study the role of phorbol esters (PMA) in the activity of promoter of Ha-ras in HeLa cells. After treatment with PMA, the growth rate of HeLa cells was measured by MTT assay and the expression of Ha-ras gene was detected by reversed transcriptase polymerase chain reaction. The regulation fragment of Ha-ras was constructed into the plasmid enhanced yellow fluorescent protein (EYFP) which did not contain promoter element. The recombinant plasmid pRasEYFP was transient transfected to HeLa cells. Using the EYFP gene as reporter, the activity of promoter of Ha-ras after treatment of PMA was assayed. The growth rate of PMA-treated HeLa cells was markedly reduced compared with untreated cells. The expression of Ha-ras gene was obviously decreased and the activity of promoter of Ha-ras was decreased by 34.0 %and 26.7 % respectively in HeLa cells after treated with PMA (100 microg/L) for 48 h and 72 h. In the meanwhile the activity of protein kinase C (PKC) was decreased by 76.3 % and 73.2 % compared with the control cells, respectively. PMA play an important role in regulation of the activity of promoter of Ha-ras in HeLa cells. The molecular mechanism may be through the PKC pathway.

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