cAMP and PMA enhance the effects of IGF-I in the proliferation of endometrial adenocarcinoma cell line HEC-1-A by acting at the G 1 phase of the cell cycle

The present study was undertaken to determine whether endometrial cancer cell line HEC-1-A differ from nontransformed cells, in that the cAMP and protein kinase C pathways may enhance IGF-I effects in mitogenesis by acting at the G 1 phase of the cell cycle instead of G 0 . Immunofluorescence staining of HEC-1-A cells using the proliferating cell nuclear antigen (PCNA) monoclonal antibody and flow cytometric analysis determined that HEC-1-A cells do not enter the G 0 phase of the cell cycle when incubated in a serum-free medium. Approximately 51% of the cells were in G 1 , 12% were in S and 37% in G 2 phase of the cell cycle prior to treatment. Forskolin and phorbol-12-myristate 13-acetate (PMA) were used to stimulate cAMP production and protein kinase C activity, respectively. IGF-I, forskolin and PMA each increased ( P <0.01) [ 3 H]-thymidine incorporation in a dose and time dependent manner. The interaction of forskolin and PMA with IGF-I was then determined. Cells preincubated with forskolin or PMA followed by incubation with IFG-I incorporated significantly more ( P <0.01) [ 3 H]-thymidine into DNA than controls or any treatment alone. It is concluded that forskolin and, to a lesser extent, PMA exert their effect at the G 1 phase of the cycle to enhance IGF-I effects in cell proliferation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75013/1/j.1365-2184.1995.tb00061.x.pd