Transforming growth factor-β1 induces apoptosis through down-regulation of c-myc gene and overexpression of p27Kip1 protein in cervical carcinoma

Transforming growth factor-beta 1 (TGF-beta 1) is known to be a potent growth inhibitor for many cell types, including most epithelial cells. In skin keratinocytes, TGF-beta 1 has been shown to inhibit growth and to rapidly reduce c-myc expression. However, the molecular mechanism of TGF-beta 1 action on cell growth of cervical carcinoma has not yet been elucidated. We thus assessed the effect of TGF-beta 1 on the growth of cervical carcinoma cell lines. Two cervical squamous carcinoma cell lines, CUMC-3 and CUMC-6, were incubated with varying concentrations of TGF-beta 1, and growth inhibition was evaluated with tetrazolium-based colorimetric assay. After culture in TGF-beta 1 for 24 h, inhibition of growth was detected in a dose-dependent manner at concentrations of 0.1-10 ng/ml in both cell lines. This effect of TGF-beta 1 on cultured carcinoma cells was associated with apoptotic process including oligonucleosomal ladder DNA and apoptotic body formations. Northern blot analysis revealed c-myc mRNA expression was suppressed by 10 ng/ml of TGF-beta 1 following 3 h of treatment in both cell lines. Western blot analysis showed that the level of p27(Kip1) protein was increased after TGF-beta 1 treatment in both cell lines. These results suggest that the mechanisms by which TGF-beta 1 inhibits the growth of cervical carcinoma are complex and may include effects on down-regulation of c-myc gene, and overexpression of p27(Kip1) protein. (C) 1998 Academic Press.