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Transforming Growth Factor-β1 Induces Apoptosis through Down-Regulation of c-mycGene and Overexpression of p27Protein in Cervical Carcinoma
Authors:Jin Woo Kim M.D.  Ho Shik Kim M.D.  In Kyung Kim M.D.  Mee Ran Kim M.D.  Eun Young Cho B.S.  Heung Kee Kim M.D.  Joon Mo Lee M.D.  Sung Eun Namkoong M.D.
Affiliation:aDepartment of Obstetrics and Gynecology, Kangnam St. Mary's Hospital, Seoul, Korea;bMolecular Genetic Laboratory, Catholic Research Institutes of Medical Science, Seoul, Korea;cDepartment of Biochemistry, Catholic University Medical College, 505 Banpo-dong, Seocho-ku, Seoul, 137-040, Korea
Abstract:
Transforming growth factor-β1 (TGF-β1) is known to be a potent growth inhibitor for many cell types, including most epithelial cells. In skin keratinocytes, TGF-β1 has been shown to inhibit growth and to rapidly reduce c-mycexpression. However, the molecular mechanism of TGF-β1 action on cell growth of cervical carcinoma has not yet been elucidated. We thus assessed the effect of TGF-β1 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-β1, and growth inhibition was evaluated with tetrazolium-based colorimetric assay. After culture in TGF-β1 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-β1 on cultured carcinoma cells was associated with apoptotic process including oligonucleosomal ladder DNA and apoptotic body formations. Northern blot analysis revealed c-mycmRNA expression was suppressed by 10 ng/ml of TGF-β1 following 3 h of treatment in both cell lines. Western blot analysis showed that the level of p27Kip1protein was increased after TGF-β1 treatment in both cell lines. These results suggest that the mechanisms by which TGF-β1 inhibits the growth of cervical carcinoma are complex and may include effects on down-regulation of c-mycgene, and overexpression of p27Kip1protein.
Keywords:
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