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Downregulation of p16ink4a inhibits cell proliferation and induces G1 cell cycle arrest in cervical cancer cells

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Studies have suggested that p16ink4a may be a surrogate biomarker for the diagnosis of cervical cancer; however, the function of p16ink4a in human cervical cancer cells remains largely unknown. Therefore, in this study, we aimed to investi­gate the role of p16ink4a in human cervical cancer cells. Immunocytochemistry was used to examine invasive squamous cell carcinoma and its precancerous lesions. p16ink4a-siRNA was transfected into SiHa and HeLa cells to deplete its expression. The cellular levels of p16ink4a mRNA and protein were detected by qRT-PCR and western blot analysis. Proliferation rates were assessed by methyl thiazolyl tetrazolium (MTT) and plate colony formation assays. Cellular migration and invasion ability were assessed by a wound healing assay and Transwell assay. Cellular apoptosis and the cell cycle were measured by flow cytometry. The protein levels of retinoblastoma (Rb), phosphorylated Rb (phospho-Rb), cyclin D1 and caspase-3 were determined by western blot analysis. The results revealed that p16ink4a was overexpressed in the cervical cancer and precancerous lesions (P<0.05). The downregulation of p16ink4a in the SiHa and HeLa cells inhibited their proliferation, migration and invasion. In the SiHa cells, p16ink4a-siRNA also induced G1 cell cycle arrest and apoptosis. Western blot analysis revealed that the downregulation of p16ink4a in the SiHa cells markedly induced caspase-3 activation and decreased cyclin D1 expression. These data suggest that the overexpression of p16ink4a appears to be useful in monitoring cervical precancerous lesions, which supports that the hypothesis that p16ink4a is a surrogate biomarker for the diagnosis of cervical cancer. The therapeutic targeting of overexpressed p16ink4a in the p16ink4a-cyclin-Rb pathway may be a useful strategy in the treatment of cervical cancer.
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Document Type: Research Article

Affiliations: Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital of China Welfare Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, P.R. China

Publication date: June 1, 2014

More about this publication?
  • The International Journal of Molecular Medicine is a monthly, peer-reviewed journal devoted to the publication of high quality studies related to the molecular mechanisms of human disease. The journal welcomes research on all aspects of molecular and clinical research, ranging from biochemistry to immunology, pathology, genetics, human genomics, microbiology, molecular pathogenesis, molecular cardiology, molecular surgery and molecular psychology.

    The International Journal of Molecular Medicine aims to provide an insight for researchers within the community in regard to developing molecular tools and identifying molecular targets for the diagnosis and treatment of a diverse number of human diseases.
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