ABSTRACT
BACKGROUND: Cervical cancer is one of the most common causes of cancer-associated mortality among affected women in the world. At present, treatment with weekly cisplatin plus ionizing radiation (IR) therapy is the standard regimen for cervical cancer, especially for locally advanced cervical cancer. The purpose of this study is to determine whether FEN1 inhibitors could enhance the therapeutic effect of IR therapy. METHODS: Western blot was applied to determine the expression of FEN1- and apoptosis-related proteins. Cell growth inhibition assay and colony formation assay were used to determine the effects of FEN1 inhibitor and IR exposure for Hela cells in vitro. CRISPR technology was used to knockdown FEN1 expression level of 293T cells, and tumor xenograft in nude mice was employed to determine the effects of FEN1 inhibitor and IR exposure on tumor growth in vivo. RESULTS: Our data revealed that FEN1 is overexpressed in HeLa cell and can be upregulated further by IR. We also demonstrated that FEN1 inhibitor enhances IR sensitivity of cervical cancer in vitro and in vivo. CONCLUSION: FEN1 inhibitor SC13 could sensitize radiotherapy of cervical cancer cell.
Subject(s)
Enzyme Inhibitors/pharmacology , Flap Endonucleases/antagonists & inhibitors , Radiation Tolerance/drug effects , Radiation-Sensitizing Agents/pharmacology , Animals , Apoptosis/drug effects , Apoptosis/radiation effects , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/radiation effects , Disease Models, Animal , Female , Flap Endonucleases/genetics , Flap Endonucleases/metabolism , Gene Expression , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/radiation effects , HeLa Cells , Humans , Mice , Radiation, Ionizing , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/metabolism , Uterine Cervical Neoplasms/pathology , Xenograft Model Antitumor AssaysABSTRACT
The aim of the present study was to explore an optimal method for maturation induction of dendritic cells (DCs). Human monocyte-derived DCs were induced in the presence of GM-CSF and IL-4. On Day 6, the maturation of DCs was induced with CD40L, LPS, TNF-α and cocktail of cytokines (TNF-α, IL-6, IL-1ß and PGE2), respectively, for 24 h. Then, DCs were harvested and subjected to flow cytometry (FCM) for the detection of CD80, CD83, CD86 and HLA-DR. FITC-dextran endocytic activity was measured by FCM, IL-12 production by ELISA and T lymphocyte proliferation following DC stimulation by MTT assay. CD40L, LPS, TNF-α and a cocktail of cytokines induced DC maturation. Induction with the cocktail of cytokines was the most efficient, and the expression rate of CD83 was 66.91% (P<0.05). The FITC-dextran endocytic activity of mature DCs was significantly reduced, and IL-12 production was dramatically increased in mature DCs, particularly in those following induction using the cocktail of cytokines. The mature DCs had potent ability to stimulate the proliferation of lymphocytes. The cocktail of cytokines is a favorable strategy for the induction of DC maturation.
