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An Orthotopic Model of Serous Ovarian Cancer in Immunocompetent Mice for in vivo Tumor Imaging and Monitoring of Tumor Immune Responses
Published on: November 29, 2010
[Study on transfer of interleukin-7 gene into ovarian carcinoma cells in vitro]
Bei Cheng1, Huai-zeng Chen, Da-feng Ye
1Department of Gynecological Tumor, Hospital of Obstetrics and Gynecology, School of Medicine, Zhejiang University, Hangzhou 310006, China.
Objective:
To investigate proliferative activity, expression of cell surface antigens and cytokines of ovarian carcinoma cells transferred by interleukin-7 (IL-7) gene, and its cytotoxic sensitivity to lymphokine activated killer (LAK) cells in vitro through gene transfection technique.
Methods:
Ovarian carcinoma cell line SKOV3 were cultured and transferred by IL-7 cDNA. The proliferation activity of IL-7 gene transferred and nontransferred SKOV3 was observed with inverse phase-contrast light microscope, thiazoyl blue tetrazolium bromide assay and flow cytometry. The expressions of cell surface antigen human leucocyte antigen-ABC (HLA-ABC), human leucocyte antigen-DR (HLA-DR) and intercellular adhesion molecule-I (ICAM-I) were detected by indirect targeted FITC- flow cytometry. The secretions of IL-2, tumor necrosis factor alpha (TNF-alpha) and transforming growth factor (TGF-beta(1)) were assayed by enzyme linked immunosorbent assay. The cytotoxic sensitivity against LAK was detected by LDH release method.
Results:
The morphology, proliferation, cell cycle distributions and HLA-ABC, HLA-DR expression of IL-7 gene transferred SKOV3 didn't change after IL-7 gene transfer. The expression of ICAM-I in IL-7 gene transferred SKOV3 significantly increased and TGF-beta(1) secretion of IL-7 gene transferred SKOV3 significantly decreased. The cytotoxic sensitivity of IL-7 gene transferred SKOV3 to LAK significantly elevated.
Conclusion:
Ovarian carcinoma cell line SKOV3, after transferred by IL-7 gene, remains unchanged proliferation and apoptosis, upregulation of ICAM-I, downregulation of TGF-beta(1) and increases the cytotoxic sensitivity to LAK in vitro.

