Proliferation of human hepatoma cells are regulated by the intracrine and autocrine loop of the macrophage colony-stimulating factor system |
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| Authors: | ZHANG Meng-xia LUO Hong-mei TANG Sheng-song LEI Xiao-yong LONG Zhi-feng ZHANG Xiao-hong WANG Yu-hua |
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| Institution: | 1.Medical College,2Division of Pharmacoproteomics, Proteomics Institute of Pharmacy and Pharmacology, Nanhua University, Hengyang 421001, China |
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| Abstract: | AIM: To study the expression and characterization of intracellular macrophage colony-stimulating factor (M-CSF) in human hepatoma cell line, SMMC 7721 cell, and to explore the mechanism by which M-CSF regulates the proliferation of human hepatoma cells. METHODS: The immunohistochemical staining, flow cytometry, antisense technique and Western blotting were used to study the effects and mechanisms of intracellular M-CSF on the proliferation of human hepatoma cells. RESULTS: SMMC 7721 cells highly expressed M-CSF and its receptor. The localization of positive reactions was mainly in cytoplasma and nucleus in SMMC 7721 cells. In cytoplasma and nucleus, one isoforms of M-CSF was found with the molecular weight (MW) of 20 kD, while one type of M-CSFR was discovered with MW of 120 kD. Immunoprecipitation assay showed that these ligands existed in binding with its receptor. Monoclonal antibody (McAb) against M-CSF and antisense oligodeoxynucleotides (ASODN) blocking M-CSF expression inhibited the proliferation of SMMC 7721 cells. McAb and ASODN regulated the expression of cyclin D1/E and p16. Simultaneous administration of both McAb and ASODN inhibited the proliferation of SMMC 7721 cells and modulated the expression of cyclins at greater degrees. CONCLUSION: Our results suggest that an autocrine and an intracrine loop of M-CSF/M-CSFR are present in SMMC 7721 cells. |
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