肝癌细胞增殖受M-CSF胞内和胞外自分泌的双重调控

目的：研究胞内M-CSF及其受体在肝癌SMMC 7721细胞的表达与性质,探讨胞内M-CSF对SMMC 7721细胞增殖的影响及其机制。 方法： 以高表达M-CSF的人肝癌细胞系（SMMC 7721细胞）为模型，以免疫组化、流式细胞计数、反义技术与蛋白印迹等方法观测胞内M-CSF对SMMC 7721细胞增殖的影响及其机制。 结果： M-CSF 及其受体主要在SMMC 7721细胞的胞质、胞核中表达，胞内的M-CSF的相对分子量为20 000，M-CSFR的相对分子量为120 000；免疫共沉淀分析证明M-CSF在细胞内与M-CSFR以复合物的形式存在；M-CSF的单克隆抗体及其反义寡聚核苷酸能抑制SMMC 7721细胞的增殖、下调cyclinD1/E的表达和上调p16的表达，且M-CSF的单克隆抗体及其反义寡聚核苷酸的联合使用能进一步加强对SMMC 7721细胞抑制作用和增加下调cyclinD1/E和上调p16的表达幅度。 结论： SMMC 7721细胞受M-CSF胞外自分泌和胞内自分泌的双重调控。

Proliferation of human hepatoma cells are regulated by the intracrine and autocrine loop of the macrophage colony-stimulating factor system

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 (A-SODN) blocking M - CSF expression inhibited the proliferation of SMMC 7721 cells. McAb and ASODN regulated the expression of cyclin Dl/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 intiacrine loop of M - CSF/M - CSFR are present in SMMC 7721 cells.