摘要
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。
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 inununohistochemical 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. lmnunoprecipitation assay showed that these ligands existed in binding with its receptor. Moneclonal 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 DI/E and p16. Simultaneous adninistration 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 autecrine and an intrac＇rine loop of M - CSF/M - CSFR are present in SMMC 7721 cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2005年第12期2382-2387,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(NO.30270684)
湖南省杰出中青年专家专项基金资助项目(NO.02JJYB004)
湖南省自然科学基金重点项目(NO.04JJ2006)
