卵巢癌细胞诱导腹膜间皮细胞血管内皮生长因子表达增强

Up-expression of vascular endothelial growth factor in human peritoneal mesothelial cells induced by ovarian carcinoma cells

目的探讨卵巢癌细胞对腹膜间皮细胞（HPMC）血管内皮生长因子（VEGF）表达的影响.方法以卵巢癌细胞SKOV3条件培养液（SKOV3-CM）培育HPMC 3～72 h,在不同时间点以RT-PCR检测HPMC VEGF基因表达,用ELISA检测HPMC裂解液中VEGF蛋白水平.以SKOV3-CM或SKOV3-CM加转化生长因子β1（TGF-β1）中和抗体培育HPMC 24 h,检测其VEGF基因及蛋白表达的变化.结果在SKOV3-CM未培育的HPMC中检测到VEGFmRNA及蛋白的表达.HPMC在SKOV3-CM中培育6及12 h时,VEGF mRNA及蛋白表达开始增加（P＜0.01）,并以时间依赖性表达增强（P＜0.01）,分别在24及48 h达高峰,48及72 h达到平台期.TGF-β1的中和抗体可部分阻断SKOV3-CM对HPMC VEGF mRNA及蛋白表达的诱导作用（P＜0.01）.结论HPMC可合成VEGF,有利于卵巢癌的腹膜转移和腹水形成.卵巢癌细胞分泌的TGF-β1诱导HPMC VEGF表达升高,间接的促进自身腹膜播散.

Objective To investigate the impact of ovarian carcinoma cells on the expression of vascular endothelial growth factor（VEGF） in human peritoneal mesothelial cells（HPMC）. Methods HPMC was incubated with ovarian carcinoma cell line in SKOV3 conditioned medium（SKOV3-CM） for（3 -72 hrs）. RT-PCR was used to detect VEGF gene expression. VEGF protein expression in cell lysates was assessed by ELISA. HPMC was cultured for 24 hrs in SKOV3-CM or SKOV3-CM with anti-transforming growth factorβ1 （ TGF-β1 ） neutralizing antibody, and the changes in VEGF gene and protein expression were examined. Results VEGF mRNA and protein were detec-ted in HPMC without stimulation of SKOV3-CM. SKOV3-CM significantly induced HPMC VEGF mRNA and protein expression in a time-dependent manner and the induction started 6h（ for mRNA） and 12 h（ for protein） after the treatment（P 〈 0. 01 ）. VEGF mRNA and protein expression achieved maximum level around 24 h and 48 h, and then moved to a plateau at 48 h and 72 h respectively. TGF-β1 neutralizing antibody attenuated SKOV3-CM stimulated VEGF mRNA and protein expression in HPMC. Conclusions HPMC contributes to the development of metastases and the accumulation of malignant ascites of ovarian carcinoma due to the production of VEGF. Ovarian carcinoma cells can regulate VEGF expression in HPMC through TGF-β1, and to enhance peritoneal dissemination indirectly.