摘要
目的探讨腹膜间皮细胞(HPMC)对卵巢癌细胞血管生成因子表达及分泌的影响。方法用ELISA法检测HPMC条件培养液中肿瘤坏死因子α(TNF-α)及白介素-1β(IL-1β)的水平。用培养小室Millicell将卵巢癌细胞SKOV3与HPMC在不同培养条件下进行共培养。用RT-PCR方法检测SKOV3血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的基因表达,ELISA法检测SKOV3条件培养液中VEGF和bFGF的蛋白水平。结果HPMC条件培养液中可检测到TNF-α和IL-1β。SKOV3与HPMC共培养后,其VEGF和bFGF mRNA表达增强,条件培养液中VEGF和bFGF蛋白水平升高,与SKOV3单独培养相比,差异均有统计学意义(P<0.01)。加入TNF-α或IL-1β的中和抗体共培养,可明显抑制SKOV3 VEGF及bFGF mRNA表达及蛋白分泌(P<0.01),共培养体系中同时加入TNF-α中和抗体及IL-1β中和抗体时,抑制作用增强(P<0.05)。结论HPMC分泌TNF-α和IL-1β,刺激卵巢癌细胞表达及分泌更高的VEGF和bFGF,参与卵巢癌的血管生成及腹膜转移。
Objective To investigate the impact of human peritoneal mesothelial cells (HPMC) on angiogenesis factor expression and secretion of ovarian carcinoma cell line SKOV3. Methods The conditioned medium with HPMC was tested by ELISA for tumor necrosis factor-α (TNF-α)and interleukin 1 β (IL-Iβ). Millicell was used to co-culture HPMC and ovarian carcinoma cell line SKOV3 in the presence or absence of neutralizing antibody against TNF-α or IL-1β. RT-PCR was used to detect vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) gene expression in SKOV3 cells. VEGF and bFGF protein levels in the SKOV3 conditioned medium were assessed by ELISA. Results Conditioned medium with HPMC contained both TNF-α and IL-1β. SKOV3 co-cultured with HPMC expressed higher levels of VEGF and bFGF mRNA and secreted at increased levels of both VEGF and bFGF, in comparison with those in SKOV3 cells cultured alone( P 〈 0.01 ). Addition of neutralizing antibody against TNF-α or IL- 1 β during co-cultures resulted in decrease in mRNA expression and secretion of VEGF and bFGF in SKOV3 cells. When both antibodies were administered during co-cuhure, additive decrease was observed. Conclusion HPMC can act in a paracrine fashion to stimulate ovarian tumor cells to produce and secret at increased levels of VEGF and bFGF through TNF-α and IL-1β, and contribute to angiogenesis and peritoneal metastasis of ovarian cancer.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2006年第10期737-740,共4页
Chinese Journal of Oncology
关键词
卵巢肿瘤
腹膜间皮细胞
血管生成因子
腹膜
转移
Ovarian neoplasms
Peritoneal mesothelial cells
Angiogenesis factor
Peritoneum
Metastasis