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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 effect of peritoneal mesothelial cells (HPMC) on the expression and secretion of angiogenic factors in ovarian cancer cells. Methods The levels of tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) in HPMC conditioned medium were detected by ELISA. The ovarian cancer cells SKOV3 and HPMC were co-cultured under different culture conditions with a Millicell culture chamber. The gene expressions of VEGF and bFGF in SKOV3 were detected by RT-PCR. The protein levels of VEGF and bFGF in SKOV3 conditioned medium were detected by ELISA. Results TNF-α and IL-1β were detected in HPMC conditioned medium. After co-culture of SKOV3 and HPMC, the expression of VEGF and bFGF mRNA increased, while the level of VEGF and bFGF protein in conditioned medium increased. Compared with SKOV3 alone, the difference was statistically significant (P <0.01). The co-culture of neutralizing antibody with TNF-α or IL-1β significantly inhibited the mRNA and protein secretion of SKOV3 VEGF and bFGF (P <0.01). The co-culture system with simultaneous addition of TNF-α neutralizing antibody and IL- 1β neutralizing antibody, the inhibition was enhanced (P <0.05). Conclusion HPMC secretes TNF-α and IL-1β, stimulates the expression of ovarian cancer cells and secretes higher VEGF and bFGF, which is involved in angiogenesis and peritoneal metastasis of ovarian cancer.