色素上皮衍生因子联合顺铂对卵巢癌的抑制作用

Inhibitory effect of pigment epithelium-derived factor combined with cisplatin on ovarian cancer

目的探讨色素上皮衍生因子(PEDF)联合顺铂(DDP)对卵巢癌细胞的抑制作用以及对卵巢癌小鼠的治疗效果。方法体外培养ID8细胞株,将细胞分为对照组、PEDF组和PEDF+DDP组,分别加入PBS、PEDF和PEDF+DDP处理ID8细胞。通过细胞划痕实验和CCK-8实验观察每组细胞迁移和增殖。随后,建立C57BL/6小鼠卵巢癌模型,将24只卵巢癌模型小鼠随机分为模型组、PEDF组和PEDF+DDP组,每组8只。模型组给予100μl PBS,PEDF组给予静脉注射12.5 mg/kg的PEDF蛋白,PEDF+DDP组给予静脉注射12.5 mg/kg PEDF蛋白和腹腔注射2 mg/kg顺铂注射液。隔日给药,共给药3次,20 d后处死小鼠并解剖分离肿瘤,检测肿瘤的体积,通过免疫组化和免疫荧光对肿瘤组织内的新生血管CD31的表达和凋亡细胞进行评价。结果体外细胞实验表明,与对照组相比,PEDF组和PEDF+DDP组ID8细胞的迁移和增殖均显著降低(P<0.05);与PEDF组相比,PEDF+DDP组ID8细胞的迁移和增殖显著降低(P<0.05)。动物体内实验表明,与模型组相比,PEDF组和PEDF+DDP组肿瘤体积的增长速率明显放缓(P<0.05);与PEDF组相比,PEDF+DDP组肿瘤体积的增长速率明显放缓(P<0.05)。与模型组相比,PEDF组和PEDF+DDP组小鼠肿瘤组织中CD31的表达明显下调,而肿瘤细胞的凋亡率明显升高(P<0.05);与PEDF组相比,PEDF+DDP组CD31的表达明显下调,而肿瘤细胞的凋亡率明显提高(P<0.05)。结论PEDF和DDP联合用药对卵巢癌细胞具有协同抑制作用以及对卵巢癌小鼠模型具有协同治疗作用。这种协同治疗效果优于PEDF单独治疗的效果。

Objective To investigate the inhibitory effect of pigment epithelium-derived factor(PEDF)combined with cisplatin(DDP)on ovarian cancer cells and its therapeutic effect on ovarian cancer mouse model.Methods ID8 cells were cultured in vitro,and then divided into three groups:control group,PEDF group and PEDF+DDP group.ID8 cells were treated with PBS,PEDF and PEDF+DDP in three groups,respectively.Cell migration and proliferation in each group were observed by cell scratch test and CCK-8 test.The ovarian cancer model of C57BL/6 mice was established,and 24 ovarian cancer model mice were randomly divided into model group(n=8),PEDF group(n=8)and PEDF+DDP group(n=8).The mice were given 100μl PBS in control group.The mice in PEDF group were intravenously injected with 12.5 mg/kg PEDF protein.The mice in PEDF+DDP group were given intravenous injection of 12.5 mg/kg PEDF protein and intraperitoneal injection of 2 mg/kg cisplatin.The corresponding drugs were given every other day for three times.At 20 d after the last treatment,the mice were executed and the tumor was isolated.The volume of the tumor was measured,and CD31 expression in neovascularization and the apoptotic cells in tumor tissues were evaluated by immunohistochemistry and immunofluorescence.Results The cell experiment in vitro showed that the migration and the proliferation of ID8 cells in PEDF group and PEDF+DDP group were significantly lower than those in control group(P<0.05).Compared with PEDF group,the migration and the proliferation of ID8 cells were significantly decreased in PEDF+DDP group(P<0.05).The experiment in vivo showed that the tumor volume was significantly smaller in PEDF group and PEDF+DDP group than in model group(P<0.05).Compared with PEDF group,the tumor volume in PEDF+DDP group was significantly decreased(P<0.05).Compared with control group,the expression of CD31 in tumor tissue was significantly down-regulated in PEDF group and PEDF+DDP group,while the apoptosis rate of tumor cells was significantly increased(P<0.05).Compared with PEDF group,the CD31 expression was down-regulated and the apoptosis rate of tumor cells was increased in PEDF+DDP group(P<0.05).Conclusion The combination of PEDF and DDP has synergistic inhibitory effect on ovarian cancer cells and synergistic therapeutic effect on ovarian cancer mouse model.The efficacy of synergistic therapy is better than that of PEDF alone.