靶向融合肽对裸鼠顺铂耐药卵巢癌移植瘤生长的抑制作用及其机制

Inhibitory effect of MAP2K6-FP on growth of cisplatin-resistant ovarian cancer xenografts in nude mice

目的 观察靶向融合肽(MAP2K6-FP)对裸鼠顺铂耐药卵巢癌移植瘤生长的影响,并探讨其可能机制。方法 取裸鼠14只,每只裸鼠腹腔内注射200μL的SKOV3/DDP细胞(5×106/mL)制备顺铂耐药卵巢癌移植瘤模型,注射1周后,将制模成功的裸鼠随机分为两组(各7只)。观察1组尾静脉注射0.25 mg/kg的MAP2K6-FP,每周2次;对照1组尾静脉注射5 mL/kg的生理盐水,每周2次;两组均注射4周。腹腔注射细胞5周后,统一处死全部裸鼠。比较两组裸鼠腹围、腹水量、转移器官数、瘤结节数、瘤体重量及移植瘤中血管内皮生长因子特异性受体3(VEGFR-3)、淋巴管内皮透明质酸受体-1(LYVE-1)、唾液酸糖蛋白(D2-40)、表皮生长因子受体3(EGFR3)。另取受精后4 d的斑马鱼卵,随机分成两组各20尾,观察2组加入20 mmol/L的MAP2K6-FP,对照2组加入等量生理盐水,1 h后比较两组斑马鱼淋巴管生成情况。结果 与对照1组比较,观察1组腹围、腹水量、转移器官数、瘤结节数、瘤体重量减少及移植瘤中VEGFR-3、LYVE-1、D2-40、EGFR3表达降低(P均<0.05)。观察2组鱼腹部不能形成连续的管腔,对照2组鱼腹部可见三条管腔伴行。结论 MAP2K6-FP可以抑制裸鼠顺铂耐药卵巢癌移植瘤生长,机制可能是抑制淋巴管生成;斑马鱼淋巴管生成受MAP2K6-FP抑制可进一步印证上述机制。

Objective To investigate the inhibitory effect of Mitogen-activated protein kinase kinase 6-fusion protein(MAP2 K6-FP) on the growth of cisplatin-resistant ovarian cancer xenografts in nude mice and to explore its possible mechanism.Methods Fourteen female nude mice were injected with 200 μL SKOV3/DDP cells(5×10~6/mL) to establish cisplatin-resistant xenografted ovarian tumor.At 1 week after injection,the nude mice were divided into two groups:the observation group 1 and control group 1,with 7 nude mice in each group.The observation group 1 was injected with0.25 mg/kg MAP2 K6-FP,twice a week through tail vein injection,and the control group 1 was injected with 5 mL/kg normal saline,twice a week,and both groups were injected for 4 weeks.The nude mice were sacrificed at 5 weeks after injection.The abdominal circumference,ascites volume,number of metastatic organs,number of tumor nodules,tumor body weight,and vascular endothelial growth factor-specific receptor 3(VEGFR-3),lymphatic vessel endothelial hyaluronan receptor 1(LYVE-1),sialoglycoprotein(D2-40),and epidermal growth factor receptor 3(EGFR3) were compared between these two groups.Forty zebra fish at 4 d after fertilization were divided into two groups with twenty zebra fish in each group.The observation group 2 was treated with 20 mmol/L MAP2 K6-FP,and the control group 2 was treated with the same amount of normal saline.After 1 h,the lymphangiogenesis was observed in the two groups.Results Compared with the control group 1,the abdominal circumference,ascites volume,number of metastatic organs,number of tumor nodules,and tumor body weight decreased,and the expression levels of VEGFR-3,LYVE-1,D2-40,and EGFR3 decreased in the observation group 1(all P<0.05).The continuous lumen could not form in the observation group 2,but three lumens were seen in the abdomen of zebra fish in the control group 2.Conclusions MAP2 K6-FP can inhibit the growth of cisplatin-resistant ovarian cancer xenografts in nude mice,possibly by inhibiting lymphangiogenesis.The inhibition of lymphangiogenesis by MAP2 K6-FP in zebra fish can further confirm the above mechanism.