Leptin促进肿瘤相关成纤维细胞活化及卵巢癌腹腔播散

Leptin promotes cancer-associated fibroblast activation and facilitates ovarian cancer peritoneal dissemination

目的探索腹水中瘦素(Leptin)对成纤维细胞活化的作用以及对卵巢癌腹腔播散的影响。方法恶性腹水中分离肿瘤相关成纤维细胞并鉴定;Transwell及系膜撕开试验探究其对卵巢癌细胞侵袭能力的影响;ELISA比较良恶性腹水中Leptin表达水平差异;数据库分析微环境细胞Leptin受体表达情况;免疫荧光、Western blot及胶原回缩试验探究Leptin对成纤维细胞活化的影响;SKOV3原位移植模型探索Leptin对成纤维细胞活化及腹腔播散的影响。结果腹水中肿瘤相关成纤维细胞高表达α-SMA;其显著增加卵巢癌细胞体外侵袭能力;Leptin在良恶性腹水中表达分别为(132.1±15.6)pg/mL、(576.4±60.4)pg/mL;成纤维细胞高表达Leptin受体;Leptin重组蛋白显著增加成纤维细胞活化;移植瘤模型中IgG/Leptin封闭组活化成纤维细胞比例分别为(24.6±3.5)%、(8.5±1.6)%;腹腔种植灶数目分别为(20±3)、(7±2)个;总的转移灶重量分别为(2±0.5)、(0.5±0.1)g。结论卵巢癌腹水中Leptin维持成纤维细胞的活化,可考虑作为遏制卵巢癌腹腔播散的治疗靶点。

Objective To investigate Leptin influence of ascitc fibroblast activation and subsequent ovarian cancer(OC)peritoneal metastasis.Methods Flow cytometry was performed to isolate cancer associated fibroblasts(CAFs)from OC malignant ascites.Transwell and mesothelial clearance assay were performed to explore CAFs influence of OC cells malignant behavior.ELISA was conducted to detect leptin expression in ovarian benigh and malignant ascites.The relative expression of leptin receptor was analyzed in public profile GSE39396.Immunoflurescence,Western blot and collagen contraction assay were conducted to show leptin exertion on fibroblast activation.SKOV3 orthotopic xenograft model was employed to investigate leptin eradication influence of fibroblast activation and metastases formation.Results Ascitic CAFs were characterized by immunostaining ofα-SMA.These cells dramatically enhanced tumor cell invasion and mesothelial clearance.Leptin was notably elevated in malignant ascites(576.4±60.4)pg/mL compared with that of benigh ascites(132.1±15.6)pg/mL.Leptin receptor was distinctly overexpressed in stromal fibroblasts.Human leptin recombinant protein could notably promote fibroblast activation.In SKOV3 xenograft model,PDGFRB+CAFs percentage was(24.6±3.5)%in IgG group compared with(8.5±1.6)%in the leptin neutralizing group.Eventually,the metastatic tumor nodules was(7±2)and tumor burden(0.5±0.1)g in the leptin neutralizing group,while the tumor nodules was(20±3)and tumor burden(2±0.5)g in the IgG group;Conclusions Leptin is involved in the activation of ascitic fibroblasts,and it could be considered as potent therapeutic target in hampering OC peritoneal dissemination