NFATc1对裸鼠上皮性卵巢癌移植瘤脉管生成的影响

摘要

AIM:To investigate the role of NFATc1 in vascular generation in the nude mice transplanted with human ovarian cancer SKOV3 cells.METHODS: NFATc1 expression was silenced by siRNA in SKOV3 cells.Human ovarian cancer transplantation nude mouse model was established by transplanting with SKOV3 cells in which the NFATc1 gene was silenced by siRNA technique.The expression of NFATc1, CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels was determined by RT-PCR, Western blotting and immunohistochemical staining.The tumor growth, angiogenesis and lymphangiogenesis were also observed.RESULTS:Over-expression of NFATc1 was observed in human ovarian cancer tissues.The silencing of NFATc1 expression by siRNA decreased tumorigenesis of transplanted ovarian cancer cells in the nude mice, reduced tumor vascular generation and inhibited the expression of CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels.CONCLUSION:NFATc1 is overexpressed in ovarian cancer.NFATc1 silencing regulates the tumor vascu-lar generation.NFATc1 thus has potential as a therapeutic target and for use in the diagnosis and evaluating prognosis of epithelial ovarian cancer.%目的：探讨胞浆活化T细胞核因子1（nuclear factor of activated T-cells， cytplasmic 1， NFATc1）对人卵巢癌SKOV3细胞裸鼠移植瘤生长和肿瘤脉管生成的影响及其可能机制。方法：NFATc1 siRNA转染人上皮性卵巢癌细胞株SKOV3，免疫荧光及RT-PCR测量转染效率和基因抑制率，选取效率最高的序列建立裸鼠皮下移植瘤模型，测量各组裸鼠肿瘤体积，观察NFATc1 siRNA的体内抗肿瘤作用。免疫组织化学检测各组肿瘤组织NFATc1的表达情况，并使用细胞角蛋白染色标记上皮性来源，CD34标记微血管，podoplanin标记微淋巴管。分别计算各组微血管及微淋巴管密度并进行统计学分析。应用RT-PCR及Western blot检测各组移植瘤组织NFATc1、CXC趋化因子受体2（CXCR2）、成纤维细胞生长因子2（FGF-2）及血小板源性生长因子BB（PDGF-BB）的mRNA及蛋白表达水平。结果：3条特异性序列均可显著降低NFATc1的表达水平，以siRNA-1169最佳。 NFATc1在空白组及阴性对照组瘤组织高表达。干扰组抑瘤率为57.08％，且重量和体积均低于2个对照组。空白组和阴性对照组的微血管密度和微淋巴管密度明显高于干扰组。对照组比较，NFATc1 siRNA可以在mRNA水平上明显抑制NFATC1、CX-CR2、FGF-2和PDGF-BB的转录。 Western blot各组细胞在相应位置出现NFATc1、CXCR2、FGF-2和 PDGF-BB条带，空白组与阴性对照组的吸光度最强，与干扰组比较具有显著差异。结论： NFATc1 siRNA明显抑制人卵巢癌SKOV3细胞裸鼠皮下移植瘤生长和肿瘤脉管生成，下调CXCR2、FGF-2及PDGF-BB的表达可能为其途径之一。