Musashi1通过调控Wnt/β-catenin影响卵巢上皮性肿瘤细胞化疗敏感性

Musashi1 affects chemosensitivity of ovarian epithelial tumor cells by regulating Wnt/β-catenin

目的 探究神经干细胞RNA结合蛋白Musashi1在卵巢上皮性肿瘤(EOC)中的表达及通过调控Wnt/β-连环蛋白(β-catenin)对癌细胞化疗敏感性的影响。方法 收集2012年10月至2014年4月经手术切除的50例EOC及癌旁组织样本,qRT-PCR、WB检测EOC、癌旁组织及EOC细胞系Musashi1表达水平。通过四甲基偶氮唑盐(MTT)检测、克隆形成、划痕、Transwell实验、蛋白免疫印迹(WB)检测、悬滴培养实验、体内异种移植实验及免疫荧光检测探究Musashi1表达对OVCAR420、SKOV3细胞、Wnt/β-catenin信号通路的影响。结果 Musashi1在EOC组织与细胞系中呈高表达,其高表达与FIGOⅢ~Ⅳ期、TNMⅡ~Ⅲ期、淋巴结转移阳性、腹水肿瘤细胞阳性相关(P<0.05)。抑制Musashi1表达可降低OVCAR420细胞恶性生物学行为、肿瘤细胞干性特征、化疗敏感性、N-cadherin、Vimentin、β-catenin、Snail、Cyclin D1表达水平、皮下肿瘤质量(P<0.05),提高E-cadherin表达水平(P<0.05);过表达Musashi1可增强SKOV3细胞恶性生物学行为、肿瘤细胞干性特征、化疗敏感性、N-cadherin、Vimentin、β-catenin、Snail、Cyclin D1表达水平、皮下肿瘤质量(P<0.05),降低E-cadherin表达水平(P<0.05)。结论 Musashi1在EOC中高表达,其高表达与不良临床病理特征相关,可能通过调控Wnt/β-catenin信号通路增强肿瘤细胞恶性生物学行为、提升肿瘤细胞干性特征、化疗敏感性等方面促进EOC的发展。

Objective To explore the expression of RNA binding protein Musashi1 of neural stem cells in epithelial ovarian tumors(EOC)and the effect of Wnt/β-catenin(β-catenin)on chemosensitivity of cancer cells.Methods Collected 50 EOC and adjacent tissue samples from surgical resection from October 2012 to April 2014.Detection of Musashi1 expression in EOC,paracancerous tissues and EOC cell lines by qRT-PCR and WB.The effects of Musashi1 expression on OVCAR420,SKOV3 cells and Wnt/β-catenin signal pathway were investigated by methyl thiazolyl tetrazolium(MTT)assay,clone formation,scratch,Transwell test,western blotting(WB),hanging drop cell spheroid culture,in vivo xenotransplantation test and immunofluorescence detection.Results Musashi1 was highly expressed in EOC tissues and cell lines,and its high expression was related to FIGO stageⅢ–Ⅳ,TNM stageⅡ–Ⅲ,lymph node metastasis and ascites tumor cells(P<0.05).Inhibition of Musashi1 expression could decrease OVCAR420 cell malignant biological behavior,tumor cell dryness,chemosensitivity,N-cadherin,Vimentin,β-catenin,Snail,Cyclin D1 expression,subcutaneous tumor quality(P<0.05),and increase E-cadherin expression(P<0.05).Overexpression of Musashi1 could enhance SKOV3 cell malignant biological behavior,tumor cell dryness,chemosensitivity,N-cadherin,Vimentin,β-catenin,Snail,Cyclin D1 expression,subcutaneous tumor quality(P<0.05),and decrease E-cadherin expression(P<0.05).Conclusion Musashi1 was highly expressed in EOC,and its high expression was related to poor clinicopathological features.It may promote the development of EOC by affecting Wnt/β-catenin signal pathway to enhance the malignant biological behavior of tumor cells,enhance tumor cell dryness,and sensitivity to chemotherapy.