Notch信号通路在肾母细胞瘤中的表达及其意义

目的 探讨Notch信号通路因子在肾母细胞瘤中的表达及其意义.方法 本研究中实验组为收集2010年6月至2014年3月在我院经手术活检诊断为肾母细胞瘤组织的标本.对照组标本为同期由于肾外伤、重复肾等疾病行手术切除获得.所有病例中,肿瘤组患儿共计31例,男17例,女14例,年龄4～144个月,平均年龄37.4个月,肿瘤分期Ⅰ期9例,Ⅱ期8例,Ⅲ期8例,Ⅳ期6例,尚无Ⅴ期病例.对照组为17例患儿,男10例,女7例,年龄23～157个月,平均年龄95.6个月.首先利用临床收集的肾母细胞瘤及非瘤肾脏组织标本,通过RT-PCR方法检测其中notch1的RNA表达水平;同时通过Western blot方法检测DAPT处理后的SK-NEP-1(人肾母细胞瘤细胞系)中Notch信号通路因子与Wnt信号通路因子,以及VEGF的表达水平并进行统计学分析.结果 不同分期WT中notch1的表达:肿瘤组notch1基因mRNA表达较对照组明显增高,且肿瘤分期越高,notch1基因的mRNA表达也相应越高;DAPT作用SK-NEP-1后的表达:分别利用不同浓度的DAPT作用于SK-NEP-1后,SK-NEP-1中Notch信号通路的notch1、Jagged1蛋白表达水平随DAPT浓度增高逐渐下降,且Wnt信号通路中的wnt1、B-catenin蛋白表达水平同样明显下降,在10μmol/L浓度DAPT时上述蛋白表达量较PBS组比较具有明显统计学差异(P＜0.05);而且利用DAPT抑制之后,SK-NEP-1中的血管内皮生长因子(VEGF)表达也明显降低.结论 Notch信号通路蛋白的高表达与肾母细胞瘤的发生及恶性程度存在关系,同时Notch信号通路作为肿瘤治疗靶点也为我们将来治疗肾母细胞瘤提供新的线索和重要依据.

Expression and significance of Notch signal pathway in Wilm's tumor

Objective To explore the expression and significance of Notch signal pathway factors in Wilm's tumor.Methods The surgical biopsy specimens of Wilrm's tumor were collected from June 2010 to March 2010.And the control group underwent surgical resections due to injuries or renal duplication during the same period.The tumor group had 17 boys and 14 girls with an average age of 37.4 (4-144) months.The stages were Ⅰ (n=9),Ⅱ (n=8),Ⅲ (n=8),Ⅳ (n=6) andV(n=0).And the control group had 10 boys and 7 girls with an average age of 95.6 (23-157) months.After a treatment of N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT),basal expressions of Notch signal pathway factors,Wnt signal pathway factors and VEGF in SK-NEP-1 (human nephroblastoma cell line) were detected by Western blot.Furthermore,mRNA levels of notch1 were detected by real-time polymerase chain reaction (PCR) in Wilm's tumor and non-tumor kidney tissue samples.Results The notch1 mRNA expression was significantly higher in WT group.And the more advanced tumor stage,the higher notch1 mRNA expression.At different concentrations of DAPT,notch1 and Jagged1 protein expression levels in the Notch signaling pathway increased with declining concentrations of DAPT;wnt1,B-catenin in Wnt signaling pathway protein expression levels also dropped markedly.And the protein expression had significant statistical difference in 10 μmol/L of DAPT as compared with PBS group (P＜ 0.05);And after dosing of DAPT inhibitor,the expression of vascular endothelial growth factor (VEGF) also significantly decreased.Conclusions Up-regulated Notch signal pathway factors may be associated with Wilm's tumor and serve as tumor therapeutic targets.