Department of Pediatric Surgery, The First Affiliated Hospital of Zhengzhou University, No. 1 East Jianshe Road, Erqi District, Zhengzhou, 450000, Henan, China.
Department of Respiratory Medicine, The First Affiliated Hospital of Zhengzhou University, No. 1 East Jianshe Road, Erqi District, Zhengzhou, 450000, Henan, China.
J Cancer Res Clin Oncol. 2019 Oct;145(10):2457-2468. doi: 10.1007/s00432-019-03003-0. Epub 2019 Aug 28.
Our previous study identified a Wilms tumor-suppressing peptide (WTSP) that was upregulated in healthy children, but downregulated in children with Wilms tumor (WT). This study aimed to investigate the effect of WTSP on WT growth in vivo and in vitro.
WTSP was synthesized by solid-phase synthesis of FOMC-protected amino acids. Cell growth curve, cytotoxicity, and apoptosis of WTSP-treated human WT cell line (SK-NEP-1) were determined by cell count, Cell Counting Kit-8 assay, and flow cytometry. The expression of key proteins of four WT-associated signaling pathways was determined by real-time PCR and western blotting. The WT xenograft mouse model was established by the armpit injection of SK-NEP-1 cells. The TUNEL assay was used to detect apoptosis in mouse tumor cells.
WTSP inhibited the proliferation of SK-NEP-1 cells in a dose- and time-dependent manner, and it arrested SK-NEP-1 cells in G2/M phase. WTSP-treated cells exhibited a low expression of PCNA and Bcl-2 and high expression of Bax. The expression of β-catenin was markedly changed after WTSP treatment. WTSP-treated mice had significantly smaller tumors than untreated mice.
Our findings indicated an anti-tumor effect of WTSP, which is correlated with Wnt/β-catenin pathway. This newly identified peptide may exert a therapeutic effect of WT in the future.
我们之前的研究发现了一种 Wilms 肿瘤抑制肽（WTSP），它在健康儿童中上调，但在 Wilms 肿瘤（WT）患儿中下调。本研究旨在探讨 WTSP 对 WT 体内和体外生长的影响。
通过 FOMC 保护氨基酸的固相合成合成 WTSP。通过细胞计数、Cell Counting Kit-8 测定和流式细胞术测定 WTSP 处理的人 WT 细胞系（SK-NEP-1）的细胞生长曲线、细胞毒性和细胞凋亡。通过实时 PCR 和 Western blot 测定与四个 WT 相关信号通路相关的关键蛋白的表达。通过 SK-NEP-1 细胞腋窝注射建立 WT 异种移植小鼠模型。TUNEL 检测法用于检测小鼠肿瘤细胞中的凋亡。
WTSP 呈剂量和时间依赖性抑制 SK-NEP-1 细胞的增殖，并使 SK-NEP-1 细胞停滞在 G2/M 期。WTSP 处理的细胞中 PCNA 和 Bcl-2 的表达较低，Bax 的表达较高。WTSP 处理后β-连环蛋白的表达明显改变。WTSP 处理的小鼠肿瘤明显小于未处理的小鼠。
我们的研究结果表明 WTSP 具有抗肿瘤作用，与 Wnt/β-catenin 通路相关。这种新发现的肽可能在未来对 WT 发挥治疗作用。
