Upregulated insulin receptor tyrosine kinase substrate promotes the proliferation of colorectal cancer cells via the bFGF/AKT signaling pathway  

胰岛素受体酪氨酸激酶底物过表达可通过bFGF/AKT信号通路促进结直肠癌细胞增殖

作　　者：Song Wang Zheng Liu Yi-Ming Ma Xu Guan Zheng Jiang Peng Sun En-Rui Liu Yu-Kun Zhang Hong-Ying Wang Xi-Shan Wang 

机构地区：[1]Department of Colorectal Surgery,The Second Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang,P.R.China [2]Department of Colorectal Surgery,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing,P.R.China [3]State Key Laboratory of Molecular Oncology,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing,P.R.China

出　　处：《Gastroenterology Report》2021年第2期166-175,I0002,I0003,共12页胃肠病学报道（英文）

基　　金：supported by the National Program Project for Precision Medicine in National Research and Development Plan of China [No.2016YFC0905300];National Natural Science Foundation of China [No.81572930];National Key Research and Development Program of the Ministry of Science and Technology of China [No.2016YFC0905303];Beijing Science and Technology Program [No.D171100002617004].

摘　　要：Background:Some recent studies on insulin receptor tyrosine kinase substrate(IRTKS)have focused more on its functions in diseases.However,there is a lack of research on the role of IRTKS in carcinomas and its mechanism remains ambiguous.In this study,we aimed to clarify the role and mechanism of IRTKS in the carcinogenesis of colorectal cancer(CRC).Methods:We analysed the expression of IRTKS in CRC tissues and normal tissues by researching public databases.Cancer tissues and adjacent tissues of 67 CRC patients who had undergone radical resection were collected from our center.Quantitative real-time polymerase chain reaction and immunohistochemistry were performed in 52 and 15 pairs of samples,respectively.In vitro and in vivo experiments were conducted to observe the effect of IRTKS on CRC cells.Gene Set Enrichment Analysis and Metascape platforms were used for functional annotation and enrichment analysis.We detected the protein kinase B(AKT)phosphorylation and cell viability of SW480 transfected with small interfering RNAs(siRNAs)with or without basic fibroblast growth factor(bFGF)through immunoblotting and proliferation assays.Results:The expression of IRTKS in CRC tissues was higher than that in adjacent tissues and normal tissues(all P<0.05).Disease-free survival of patients with high expression was shorter.Overexpression of IRTKS significantly increased the proliferation rate of CRC cells in vitro and the number of tumor xenografts in vivo.The phosphorylation level of AKT in CRC cells transfected with pLVX-IRTKS was higher than that in the control group.Furthermore,siRNA-IRTKS significantly decreased the proliferation rate of tumor cells and the phosphorylation level of AKT induced by bFGF.Conclusion:IRTKS mediated the bFGF-induced cell proliferation through the phosphorylation of AKT in CRC cells,which may contribute to tumorigenicity in vivo.背景:最近关于胰岛素受体酪氨酸激酶底物(IRTKS)的研究主要聚焦于其在疾病中的功能。然而,鲜有研究探讨IRTKS在肿瘤发生中的作用,其机制亦不明确。本研究旨在阐明IRTKS在结直肠癌(CRC)发生中的作用及机制。方法:通过公共数据库比较IRTKS在CRC组织和正常组织的表达,并收集本中心67例行CRC根治性切除手术患者的癌组织和癌旁组织。利用实时定量聚合酶链式反应(qRT-PCR)和免疫组化分别检测52对冷冻样本的IRTKS mRNA水平以及15对石蜡包埋样本的蛋白质表达。通过体外和体内研究观察IRTKS对CRC细胞的作用。利用GSEA和Metascape平台对测序结果进行功能注释和富集分析。将带有或不带有碱性成纤维细胞生长因子(bFGF)的siRNA转染SW480细胞,通过免疫印迹和增殖实验检测转染后细胞的蛋白激酶B(AKT)磷酸化情况及细胞活性。结果:CRC组织的IRTKS表达量显著高于癌旁组织和正常肠黏膜组织(均P<0.05),且IRTKS高表达患者无病生存率相对较低。IRTKS过表达不仅能够促进CRC细胞的体外增殖,而且能够增加体内移植瘤的数量。转染pLVX-IRTKS质粒的CRC细胞AKT磷酸化水平高于对照组。而且,siRNA-IRTKS可抑制bFGF诱导的细胞增殖和AKT磷酸化。结论:IRTKS通过调节AKT磷酸化促进bFGF诱导的CRC细胞增殖,这一机制可能导致体内成瘤。

关 键 词：colorectal cancer PROLIFERATION IRTKS AKT BFGF 

分 类 号：R735.37[医药卫生—肿瘤]