摘要
目的探究胰岛素样生长因子2 mRNA结合蛋白2(IGF2BP2)对结直肠癌细胞增殖、迁移能力和肿瘤免疫微环境的影响及其可能的分子机制.方法使用癌症基因图谱(TCGA)数据库分析结直肠癌及癌旁组织的IGF2BP2和MYC表达水平.采用RNA干扰技术(RNAi)沉默HCT-116和SW480人结直肠癌细胞的IGF2BP2表达,实时定量PCR检测沉默效果.敲低IGF2BP2后,集落形成实验、CCK-8实验、5-乙炔基-2'-脱氧尿苷(EdU)实验检测细胞集落形成和增殖能力,Transwell^(TM)实验检测细胞迁移能力,实时定量PCR检测IGF2BP2、MYC、肿瘤坏死因子α(TNF-α)、转化生长因子β(TGF-β)、白细胞介素10(IL-10)的mRNA表达.Western blot法测定IGF2BP2、MYC的蛋白表达,RNA免疫共沉淀后实时定量PCR(RIP-qPCR)检测HCT-116细胞中IGF2BP2与MYC mRNA结合能力.结果TCGA数据库结果表明,IGF2BP2和MYC在结直肠癌组织中的表达显著高于癌旁组织,且IGF2BP2高表达的结直肠癌患者生存期更短.沉默IGF2BP2后,HCT-116和SW480细胞的活力、增殖、迁移能力降低.IGF2BP2敲低组MYC、TGF-β、IL-10的mRNA表达显著降低,而TNF-αmRNA表达升高;MYC蛋白表达及mRNA稳定性均显著下降.RIP-qPCR结果表明IGF2BP2能够与MYC mRNA结合.结论干扰IGF2BP2下调MYC表达抑制结直肠癌细胞增殖、迁移并促进肿瘤免疫.
Objective To investigate the effect of insulin-like growth factor 2 mRNA binding protein 2(IGF2BP2)on the proliferation,migration and tumor immune microenvironment of colorectal cancer cells and its possible molecular mechanism.Methods The Cancer Genome Atlas(TCGA)database was used to analyze the expression levels of IGF2BP2 and MYC in colorectal cancer and adjacent tissues.The expression of IGF2BP2 in HCT-116 and SW480 human colorectal cancer cells was silenced by RNA interference(RNAi),and the silencing effect was detected by quantitative real-time PCR.After knocking down IGF2BP2,colony formation assay,CCK-8 assay and 5-ethynyl-2'-deoxyuridine(EdU)assay were employed to detect cell colony formation and proliferation ability.TranswellTM assay was used to detect cell migration ability.Quantitative real-time PCR was used to detect the mRNA expression of IGF2BP2,MYC,tumor necrosis factor-α(TNF-α),transforming growth factor-β(TGF-β)and interleukin-10(IL-10).The protein expression of IGF2BP2 and MYC was detected by western blot.The binding ability of IGF2BP2 and MYC in HCT-116 cells was detected by quantitative real-time PCR after RNA immunoprecipitation.Results The results of TCGA database showed that the expression of IGF2BP2 and MYC in colorectal cancer tissues was significantly higher than that in adjacent tissues,and the survival time of colorectal cancer patients with high expression of IGF2BP2 was shorter.After silencing IGF2BP2,the viability,proliferation and migration of HCT-116 and SW480 cells were decreased.The mRNA expression of MYC,TGF-βand IL-10 in IGF2BP2 knockdown group was significantly decreased,while the expression of TNF-αmRNA was increased.The expression of MYC protein and the stability of MYC mRNA were significantly decreased.RIP-qPCR results showed that IGF2BP2 could bind to MYC mRNA.Conclusion Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation,migration and promotes tumor immunity by down-regulating MYC expression.
作者
刘天玥
韩晨颖
胡尘辰
毛司懿
孙元杰
杨舒雅
杨琨
LIU Tianyue;HAN Chenying;HU Chenchen;MAO Siyi;SUN Yuanje;YANG Shuya;YANG Kun(Department of Immunology,The Fourth Regiment,Basic Medical Science Academy,Air Force Medical University,Xi'an 710032,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2023年第4期303-310,共8页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(82073154,82103461)
陕西省重点研发计划项目(2022SF-042)。
