[Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation, migration and promotes tumor immunity by down-regulating MYC expression].

Tianyue Liu, Chenying Han, Chenchen Hu, Siyi Mao, Yuanjie Sun, Shuya Yang, Kun Yang
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Abstract

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-β and IL-10 in IGF2BP2 knockdown group was significantly decreased, while the expression of TNF-α mRNA 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.

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[敲低IGF2BP2通过下调MYC表达抑制结直肠癌细胞增殖、迁移,促进肿瘤免疫]。
目的探讨胰岛素样生长因子2 mRNA结合蛋白2 (IGF2BP2)对结直肠癌细胞增殖、迁移和肿瘤免疫微环境的影响及其可能的分子机制。方法利用肿瘤基因组图谱(Cancer Genome Atlas, TCGA)数据库分析IGF2BP2和MYC在结直肠癌及癌旁组织中的表达水平。采用RNA干扰(RNAi)方法对HCT-116和SW480人结直肠癌细胞中IGF2BP2的表达进行沉默,并采用实时荧光定量PCR检测其沉默效果。敲除IGF2BP2后,采用集落形成法、CCK-8法和5-乙基-2′-脱氧尿苷(EdU)法检测细胞集落形成和增殖能力。TranswellTM法检测细胞迁移能力。采用实时荧光定量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) mRNA表达。western blot检测IGF2BP2和MYC蛋白的表达。RNA免疫沉淀后,采用实时荧光定量PCR检测IGF2BP2与MYC在HCT-116细胞中的结合能力。结果TCGA数据库结果显示,IGF2BP2和MYC在结直肠癌组织中的表达明显高于癌旁组织,且IGF2BP2高表达的结直肠癌患者生存时间较短。IGF2BP2沉默后,HCT-116和SW480细胞的活力、增殖和迁移能力下降。IGF2BP2敲低组MYC、TGF-β、IL-10 mRNA表达量显著降低,TNF-α mRNA表达量升高。MYC蛋白的表达和MYC mRNA的稳定性均显著降低。RIP-qPCR结果显示IGF2BP2能够结合MYC mRNA。结论IGF2BP2基因下调可抑制结直肠癌细胞的增殖、迁移,并通过下调MYC的表达促进肿瘤免疫。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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