慢性饥饿应激通过增强ITGB1表达促进结直肠癌细胞迁移  

Chronic Starvation Stress Promotes Migration Ability of Colorectal Cancer Cells by Inducing ITGB1 Upregulation

作　　者：李思雨 曹静桦 王凤伟 LI Siyu;CAO Jinghua;WANG Fengwei(State Key Laboratory of Oncology in South China,Guangdong Provincial Clinical Research Center for Cancer,Sun Yat-sen University Cancer Center,Guangzhou 510060,China)

机构地区：[1]中山大学肿瘤防治中心,广东省恶性肿瘤临床医学研究中心,华南恶性肿瘤防治全国重点实验室,广州510060

出　　处：《肿瘤防治研究》2024年第4期240-248,共9页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金面上项目(81972227)。

摘　　要：目的探讨慢性饥饿应激对结直肠癌细胞增殖、迁移能力的影响及其机制。方法采用长期血清饥饿法模拟肿瘤细胞慢性饥饿环境,通过梯度降低血清浓度构建结直肠癌SW480、DLD-1低血清耐受亚株并观察细胞形态的改变;利用CCK-8和Transwell实验检测长期血清饥饿后细胞增殖、迁移能力的改变;对SW480细胞正常血清培养株和血清饥饿细胞亚株进行转录组测序,并对差异基因进行GO功能、KEGG通路富集分析;利用String数据库和Metascape软件构建迁移相关核心蛋白关联网络,并选取16个核心基因进行RT-qPCR验证;检测ITGB1及其相关通路关键分子在蛋白水平的表达;敲降ITGB1和使用STAT3抑制剂后利用Transwell实验检测血清饥饿组和对照组SW480细胞迁移能力的变化。结果长期血清饥饿减弱了SW480细胞的增殖能力,但增强其迁移能力;而对DLD-1细胞的增殖和迁移能力均产生抑制作用。SW480细胞转录组数据分析发现长期血清饥饿诱导细胞中3016个基因表达上调,其中迁移相关差异基因有283个;Metascape 分析发现ITGB1、CD44、TNS1、STAT3等潜在核心基因相 互关联;经验证,长期血清饥饿导致SW480细胞中VTN、 TNS1、VEGFA、STAT3、ITGB1等基因的mRNA水平显著上 调,同时导致ITGB1、MMP2等的蛋白水平和JAK2、STAT3 的磷酸化水平显著上调;敲降ITGB1和使用STAT3抑制剂 均能减弱长期血清饥饿SW480细胞的迁移能力。结论 结 直肠癌细胞可耐受慢性饥饿应激,并且该种应激可通过上 调ITGB1表达促进结直肠癌细胞的迁移能力。Objective To investigate the effects of chronic starvation stress on the proliferation and migration of colorectal cancer cells,as well as the underlying mechanisms.Methods By using prolonged serum starvation to simulate chronic starvation stress in tumor cells,we established enduring serum-deprived models of SW480 and DLD-1 cells and observed cellular morphological change.Effects of prolonged serum starvation on SW480 and DLD-1 proliferative and migratory capabilities were assessed using CCK-8 and Transwell assays.Differential gene-expression analysis on SW480 cultured with 1%FBS or 10%FBS medium was followed by GO and KEGG pathway assessments.Migration-related protein interactions were explored using String database and Metascape software,leading to 16 genes being selected for RT-qPCR validation.Protein levels of ITGB1 and key molecules in the relevant pathways were measured.Mobility changes in SW480 were observed through Transwell assay after ITGB1 knockdown or STAT3 inhibition.Results Prolonged serum starvation significantly inhibited the proliferation of SW480 and DLD-1 cells,and DLD-1 mobility,while enhanced SW480 migration.Transcriptome analysis revealed that prolonged serum deprivation caused the upregulation of 3016 genes,among which 283 were involved in cell migration.Metascape analysis identified the correlations among potential core genes ITGB1,CD44,TNS1,STAT3,etc.Prolonged serum deprivation increased the mRNA levels of VTN,TNS1,VEGFA,STAT3,and ITGB1 while also increasing the protein levels of ITGB1 and MMP2 and the phosphorylation levels of JAK2 and STAT3.Mobility reduction in prolonged serum-starved SW480 cells was achieved through ITGB1 knockdown or a STAT3 inhibitor.Conclusion Colorectal cancer cells can endure chronic starvation stress which enhances migration capability by upregulating ITGB1 expression.

关 键 词：饥饿 结直肠瘤 细胞运动 整合素Β1 STAT3转录因子 

分 类 号：R735.35[医药卫生—肿瘤] R735.37[医药卫生—临床医学]