SHCBP1通过调控PI3K/AKT/mTOR信号通路促进结直肠癌Caco-2细胞增殖、迁移和侵袭OA
SHCBP1 promotes proliferation,migration,and invasion of colorectal cancer Caco-2 cells by regulating the PI3K/AKT/mTOR signaling pathway
目的:探讨Shc SH2结构域结合蛋白1(SHCBP1)基因在结直肠癌(CRC)中的表达及其调控CRC细胞(Caco-2)增殖、迁移和侵袭的机制.方法:将Caco-2细胞分为4个实验组:Con组(空白对照)、sh-NC组(转染阴性对照sh-NC)、sh-SHCBP1组(转染sh-SHCBP1)、sh-SHCBP1+740Y-P组(转染sh-SHCBP1 后用 30 μmol/L PI3K激活剂740Y-P处理 1 h).采用 CCK-8法、细胞划痕和Transwell实验检测细胞增殖、迁移和侵袭能力,基于Matrigel进行三维培养实验检测SHCBP1对Caco-2细胞血管生成拟态(VM)形成能力的影响,qRT-PCR检测SHCBP1的mRNA表达情况,WB法检测SHCBP1、VM相关蛋白以及PI3K/AKT/mTOR信号通路相关蛋白的表达.结果:SHCBP 1在CRC组织和细胞中呈高表达(P＜0.05).与Con组和sh-NC组相比,sh-SHCBP1组的SHCBP 1的mRNA和蛋白表达水平、细胞增殖活性、划痕修复率、穿膜细胞数和形成的小管数均显著降低(均P＜0.05);低氧诱导因子1α(HIF-1α)、EphrinA型受体2(EPHA2)、VEGFA、p-PI3K、p-AKT和p-mTOR蛋白表达水平均显著降低(均P＜0.05).对比sh-SHCBP1组,sh-SHCBP1+740Y-P组的细胞增殖活性、划痕修复率、穿膜细胞数和形成的拟态血管数均显著增加(均P＜0.05);HIF-1α、EPHA2、VEGFA、p-PI3K、p-AKT和p-mTOR蛋白表达水平均显著增加(均P＜0.05).结论:SHCBP 1在Caco-2细胞中表达显著上调,促进Caco-2细胞的增殖、迁移和侵袭,其机制可能与激活PI3K/AKT/mTOR信号通路促进VM过程有关.
Objective:To investigate the expression of Shc SH2 domain-binding protein 1(SHCBP1)in colorectal cancer(CRC)and its regulatory mechanism on Caco-2 cell proliferation,migration,and invasion.Methods:Caco-2 cells were divided into four experimental groups:the Con group(blank control),the sh-NC group(transfection with negative control sh-NC),the sh-SHCBP1 group(transfection with sh-SHCBP1),and the sh-SHCBP1+740Y-P group(transfection with sh-SHCBP1 followed by treatment with the PI3K activator 740Y-P at 30 μmol/L for 1 h).Cell proliferation,migration,and invasion were assessed using the CCK-8 assay,wound-healing,and Transwell invasion assay,respectively.Vasculogenic mimicry(VM)formation in Caco-2 cells was examined using a Matrigel-based three-dimensional culture system.The mRNA expression level of SHCBP1 was detected using qRT-PCR,and the protein expression levels of SHCBP1,VM-related markers,and PI3K/AKT/mTOR signaling pathway-related proteins were detected using WB.Results:SHCBP1 was significantly upregulated in CRC tissues and cell lines(P＜0.05).Compared with Con group and sh-NC group,the sh-SHCBP1 group showed significantly reduced mRNA and protein levels of SHCBP1,cell proliferation activity,wound-healing rate,number of invaded cells,and number of formed tubules(all P＜0.05).In addition,the protein expression levels of hypoxia-inducible factor 1α(HIF-1α),ephrin type-A receptor 2(EPHA2),VEGFA,p-PI3K,p-AKT,and phosphorylated mTOR(p-mTOR)were significantly decreased(all P＜0.05).Compared with the sh-SHCBP1 group,the sh-SHCBP1+740Y-P group demonstrated significantly increased cell proliferation,wound-healing rate,number of invaded cells,and number of formed VM structures(all P＜0.05),along with significantly elevated protein expression of HIF-1α,EPHA2,VEGFA,p-PI3K,p-AKT,and p-mTOR(all P＜0.05).Conclusion:SHCBP1 is highly expressed in CRC cells and promotes the proliferation,migration,and invasion of Caco-2 cells.The underlying mechanism may be associated with activation of the PI3K/AKT/mTOR signaling pathway and subsequent promotion of the VM process.
李丽;姚红亮
衡水市人民医院普外二科,河北衡水 053000衡水市人民医院普外二科,河北衡水 053000
医药卫生
SHCBP 1PI3K/AKT/mTOR结直肠癌血管生成拟态Caco-2细胞
SHCBP1PI3K/AKT/mTORcolorectal cancer(CRC)vasculogenic mimicry(VM)Caco-2 cell
《中国肿瘤生物治疗杂志》 2026 (2)
174-180,7
河北省医学科学研究课题计划(20211322)
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