低氧微环境增强MACC1调控PI3K/AKT信号通路促进结直肠癌肿瘤干细胞样特性  

Hypoxia microenvironment enhances MACC1 regulation of PI3K/AKT signaling pathway and promotes colon cancer stem cell like characteristics

作　　者：吴共发 曾宇婷 刘钰君[1] 姚雨江 邱丽浈 WU Gongfa;ZENG Yuting;LIU Yujun;YAO Yujiang;QIU Lizhen(Department of Pathology,the Fourth Affiliated Hospital of Guangzhou Medical University,Guangzhou 511300,Guangdong,China;Health Management Center,the Fourth Affiliated Hospital of Guangzhou Medical University,Guangzhou 511300,Guangdong,China)

机构地区：[1]广州医科大学附属第四医院病理科,广东广州511300 [2]广州医科大学附属第四医院健康管理中心,广东广州511300

出　　处：《中南医学科学杂志》2024年第1期51-55,共5页Medical Science Journal of Central South China

基　　金：广东省医学科研基金面上项目(B2021018);广州市科技计划项目(202102080560,202102080542,202002030450)。

摘　　要：目的探讨低氧微环境和结肠癌转移相关基因1(MACC1)对结直肠癌(CRC)肿瘤干细胞(CSC)样特性生物学行为的影响及机制。方法将空载体(LV-ctrl组)、MACC1过表达载体(LV-MACC1)转染结肠癌细胞HCT116,同时在低氧微环境条件下培养LV-MACC1细胞(LV-MACC1+hypoxia组)。分别应用CCK-8法及免疫细胞化学法、划痕实验、Transwell侵袭实验、肿瘤球形成实验检测细胞增殖、迁移能力、侵袭能力及体外肿瘤形成能力。免疫印迹法检测MACC1、CD133、CD44、AKT和p-AKT蛋白的表达,740 Y-P验证PI3K/AKT在MACC1诱导的肿瘤干细胞样特性中的作用。结果与LV-ctrl组比较,LV-MACC1组的细胞增殖、迁移和侵袭能力均增强,肿瘤球形成增多,CD44、CD133和p-AKT蛋白的表达水平增高(P<0.05)。与LV-MACC1组比较,LV-MACC1+hypoxia组的细胞增殖、迁移和侵袭能力增强,肿瘤球形成增多(P<0.05);CD44、CD133和p-AKT蛋白的表达水平增高(P<0.05)。与对照组比较,740 Y-P增加了细胞CD133和CD44蛋白表达水平(P<0.05)。结论低氧微环境增强MACC1调控PI3K/AKT信号通路促进结直肠癌细胞出现肿瘤干细胞样特性。Aim To investigate the effects and mechanism of hypoxia microenvironment and metastasis-associated colon cancer 1(MACC1)on the biological behavior of colorectal cancer(CRC)tumor stem cell(CSC)like properties.Methods Colon cancer cells HCT116 were transfected with empty vector(LV ctrl group)and MACC1 overexpression vector(LV-MACC1 group),and LV-MACC1 cells(LV-MACC1+Hypoxia group)were cultured under hypoxic microenvironment conditions.CCK-8 assay and immunocytochemistry,wound healing assay,transwell invasion experiment,and tumor sphere formation assay,were used to measure cell proliferation,migration,invasion abilities and tumor formation capacity of cancer cells,respectively.Western blotting test was used to detect expression of difference of protein level of MACC1,CD133,CD44,AKT and p-AKT.To determine the possible roles of PI3K/AKT in MACC1-induced stem cell properties,740 Y-P was applied.Results Compared with the LV-ctrl group,the cell proliferation,migration and invasion ability of LV-MACC1 group,the formation of spheres,and the expression level of CD44,CD133 and p-AKT proteins in LV-MACC1 group,were all increased(P<0.05).Compared with the LV-MACC1 group,the cell proliferation,migration and invasion ability of LV-MACC1+hypoxia group,the formation of spheres(P<0.05),and the expression level of CD44,CD133 and p-AKT protein,were all increased(P<0.05).Protein expression levels of CD133 and CD44 were increased in response of HCT116 cells to 740 Y-P,compared with in the control group(P<0.05).Conclusion Hypoxia microenvironment enhanced the ability of MACC1 to facilitate cancer stem cell like properties and malignant biological behavior in colorectal cancer cells through the PI3K/AKT signaling pathway.

关 键 词：结直肠癌 肿瘤干细胞 结肠癌转移相关基因1 低氧微环境 

分 类 号：R735.34[医药卫生—肿瘤]