Human Colon Cancer Derived Mesenchymal Stem Cells Promote the Tumorigenicity of Colon Cancer by Secreting Exosomes
LIU Hong-yue1,2, YU Bo3, ZHAI Qing1,2,4*
1. Department of Pharmacy, Fudan University Shanghai Cancer Center, Shanghai 200032, China; 2. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; 3. Tongren Hospital, School of Medicine,Shanghai Jiaotong University, Shanghai 200336, China; 4. School of Pharmacy, Fudan University, Shanghai 201203, China
摘要目的 探讨人结肠癌来源的间充质干细胞所分泌的外泌体对结肠癌细胞生物学功能的影响。方法 将人结肠癌干细胞诱导培养出人结肠癌间充质干细胞(human colon cancer mesenchymal stem cells,hCC-MSCs),利用流式细胞学和成骨、成脂诱导分化实验验证间充质干细胞的多能性。利用ExoQuick试剂盒提取hCC-MSCs外泌体(exosomes secreted from human colon cancer mesenchymal stem cells,hCC-MSCs-Exo),通过电镜、粒径分析和Western blot进行鉴定。采用PKH67标记hCC-MSCs-Exo,与HCT-116和HT-29共培养,荧光显微镜观察结肠癌细胞株摄取外泌体的情况,最后利用CCK8增殖实验、细胞划痕实验和Transwell侵袭实验检测hCC-MSCs-Exo对结肠癌细胞株生物学功能的影响。结果 hCC-MSCs细胞呈单层贴壁生长,阳性表达CD29、CD90及CD105,阴性表达CD34、CD45及HLA-DR,具有成骨分化和成脂分化的多能性。hCC-MSCs-Exo平均直径135 nm,阳性表达Alix、CD63及CD81。hCC-MSCs-Exo与结肠癌细胞株共培养后,结肠癌细胞株能够摄取hCC-MSCs-Exo,且主要分布于细胞质中。CCK8、划痕实验和Transwell实验表明,与对照组相比,共培养组结肠癌细胞的增殖能力、迁移能力和侵袭能力显著增强(P<0.05)。结论 hCC-MSCs-Exo能够促进结肠癌细胞增殖、迁徙和侵袭,为结肠癌药物治疗提供潜在的新治疗靶点。
Abstract：OBJECTIVE To explore the effect of exosome from human colon cancer derived mesenchymal stem cells on the biological function of colon cancer cells. METHODS Human colon cancer mesenchymal stem cells (hCC-MSCs) were induced from human colon cancer stem cells, and verified by flowcytometry, osteogenesis and adipogenesis differentiation. The exosomes were purified by ExoQuick Exosome Precipitation kit and characterized by electron microscopy, nanoparticle tracking analysis and Western blot. PKH67-labeled hCC-MSCs-Exo was co-cultured with HCT-116 and HT-29. The effects of hCC-MSCs-Exo on the biological function of colon cancer cell lines were investigated by CCK8 assay, wound healing assay and Transwell invasion assay. RESULTS The hCC-MSCs showed monolayer adherent growth, fibrocyte-like with positive expression of CD29, CD90, CD105 and negative expression of CD34, CD45, HLA-DR, which have the abilities of osteoblasts and adipocytes. hCC-MSCs-Exo has an average diameter of 135 nanometers and positive expression of Alix, CD63 and CD81. After co-culture with colon cancer cell lines, hCC-MSCs-Exo was uptaken into the colon cancer cells. CCK8, wound healing and Transwell assays showed that the proliferation, migration and invasion ability of co-culture group were significantly enhanced compared with control group (P<0.05). CONCLUSION hCC-MSCs-Exo can promote the proliferation, migration and invasion of colon cancer cells. It is expected to be a new target for colon cancer treatment.
刘洪悦, 余波, 翟青. 人结肠癌来源的间充质干细胞通过分泌外泌体促进结肠癌发生发展的机制研究[J]. 中国药学杂志, 2020, 55(12): 1008-1014.
LIU Hong-yue, YU Bo, ZHAI Qing. Human Colon Cancer Derived Mesenchymal Stem Cells Promote the Tumorigenicity of Colon Cancer by Secreting Exosomes. Chinese Pharmaceutical Journal, 2020, 55(12): 1008-1014.
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2020, Vol. 55 
