结肠癌相关性调节性B细胞介导淋巴管生成的体外研究

目的：通过共培养模型，验证结肠癌相关性Breg细胞促进淋巴管生成的作用及相关机制。方法：利用结肠癌细胞CT26及MC38与B细胞构建共培养模型，诱导结肠癌相关性Breg细胞，流式细胞术鉴定CD19⁺IL-10⁺Breg细胞比例。ELISA法检测结肠癌相关性Breg细胞条件培养基中VEGF-A、VEGF-C和VEGF-D的分泌水平。利用划痕实验及管腔形成实验检测Breg条件培养基干预后淋巴结内皮细胞SVEC4-10的迁移及管腔形成能力。结果：与24h共培养体系相比，Breg^CT26诱导组(4.90±0.05)%vs.(27.63±1.12)%]和Breg^MC38诱导组(5.85±0.86)%vs.(24.27±2.27)%]在48 h共培养体系中能诱导更多的Breg细胞，且结肠癌相关性Breg诱导组VEGF-A和VEGF-C的表达水平与空白组相比显著增加(P<0.05)。管腔形成实验及划痕实验结果表明，结肠癌相关性Breg细胞能增强SVEC4-10的成管和迁移能力。结论：在肿瘤微环境中，结肠...

In vitro experimental study on lymphangiogenesis mediated by colorectal cancer-associated regulatory B cells

  Objective  To investigate the role and mechanism of action of colorectal cancer-associated regulatory B cells (Breg) in promoting lymphangiogenesis using a coculture model.   Methods  The colorectal cancer cells, CT26 and MC38, were cocultured with B cells to induce colorectal cancer-associated Breg cells; the proportion of CD19+IL-10+Breg cells was determined using flow cytometry; the secretion levels of VEGF-A, VEGF-C, and VEGF-D in conditioned medium of colorectal cancer-associated Breg cells were detected using ELISA; scratch and tube formation assays were used to detect the migration and tube formation abilities of the lymph node endothelial cell line, SVEC4-10, after intervention of Breg-conditioned medium.   Results  BregCT26-induced groups (4.90±0.05) vs. (27.63±1.12)] and BregMC38-induced groups (5.85±0.86) vs. (24.27±2.27)] from the 48h coculture system induced more Breg cells than those from the 24h coculture system; the expression levels of VEGF-A and VEGF-C in colorectal cancer-associated Breg cells were significantly higher than those in cells from the blank group (P<0.05). Moreover, colorectal cancer-associated Breg cells enhanced both the tube formation and migration abilities of SVEC4-10 cells.  Conclusions  In the tumor microenvironment, colorectal cancer cells induce B cell transformation into Breg cells. This change upregulates the secretion levels of VEGF-A and VEGF-C, enhances the tube formation and migration abilities of SVEC4-10, and promotes tumor lymphangiogenesis.