Discoidin domain receptor 2 deficiency predisposes hepatic tissue to colon carcinoma metastasis
Discoidin domain receptor 2 deficiency predisposes hepatic tissue to colon carcinoma metastasis
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DOI：
10.1136/gutjnl-2011-300810
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发表时间：
2012-10-01
期刊：
影响因子：
24.5
通讯作者：
Vidal-Vanaclocha, Fernando
中科院分区：
文献类型：
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作者：
Badiola, Iker;Olaso, Elvira;Vidal-Vanaclocha, Fernando
背景 肝星状细胞（HSC）向肌成纤维细胞的转分化是肝转移中基质发育的主要机制，但其调控途径仍不清楚。纤维化肝脏转分化的 HSC 表达高水平的纤维状胶原受体盘状结构域受体 2 (DDR2)，但尚不清楚 DDR2 是否在肿瘤微环境中发挥直接促纤维化作用。目的 评估 DDR2 对 HSC 来源的肌成纤维细胞促转移作用的影响。方法通过脾内注射在 DDR2(-/-) 和 DDR2(+/-) 和 DDR2(+/-) 小鼠中诱导肝转移注射 MCA38 结肠癌细胞，并表征其生长和特征。通过免疫组织化学对转移灶中的基质生成、血管生成和癌细胞增殖反应进行定量。在原代培养的肝窦内皮细胞 (LSEC) 和 MCA38 细胞中评估了在 MCA38 细胞条件培养基中孵育的原代培养的 DDR2(-/-) 和 DDR2(+/+) HSC 上清液的粘附、迁移和增殖刺激活性。比较了新鲜分离的 DDR2(-/-) 和 DDR2(+/+) HSC 的基因表达特征，并在基础条件下和用 MCA38 肿瘤条件培养基刺激后通过 RT-PCR 研究了 DDR2 调节基因。 结果 DDR2(-/-) 肝脏中的转移灶增加了三倍，并且包含更高密度的表达 α-平滑肌肌动蛋白的肌成纤维细胞、表达 CD31微血管和表达 Ki67 的 MCA38 细胞比 DDR2(+/+) 肝脏中的转移灶更明显。与 DDR2(+/+) HSC 相比，MCA38 激活的 DDR2(-/-) HSC 条件培养基显着增加了 LSEC 和 MCA38 细胞的粘附、迁移和增殖。 HSC 中 DDR2 缺陷导致干扰素 γ 诱导因子白细胞介素 (IL)-18 和胰岛素样生长因子-I 的基因表达降低；促转移因子 IL-10、转化生长因子 (TGF)β 和血管内皮生长因子 (VEGF)、骨形态发生蛋白 7 和 syndecan-1 的基因表达增加。 MC38肿瘤条件培养基进一步加剧了DDR2依赖性IL-10、TGFβ和VEGF基因的表达变化。结论DDR2缺陷促进肿瘤激活的HSC的肌成纤维细胞转分化，通过HSC衍生因子在肝脏中产生促转移微环境。这些发现强调了基质细胞通过改变受体-基质相互作用来调节肝脏微环境以促进转移的作用。
Background The transdifferentiation of hepatic stellate cells (HSCs) into myofibroblasts is a major mechanism for stroma development in hepatic metastasis, but their regulatory pathways remain unclear. Transdifferentiated HSCs from fibrotic liver express high levels of the fibrillar collagen receptor discoidin domain receptor 2 (DDR2), but it is unclear if DDR2 plays a direct profibrogenic role in the tumour microenvironment.Aim To assess the impact of DDR2 on the prometastatic role of HSC-derived myofibroblasts.Methods Hepatic metastases were induced in DDR2(-/-) and DDR2(+/+) mice by intrasplenic injection of MCA38 colon carcinoma cells, and their growth and features were characterised. Stromagenic, angiogenic and cancer cell proliferation responses were quantified in metastases by immunohistochemistry. The adhesion-, migration-and proliferation-stimulating activities of supernatants from primary cultured DDR2(-/-) and DDR2(+/+) HSCs, incubated in MCA38 cell-conditioned medium, were evaluated in primary cultured liver sinusoidal endothelium cells (LSECs) and MCA38 cells. Gene expression signatures from freshly isolated DDR2(-/-) and DDR2(+/+) HSCs were compared and DDR2-regulated genes were studied by RT-PCR under basal conditions and after stimulation with MCA38 tumour-conditioned media.Results Metastases were increased three fold in DDR2(-/-) livers, and contained a higher density of alpha-smooth muscle actin-expressing myofibroblasts, CD31-expressing microvessels and Ki67-expressing MCA38 cells than metastases in DDR2(+/+) livers. Media conditioned by MCA38-activated DDR2(-/-) HSCs significantly increased adhesion, migration and proliferation of LSECs and MCA38 cells, compared with DDR2(+/+) HSCs. DDR2 deficiency in HSCs led to decreased gene expression of interferon gamma-inducing factor interleukin (IL)-18 and insulin-like growth factor-I; and increased gene expression of prometastatic factors IL-10, transforming growth factor (TGF)beta and vascular endothelial growth factor (VEGF), bone morphogenetic protein-7 and syndecan-1. MC38 tumour-conditioned media further exacerbated expression changes in DDR2-dependent IL-10, TGF beta and VEGF genes.Conclusion DDR2 deficiency fosters the myofibroblast transdifferentiation of tumour-activated HSCs, generating a prometastatic microenvironment in the liver via HSC-derived factors. These findings underscore the role of stromal cells in conditioning the hepatic microenvironment for metastases through altered receptor-stroma interactions.