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目的:研究重组人血管内皮抑素(商名品:恩度)是否可通过抑制结肠癌细胞EDA表达从而影响结肠癌淋巴管生成。方法:分别用单纯培养和经重组人血管内皮抑素处理的SW480细胞培养上清液处理人淋巴管内皮细胞(hLECs),三维培养观察hLECs体外成管能力的差异,蛋白质印迹法检测hLECs中integrinα9的表达差异。结果:SW480细胞高表达EDA蛋白片断,重组人血管内皮抑素呈浓度依赖性地抑制EDA片断在SW480中的表达。单纯培养的SW480培养上清液(条件培养基Ⅰ)可促进hLECs体外成管,SW480经重组人血管内皮抑素处理后其培养上清液(条件培养基Ⅱ)促hLECs体外成管的效应被削弱,hLECs小管形成数量明显减少,各实验组小管形成数量差异有统计学意义,P<0.01。与之对应地,条件培养基Ⅱ处理组中hLECsEDA受体integrinα9的表达水平较条件培养基Ⅰ中hLECs下降(P<0.05)。结论:EDA在肿瘤淋巴管生成中具有重要作用,重组人血管内皮抑素可通过EDA-integrinα9途径有效抑制hLECs体外成管能力。
OBJECTIVE: To study whether recombinant human endostatin (a brand name: Endostar) can affect the lymphangiogenesis of colon cancer by inhibiting the expression of EDA in colon cancer cells. Methods: Cultured human hepatocellular carcinoma cells (hLECs) were treated with SW480 cell culture supernatant and cultured with recombinant human endostatin respectively. The ability of hLECs in vitro was observed by three-dimensional culture. Western blotting was used to detect the expression of integrin α9 Differences in expression. Results: ED4 protein was highly expressed in SW480 cells. Recombinant human endostatin inhibited the expression of EDA fragment in SW480 in a concentration - dependent manner. The culture of SW480 culture supernatant (conditioned medium Ⅰ) can promote the in vitro tube formation of hLECs in vitro, and the effect of SW480 on the in vitro tube formation of hLECs induced by recombinant human endostatin in culture supernatant (conditioned medium Ⅱ) Weakened, hLECs tubule formation significantly reduced the number of tubules in each experimental group the number of differences was statistically significant, P <0.01. Correspondingly, the hLECsEDA receptor integrinα9 expression in conditioned medium Ⅱ group decreased compared with hLECs in conditioned medium Ⅰ (P <0.05). Conclusion: EDA plays an important role in lymphangiogenesis of tumor. Recombinant human endostatin can effectively inhibit the in vitro tube formation ability of hLECs through EDA-integrin α9 pathway.