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目的检测IL-10免疫黏附素和肿瘤相关巨噬细胞(TAM)对卵巢癌细胞侵袭性的影响,并研究其关系。方法通过刺激人急性单核白血病细胞(THP-1)分化为巨噬细胞,模拟体内的TAM,进行培养,取该培养基作为TAM条件培养基,并用其培养人卵巢癌细胞株ES-2细胞;采用Matrigel invasion assay观察穿过transwell小室的细胞数目,确定TAM对ES-2细胞侵袭性的影响;采用Western blot法检测TAM培养基处理后的ES-2细胞中缺氧诱导因子1α(HIF-1α)蛋白水平的变化。并通过IL-10免疫黏附素观察其对ES-2细胞HIF-1α蛋白水平及侵袭性的影响。结果加入TAM条件培养基的ES-2细胞的侵袭性明显高于正常培养基组(P<0.05);ES-2细胞中HIF-1α蛋白水平明显高于正常培养基组(P<0.05);经IL-10免疫黏附素处理后,ES-2中HIF-1α蛋白的水平及细胞侵袭性均明显下降(P<0.05)。结论 TAM可能是通过上调IL-10水平,诱导卵巢癌细胞中HIF-1α蛋白水平升高,进而促进卵巢癌细胞的侵袭和转移,IL-10免疫黏附素可以降低卵巢癌的侵袭性。这为卵巢癌的靶向/免疫治疗奠定了一定的实验基础。
Objective To investigate the effect of IL-10 immunoadhesin and tumor-associated macrophages (TAMs) on the invasiveness of ovarian cancer cells and to study their relationship. Methods TAM was induced by human acute monocytic leukemia (THP-1) cells to differentiate into macrophages and mimic TAM in vivo. The medium was used as a conditioned medium for TAM and cultured with human ovarian cancer cell line ES-2 The number of cells passing through the transwell chamber was determined by Matrigel invasion assay to determine the effect of TAM on invasiveness of ES-2 cells. Western blot was used to detect the expression of hypoxia-inducible factor-1α (HIF-1α) in TAM- 1α) protein level changes. IL-10 immunoadhesin was used to observe the effects of IL-10 on HIF-1α protein level and invasiveness in ES-2 cells. Results The invasiveness of ES-2 cells in TAM conditioned medium was significantly higher than that in normal medium (P <0.05). The protein level of HIF-1α in ES-2 cells was significantly higher than that in normal medium (P <0.05). After IL-10 immunoadhesin treatment, the level of HIF-1α protein and cell invasiveness in ES-2 cells were significantly decreased (P <0.05). Conclusion TAM may increase the level of HIF-1α protein in ovarian cancer cells by up-regulating the level of IL-10, and then promote the invasion and metastasis of ovarian cancer cells. IL-10 immunoadhesin may reduce the invasiveness of ovarian cancer. This has laid a certain experimental foundation for the targeted / immunotherapy of ovarian cancer.