摘要
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 immunoadhesin for IL - 10 and tumor associated macrophages（TAM） on the invasion of ovarian cancer cells and the relationship between immunoadhesin for IL - 10 and TAM. METHODS By stimulating THP - 1 to differentiate into macrophages, which was used to imitate intracorporal TAM. The culture medium was collected to get TAM conditioned medium. The ES - 2 cells were then treated with conditioned medium. The effect of TAM on the invasion of ES - 2 cells was determined by matrigel invasion assay to observe the number of cells passing through the transwell chamber. The change of HIF - lot in ES -2 cells treated with TAM was detected by Western blot. Besides,the effect of IL - 10 antagonist on the level of HIF - lot protein and invasiveness of ES - 2 cells was observed. RESULTS The invasiveness of ES - 2 cells was significantly higher than that of the normal medium（ P 〈 0.05 ）. The level of HIF - 1a protein in ES -2 cells was also significantly higher than that in normal medium （P 〈 0.05 ）. The level of HIF - lot protein and cell invasiveness in ES -2 were significantly decreased after treatment with IL - 10 antagonist（ P 〈 0. 05 ）. CONCLUSION TAM may up - regulate the level of IL - 10, induce the elevation of HIF - Iot protein in ovarian cancer cells, and promote the invasion and metastasis of ovarian cancer cells. This has laid a certain experimental basis for targeting/immunotherapy of ovarian cancer.
出处
《华西药学杂志》
CAS
CSCD
2017年第3期257-259,共3页
West China Journal of Pharmaceutical Sciences
基金
中国癌症基金会资助项目(批准号:S2014025)
