CAJ | 학술논문

目的:探讨抗CD47单克隆抗体在体外对巨噬细胞吞噬卵巢癌细胞作用的影响.方法:收集天津医科大学附属肿瘤医院2011年6月至2011年12月手术切除的卵巢组织样本45例(包含卵巢癌组织样本39例,正常卵巢组织样本6例),获得的组织细胞通过流式细胞术分析CD47的表达水平,通过细胞功能学实验观察抗CD47单克隆抗体在体外对小鼠巨噬细胞吞噬卵巢癌细胞的靶向治疗作用.结果:流式细胞术分析显示,卵巢癌细胞系SKOV-3细胞和卵巢癌组织细胞中CD47表达水平均明显高于正常人卵巢上皮细胞,差异具有统计学意义(P<0.01),且与疾病的分化程度相关(P<0.05).细胞功能学实验结果显示,抗CD47单克隆抗体在体外可促进巨噬细胞对卵巢癌细胞的吞噬作用,各实验组(1μg/mL、5μg/mL、10μg/mL、20μg/mL)分别较空白对照组吞噬指数显著增加,并且随抗体浓度的增加吞噬指数呈浓度依赖性,差异均具有统计学意义(P<0.01).统计学分析显示,10μg/mL浓度组吞噬指数达到峰值,同20μg/mL浓度组吞噬指数无显著性差异(P>0.05).结论:CD47高表达于卵巢癌组织细胞,且与组织分化程度相关.抗CD47单克隆抗体在体外可促进巨噬细胞对卵巢癌肿瘤细胞的吞噬作用,为卵巢癌的靶向治疗提供了实验依据.
목적:탐토항CD47단극륭항체재체외대거서세포탄서란소암세포작용적영향.방법:수집천진의과대학부속종류의원2011년6월지2011년12월수술절제적란소조직양본45례(포함란소암조직양본39례,정상란소조직양본6례),획득적조직세포통과류식세포술분석CD47적표체수평,통과세포공능학실험관찰항CD47단극륭항체재체외대소서거서세포탄서란소암세포적파향치료작용.결과:류식세포술분석현시,란소암세포계SKOV-3세포화란소암조직세포중CD47표체수평균명현고우정상인란소상피세포,차이구유통계학의의(P<0.01),차여질병적분화정도상관(P<0.05).세포공능학실험결과현시,항CD47단극륭항체재체외가촉진거서세포대란소암세포적탄서작용,각실험조(1μg/mL、5μg/mL、10μg/mL、20μg/mL)분별교공백대조조탄서지수현저증가,병차수항체농도적증가탄서지수정농도의뢰성,차이균구유통계학의의(P<0.01).통계학분석현시,10μg/mL농도조탄서지수체도봉치,동20μg/mL농도조탄서지수무현저성차이(P>0.05).결론:CD47고표체우란소암조직세포,차여조직분화정도상관.항CD47단극륭항체재체외가촉진거서세포대란소암종류세포적탄서작용,위란소암적파향치료제공료실험의거.
Objective:CD47 is highly expressed on human ovarian cancer cell surface and is a ligand for signal-regulatory pro-tein alpha (SIRPα). The CD47-SIRPαpathway has been found to negatively regulate macrophage phagocytosis. We hypothesize that this pathway enables ovarian cancer cells to escape during tumor immunosurveillance. CD47 is highly expressed in various malignant diseases. This study aimed to explore whether CD47 can serve as a potential therapeutic target for ovarian cancer. Methods:Tissue sam-ples were obtained from 39 epithelial ovarian cancer patients. Six patients underwent other operations during which normal ovarian tis-sue samples were obtained. The expression of CD47 in the ovary cancer cell line SKOV-3 in normal ovarian tissues and ovarian cancer cells were analyzed by quantitative fluorescence-activated cell sorting. Macrophages and CFSE-labeled SKOV-3 cells were co-cultured with the indicated antibodies through in vitro phagocytosis assay, and phagocytic index was calculated. Results: Flow cytometry showed that the expression levels of CD47 in the ovarian cancer cell line SKOV-3 and ovarian cancer cells were significantly higher than in normal ovarian epithelial cells (P<0.01). This result is associated with the grade of differentiation of the disease (P<0.05). Block-ing a monoclonal antibody against CD47 promotes in vitro phagocytosis of ovarian cancer cells by macrophages. The phagocytic index of the experimental groups (1, 5, 10, and 20μg/ml) increased significantly (P<0.01) compared with the control group. The occurrence was found to be concentration-dependent (P<0.01). Statistical analysis showed that the maximum phagocytic index was found in the 10μg/ml concentration group;the same was observed in the 20μg/ml group. However, no significant difference was found between the two groups (P>0.05). Conclusion:CD47 is expressed in epithelial ovarian cancer cells at a higher level compared with normal ovarian cells. The level of expression is associated with tissue differentiation grade. Our study indicates that the anti-CD47 monoclonal anti-body can significantly promote in vitro phagocytosis, indicating that the anti-CD47 monoclonal antibody might be a potential target in targeted cancer therapy.