CD44抗体对卵巢癌实体瘤源球形体形成细胞增殖的抑制作用

The Inhibitory Effects and Mechanisms of Monoclonal Antibody against CD44 (A3D8) on Proliferation in Sphere-Forming Cells from Solid Tumor of Ovarian Cancer

目的:观察抗CD44单克隆抗体A3D8对卵巢癌实体瘤源球形体形成细胞增殖的作用,并初步探讨其作用机制。方法:用MTS法检测A3D8及顺铂(DDP)对细胞增殖的影响;采用流式细胞仪法观察A3D8对卵巢癌实体瘤源球形体形成细胞细胞周期的影响;彗星电泳法检测DNA损伤;通过Western blot法,进一步探讨A3D8作用于卵巢癌实体瘤源球形体形成细胞后CDK2、cyclin A蛋白表达水平的变化。结果:A3D8作用于卵巢癌实体瘤源球形体形成细胞后,细胞抑制作用明显增强(P<0.05),而且随着A3D8质量浓度的增加和时间的延长,抑制作用明显增强,与DDP联用,抑制率较单独用DDP增高;A3D8可将细胞周期阻滞于S期;A3D8作用于卵巢癌实体瘤源球形体形成细胞24h后可引起DNA损伤;A3D8组CDK2、cyclin A蛋白表达水平明显降低。结论:A3D8可能通过阻滞细胞周期发挥抑制卵巢癌实体瘤源球形体形成细胞增殖的作用。

Objective： To explore the effects and mechanisms of monoclonal antibody against CD44 （A3D8） on proliferation in sphere-forming cells from solid tumor of ovarian cancer. Methods： The MTS assay was used to evaluate the cell proliferation.The flow cytometry was used to analyze the cell cycle distribution after treatment with A3D8 and cisplatin （DDP）. And the comet assay was used to measure DNA damage. The expressions of CDK2 and cyclinA proteins were evaluated by western blot assay. Results： A3D8 inhibited the proliferation of sphere-forming cells from solid tumor of ovarian cancer; the inhibition was in concentration-and time-dependant manner （P 0.05）. Compared with DDPtreatment alone, the inhibition rate caused by A3D8 combined with DDP was increased significantly. The percentage of cells in S phase significantly increased after treatment with A3D8. DNA was damaged after treatment with A3D8 for 24 h. The western blot analysis demonstrated that the expressions of CDK2 and cyclinA proteins were suppressed significantly in A3D8 treatment group. Conclusion： A3D8 could inhibit the proliferation of sphere-forming cells from solid tumor of ovarian cancer. The inhibiting effect was probably carried out by arrest of cell cycle.