壳聚糖-透明质酸支架用于低氧靶向性抗癌药物筛选的研究

Chitosan-hyaluronic Acid Scaffolds for Screening Hypoxia-targeted Anticancer Drugs

三维细胞培养近年来正被逐渐广泛地应用于抗肿瘤药物筛选,与传统二维细胞培养相比,三维细胞培养模型中的细胞形态、结构和功能与实体瘤更加接近,有利于模拟肿瘤微环境,从而提高药物筛选的准确度。以壳聚糖和透明质酸为原料制备了一种三维多孔支架,并使用该支架构建了人脑胶质瘤SF763细胞的三维肿瘤模型。在此基础上,使用该三维肿瘤球和普通二维细胞评价了尼莫司汀(ACNU)与替拉扎明(TPZ)的体外抗肿瘤活性;并探讨了在常氧和低氧条件下两种药物的活性差异。结果表明,不论在常氧还是低氧条件下,经ACNU处理的三维细胞均比二维细胞表现出更强的耐药性,这说明三维细胞在抗癌药物耐药性评价中具有更高的灵敏性。对于肿瘤低氧敏感药物TPZ,其对三维细胞的毒性与低氧条件下二维细胞的毒性接近,且显著高于在常氧条件下对二维细胞的毒性,这说明三维肿瘤球模拟了肿瘤组织的低氧微环境,使TPZ发生低氧启动从而发挥了抗肿瘤活性。三维细胞培养可作为抗肿瘤药物体外活性评价的有效模型,为抗肿瘤药物筛选提供了更加可靠的方法。

The three-dimensional(3D)cell culture has been widely used for screening of anticancer agents.Compared with traditional two-dimensional(2D)cell culture,the cell morphology,structure and function of 3D tumor culture model are more similar to solid tumors,which is beneficial to simulate the tumor microenvironment and to improve the accuracy of drug screening.In this study,chitosan and hyaluronic acid(HA)were used to synthesize 3D porous scaffolds,which were applied for establishing a 3D tumor model of human brain glioma SF763 cells.Based on the 3D model,the anticancer activity of nimustine(ACNU)and tirapazamine(TPZ)were investigated in vitro using 3D tumor spheres and 2D cells.Besides,the activity of the two drugs under normoxia and hypoxic conditions was compared.The results showed that the 3D cells treated with ACNU showed higher drug resistance than the 2D cells under both normal and hypoxic conditions,indicating that 3D cells possess higher sensitivity in the evaluation of drug resistance to chemotherapies.Tirapazamine,a tumor hypoxia-sensitive drug,exhibited similar toxicity to 3D and 2D cells under hypoxic condition,which was significantly higher than that under normoxic condition.This indicated that the 3D tumor spheres simulating the hypoxic microenvironment of tumors tissue enabled the hypoxic activation of TPZ,thus exerting its anti-tumor activity.This study showed that 3D cell culture could be used as an effective model for in vitro activity evaluation of anti-tumor drugs,and provided a reliable method for screening antitumor drugs.