3D生物打印前列腺癌模型用于唑来膦酸药敏试验

3D bioprinted prostate cancer model and its application in drug sensitivity test of zoledronic acid

摘要:

目的 探讨3D生物打印模型用于唑来膦酸对PC-3前列腺癌细胞药敏试验的可行性。方法 使用3D生物打印技术构建PC-3前列腺癌3D模型，采用唑来膦酸进行药敏试验。使用钙黄绿素-AM/碘化丙啶溶液对前列腺癌模型进行Live/Dead染色测试支架内前列腺癌细胞存活生长情况。采用ELISA法测定培养上清液中人基质金属蛋白酶2（MMP-2）、人基质金属蛋白酶9（MMP-9）蛋白分泌变化。并均与2D培养条件下进行比较。结果 通过3D生物打印技术构建PC-3前列腺癌模型，细胞多层包埋在海藻酸钠/明胶支架内。结果表明，随着加入唑来膦酸浓度的升高，PC-3细胞存活率呈下降趋势。在3D培养条件下对PC-3前列腺癌细胞的整体药物敏感性显著低于2D培养。即与2D培养相比，随着唑来膦酸浓度的升高，3D细胞培养的整体耐药性显著增加，细胞培养上清液MMP-2和MMP-9蛋白分泌的下降速度明显更慢。结论 相比于2D培养，使用3D生物打印技术构建的前列腺癌模型，可能更好地模拟体内前列腺癌肿瘤微环境，以提高临床前药物研究的预测能力，改善药物临床转化。

Abstract:

Objective To explore the feasibility of 3D bioprinted model for drug sensitivity test of zoledronic acid to PC-3 prostate cancer cells.Methods PC-3 prostate cancer 3D model was constructed by 3D bioprinting technology, and drug sensitivity test was performed with zoledronic acid. Then Live/Dead staining was used to test the survival and growth of prostate cancer cells in the prostate cancer model using calcein-AM/propidium iodide solution, and the protein expression of MMP-2 and MMP-9 in culture supernatant was determined by ELISA. In addition,the results were compared with those under 2D culture condition.Results PC-3 prostate cancer model was constructed by 3D bioprinting technology, and cells were embedded in sodium alginate/gelatin scaffold. The results showed that, with the increase of zoledronic acid concentration, the survival rate of PC-3 cells decreased, and the overall sensitivity of the zoledronic acid to PC-3 prostate cancer cells in 3D culture was significantly lower than that in 2D culture. That is, compared with 2D culture, overall drug resistance of 3D cell culture was significantly increased. In addition, the descent rate of MMP-2 and MMP-9 protein secretion in the supernatant of 3D cell culture was significantly slower than that in the 2D culture model.Conclusion Compared with 2D culture, the prostate cancer model constructed by 3D bioprinting technology may better simulate the microenvironment of prostate cancer tumor in vivo, so as to improve the predictive ability of preclinical drug research and clinical transformation.