改良的人乳腺癌细胞三维培养模型及药敏性考察

An Improved Three-dimensional Cell Culture Model of Human Breast Cancer as Well as Drug Sensitivity Investigation

目的:建立适用于高通量药物筛选的人乳腺癌MCF-7细胞三维培养模型,考察其药敏性。方法:在传统的liquidoverlay法和直接混合法基础上,建立新型锚定法,制备MCF-7细胞的三维培养系统;以二维培养和直接混合法制备的三维培养系统作对照,通过倒置显微镜观察其中细胞形态学特征,绘制细胞生长曲线,使用顺铂并采用MTT法进行药敏试验。结果:采用新型锚定法成功制备MCF-7细胞的三维培养系统,不同于二维培养系统,其中MCF-7细胞形成典型的直径为160μm左右的葡萄样多细胞球;与直接混合法制备的三维培养系统相比,其中MCF-7细胞生长速度明显更快(P<0.05);与二维培养系统相比,其中MCF-7细胞对顺铂的敏感性显著降低,即顺铂对MCF-7细胞的增殖抑制率明显降低(P<0.01)。结论:该新型锚定法制备的乳腺癌细胞三维培养系统更接近体内肿瘤生长环境,其中肿瘤细胞能模拟体内实体瘤的耐药特征,故其更适用于抗癌药物的活性筛选。

Objective： To establish a three-dimensional （sO） culture model of human breast cancer cell line MCF-7 for high throughout screening of anticancer agents and to investigate its drug sensitivity.Methods： Based on the conventional liquid overlay method and mixing method, the improved anchor method was established for the preparation of a new sO culture system of MCF-7 cell line. Using a 20 culture system and a sO culture system prepared by mixing method as control, the morphological characteristics of MCF-7 cells in the new sO culture system were observed under inverted microscope, the cell growth curve was plotted and the drug sensitivity test was conducted with OOP by MTT method. Results： The new 3D culture system of MCF-7 cell line was successfully established by the improved anchor method. The typical grape-like multicellular spheroids （-160μm） were observed in the new 3D system rather than in the 20 culture system. The cell growth rate in the new 3D culture system was significandy faster than that in the conventional 3D culture system prepared by mixing method （P〈0.05）. The cell proliferation inhibition ratio by ODP in the new 3D system was significandy lowered compared with that in the 2D culture system （P〈0.01）. Conclusion：The new 3D cell culture system of human breast cancer prepared by the improved anchor method is close to in viw tumor growth environment, in which the tumor cells could mimic the drug-resistant characteristics of in viw solid tumor. Therefore the new 3D cell culture system is more practicable for the activity screening of anticancer candidates than the conventional ones.