AggreWell板培养乳腺癌肿瘤球及苦参碱对侵袭迁移蛋白分泌的影响

AggreWell Plate Culture of Breast Cancer Tumor Spheroids and the Impact of Matrine on the Secretion of Proteins Associated with Invasion and Migration

摘要: 目的：探讨通过AggreWell^TM800板培养MDA-MB-231细胞，构建3D乳腺癌肿瘤球，并用于苦参碱影响乳腺癌MDA-MB-231细胞侵袭迁移蛋白分泌的可行性。方法：AggreWell^TM800 24孔板经处理后种植MDA-MB-231细胞，培养三维乳腺癌肿瘤球。采用ELISA法测定不同剂量苦参碱影响乳腺癌MDA-MB-231细胞3D肿瘤球培养上清中基质金属蛋白酶(MMP-2、MMP-9)和上皮间质转化(EMT)标记物E-cadherin、N-cadherin、Vimentin的表达情况。并与传统2D单层培养条件下进行比较。结果：苦参碱可有效抑制MDA-MB-231细胞2D/3D培养上清液中MMP-2、MMP-9的表达，以及有效抑制EMT，表现为下调N-cadherin、Vimentin表达，上调E-cadherin表达。随着苦参碱浓度的增加，抑制增加，呈剂量依赖性。且对3D培养的抑制显著低于对2D培养的抑制。结论：本研究表明，AggreWell构建的3D模型可能比传统2D模型更能反映乳腺癌细胞对苦参碱的耐受性，提示其在临床前药物筛选中可能具有潜在优势。

Abstract: Objective: To investigate the feasibility of generating three-dimensional (3D) breast cancer tumor spheroids using MDA-MB-231 cells cultured in AggreWell^TM800 plates, and to evaluate the effects of matrine on the secretion of invasion- and migration-related proteins in these cells. Methods: MDA-MB-231 cells were seeded into 24-well AggreWell^TM800 plates to generate three-dimensional breast cancer tumor spheroids. The expression levels of matrix metalloproteinases (MMP-2 and MMP-9) and Epithelial-Mesenchymal Transition (EMT) markers, including E-cadherin, N-cadherin, and Vimentin, were quantified via ELISA in the supernatant of the 3D tumor spheroid cultures under varying concentrations of matrine. These results were subsequently compared with those obtained under conventional two-dimensional monolayer culture conditions. Results: Matrine significantly suppressed the expression of MMP-2 and MMP-9 in both 2D and 3D culture systems in a dose-dependent manner. Furthermore, matrine inhibited EMT, as evidenced by the downregulation of N-cadherin and Vimentin protein expression and the upregulation of E-cadherin. However, the inhibitory effects observed in the 3D culture model were markedly less pronounced than those in the 2D model. Conclusion: This study demonstrates that the 3D cell model generated using AggreWell more accurately reflects the tolerance of breast cancer cells to matrine compared to the conventional 2D model, indicating its potential advantages in preclinical drug screening applications.