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13:19 min
April 26th, 2024
DOI :
10.3791/65922-v
Chapters
0:12
Introduction
1:07
Setting up the JIMT-1 Enhanced Fluorescent Protein (EGFP) Spheroid Model
3:43
Coating the HCS Plate
4:33
Transfer of Spheroids to the HCS Plate
4:46
Pre-treatment of JIMT-1-EGFP Spheroids with Test Compounds
5:14
Induction of Antibody-Dependent Cellular Cytotoxicity (ADCC) by Adding the Effector Cells
7:10
Annexin V-647 Staining of Spheroids
7:34
Imaging
11:43
Representative Results
12:52
Conclusion
Transcript
最有效的靶向癌症治疗之一是基于抗体,例如抗 HER2 抗体曲妥珠单抗。曲妥珠单抗的抗癌作用涉及抗体依赖性细胞介导的细胞毒性 ADCC,通过自然杀伤细胞或 NK 细胞。了解调节癌细胞对抗生长因子受体抗体治疗敏感性的机制,特别是ADCC,对于开发新的、更有效的治疗方式可能至关重要。
癌细胞球体等3D模型在预测肿瘤对各种抗癌疗法的免疫反应方面被证明具有优越性。因此,我们使用 NK92 自然杀伤细胞系、表达 GFP 的 JIMT-1 乳腺癌细胞和曲妥珠单抗建立了 ADCC 球状体模型。
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Summary
在这里,我们提出了一种鉴定调节ADCC机制的化合物的方法,ADCC机制是抗肿瘤抗体的重要癌细胞杀伤机制。在曲妥珠单抗存在下,在乳腺癌细胞球状体中测量NK细胞的细胞毒性作用。图像分析可识别球状体中的活细胞和死细胞、杀伤细胞和靶细胞。
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