3D Culture Represents Apoptosis Induced by Trastuzumab Better than 2D Monolayer Culture
3D Culture Represents Apoptosis Induced by Trastuzumab Better than 2D Monolayer Culture
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DOI：
10.21873/anticanres.12528
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发表时间：
2018-05-01
影响因子：
2
通讯作者：
Minami, Hironobu
中科院分区：
文献类型：
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作者：
Tatara, Takashi;Mukohara, Toru;Minami, Hironobu
背景：我们的假设是，三维（3D）培养比二维（2D）培养更能代表pik3ca野生型（wt）和突变型（mt）细胞系对曲妥珠单抗的体内反应差异。材料和方法：使用western blot分析4株人表皮生长因子受体2 （HER2）扩增的乳腺癌细胞系，评估细胞凋亡和细胞信号蛋白对曲妥珠单抗有和没有BKM120（一种泛磷脂酰肌醇3-激酶（PI3K）抑制剂）的反应。结果：在曲妥珠单抗的作用下，仅在3d培养的PIK3CA-wt细胞系中观察到cleaved poly （adp -核糖）聚合酶（PARP）的表达增加，而在3d培养的PIK3CA-wt或PIK3CA-mt细胞系中则没有。在3D培养中，PIK3CA-wt细胞中磷酸化（p-）AKT / AKT的比例比PIK3CA-mt细胞更明显，而在2D培养中，PIK3CA基因型之间的差异不太明显。与单独治疗相比，BKM120和曲妥珠单抗治疗导致裂解性PARP的增加更强。结论：3D培养似乎更好地代表曲妥珠单抗诱导的细胞凋亡和对曲妥珠单抗的耐药与PIK3CA突变相关。
Background: Our hypothesis was that three-dimensional (3D) culture better represents differential in vivo responses to trastuzumab between PIK3CA-wild-type (wt) and mutant (mt) cell lines than does two-dimensional (2D) culture. Materials and Methods: Apoptosis and cell signaling proteins were evaluated in response to trastuzumab with and without BKM120, a pan-phosphatidylinositol 3-kinase (PI3K) inhibitor, using western blot analysis of four breast cancer cell lines with human epidermal growth factor receptor 2 (HER2) amplification. Results: Increased expression of cleaved poly (ADP-ribose) polymerase (PARP) was observed only in 3D-cultured PIK3CA-wt lines in response to trastuzumab, but not in 2D-cultured PIK3CA-wt or PIK3CA-mt lines. Decrease in the ratio of phosphorylated (p-)AKT to AKT in response to trastuzumab was more profound in PIK3CA-wt cells than in PIK3CA-mt cells in 3D culture, while the difference between PIK3CA genotypes was less apparent in 2D culture. Treatment with BKM120 and trastuzumab resulted in a stronger increase in cleaved PARP than either treatment alone. Conclusion: 3D Culture appears to better represent trastuzumab-induced apoptosis and resistance to trastuzumab associated with PIK3CA mutation.