Tu Junfang, Luo Xun, Liu Haitao, Zhang Jifeng, He Mei
School of Biological Engineering, Huainan Normal University, Huainan, Anhui, 232038, China.
Biochem Biophys Rep. 2021 May 25;26:101026. doi: 10.1016/j.bbrep.2021.101026. eCollection 2021 Jul.
Cancer cell spheroids have been shown to be more physiologically relevant to native tumor tissue than monolayer 2D culture cells. Due to enhanced intercellular communications among cells in spheroids, spheroid secreted exosomes which account for transcellular transportation should exceed those from 2D cell culture and may display a different expression pattern of miRNA or protein. To test this, we employed a widely used pancreatic cancer cell line, PANC-1, to create 3D spheroids and compared exosomes generated by both 2D cell culture and 3D PANC-1 spheroids. We further measured and compared exosomal miRNA and GPC-1 protein expression with qRT-PCR and enzyme-linked immunosorbent assay, respectively. It showed that PANC-1 cells cultured in 3D spheroids can produce significantly more exosomes than PANC-1 2D cells and exosomal miRNA and GPC-1 expression derived from spheroids show more features relevant to the progression of pancreatic cancer. These findings point to the potential importance of using spheroids as model to study cancer development and progression.
与单层二维培养细胞相比,癌细胞球体已被证明在生理上与天然肿瘤组织更相关。由于球体中细胞间通讯增强,负责跨细胞运输的球体分泌外泌体应超过二维细胞培养产生的外泌体,并且可能表现出不同的miRNA或蛋白质表达模式。为了验证这一点,我们使用了一种广泛使用的胰腺癌细胞系PANC-1来创建三维球体,并比较二维细胞培养和三维PANC-1球体产生的外泌体。我们进一步分别通过qRT-PCR和酶联免疫吸附测定法测量和比较外泌体miRNA和GPC-1蛋白表达。结果表明,三维球体培养的PANC-1细胞比二维PANC-1细胞能产生显著更多的外泌体,并且球体来源的外泌体miRNA和GPC-1表达显示出更多与胰腺癌进展相关的特征。这些发现表明使用球体作为研究癌症发展和进展模型的潜在重要性。