A cancer invasion model of cancer-associated fibroblasts aggregates combined with TGF-β1 release system

Teruki Nii; Kimiko Makino; Yasuhiko Tabata

doi:10.1016/j.reth.2020.02.003

A cancer invasion model of cancer-associated fibroblasts aggregates combined with TGF-β1 release system

Regen Ther. 2020 Mar 4:14:196-204. doi: 10.1016/j.reth.2020.02.003. eCollection 2020 Jun.

Authors

Teruki Nii^{1

2}, Kimiko Makino^{2

3}, Yasuhiko Tabata¹

Affiliations

¹ Laboratory of Biomaterials, Institute for Frontier Life and Medical Sciences, Kyoto University, 53 Kawara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.
² Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641, Yamazaki, Noda, 278-8510, Japan.
³ Center for Drug Delivery Research, Tokyo University of Science, 2641, Yamazaki, Noda, 278-8510, Japan.

Abstract

Introduction: The objective of this study is to design a cancer invasion model where the cancer invasion rate can be regulated in vitro.

Methods: Cancer-associated fibroblasts (CAF) aggregates incorporating gelatin hydrogel microspheres (GM) containing various concentrations of transforming growth factor-β1 (TGF-β1) (CAF-GM-TGF-β1) were prepared. Alpha-smooth muscle actin (α-SMA) for the CAF aggregates was measured to investigate the CAF activation level by changing the concentration of TGF-β1. An invasion assay was performed to evaluate the cancer invasion rate by co-cultured of cancer cells with various CAF-GM-TGF-β1.

Results: The expression level of α-SMA for CAF increased with an increased in the TGF-β1 concentration. When co-cultured with various types of CAF-GM-TGF-β1, the cancer invasion rate was well correlated with the α-SMA level. It is conceivable that the TGF-β1 concentration could modify the level of CAF activation, leading to the invasion rate of cancer cells. In addition, at the high concentrations of TGF-β1, the effect of a matrix metalloproteinase (MMP) inhibitor on the cancer invasion rate was observed. The higher invasion rate would be achieved through the higher MMP production.

Conclusions: The present model is promising to realize the cancer invasion whose rate can be modified by changing the TGF-β1 concentration.

Keywords: 2D, two-dimensional; 3D, three-dimensional; Anti-cancer drug screening; CAF, cancer-associated fibroblasts; Cancer invasion model; DDW, double-distilled water; Drug delivery system; ELISA, enzyme-linked immunosolvent assay; FCS, fetal calf serum; GM, gelatin hydrogel microspheres; Gelatin hydrogel microspheres; MEM, minimum essential medium; MMP, matrix metalloproteinase; PBS, phosphate buffered-saline; PLGA, poly (lactic-co-glycolic acid); PVA, poly (vinyl alcohol); TGF-β1, transforming growth factor-β1; Three-dimensional cell culture; α-SMA, alpha-smooth muscle actin.