Establishment of a Novel Model for Anticancer Drug Resistance in Three-Dimensional Primary Culture of Tumor Microenvironment.

Tatsuya Usui,Nobuyuki Fujiwara,Masao Nakajima,Hiroaki Nagano,Nobuaki Suzuki,Ryotaro Yabe,Shuhei Enjoji,Takashi Ohama,Shunya Tsuji,Masashi Sakurai,Shoichi Hazama,Hideyoshi Kawasaki,Koji Umata,Ryouichi Tsunedomi,Hiroko Takenouchi,Hideyuki Yamawaki,Koichi Sato

doi:10.1155/2016/7053872

Tatsuya Usui, Nobuyuki Fujiwara + Show 15 more

Open Access

https://doi.org/10.1155/2016/7053872

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Abstract

Tumor microenvironment has been implicated in tumor development and progression. As a three-dimensional tumor microenvironment model, air liquid interface (ALI) organoid culture from oncogene transgenic mouse gastrointestinal tissues was recently produced. However, ALI organoid culture system from tissues of colorectal cancer patients has not been established. Here, we developed an ALI organoid model from normal and tumor colorectal tissues of human patients. Both organoids were successfully generated and showed cystic structures containing an epithelial layer and surrounding mesenchymal stromal cells. Structures of tumor organoids closely resembled primary tumor epithelium. Expression of an epithelial cell marker, E-cadherin, a goblet cell marker, MUC2, and a fibroblast marker, vimentin, but not a myofibroblast marker, α-smooth muscle actin (SMA), was observed in normal organoids. Expression of E-cadherin, MUC2, vimentin, and α-SMA was observed in tumor organoids. Expression of a cancer stem cell marker, LGR5 in tumor organoids, was higher than that in primary tumor tissues. Tumor organoids were more resistant to toxicity of 5-fluorouracil and Irinotecan than colorectal cancer cell lines, SW480, SW620, and HCT116. These findings indicate that ALI organoid culture from colorectal cancer patients may become a novel model that is useful for examining resistance to chemotherapy in tumor microenvironment.

Highlights

Colorectal cancer is one of the most common cancers and is one of the leading causes of morbidity and mortality all over the world [1]
We observed that normal air liquid interface (ALI) organoids consisted of monolayer of epitheliallike cells and the surrounding stromal-like cells in each patient culture (Figures 1(c) and 1(d))
We observed that the number of tumor ALI organoid colonies significantly increased compared with it just after passaging (Figure 2(b)) and the different structures of tumor organoids in each patient culture (Figure 2(c))

Summary

Introduction

Colorectal cancer is one of the most common cancers and is one of the leading causes of morbidity and mortality all over the world [1]. Primary tumor tissues contain a few cancer stem cells (CSCs), which have the capacity to metastasize and cause resistance to chemotherapy [3]. Most of the in vitro studies in colorectal cancer research have been performed using a two-dimensional (2D) cell culture model. It hardly reflects cellular heterogeneity and behavior of tissues in vivo. High throughput screening in drug discovery was performed using tumor organoids. These reports suggest a possibility that the technology of Matrigel organoid culture is applied to a personalized therapy for colorectal cancer in near future

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