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Open Access Highly Accessed Research article

3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing

Paola Longati12, Xiaohui Jia12, Johannes Eimer12, Annika Wagman12, Michael-Robin Witt3, Stefan Rehnmark3, Caroline Verbeke4, Rune Toftgård2, Matthias Löhr12* and Rainer L Heuchel12

Author Affiliations

1 CLINTEC, Karolinska Institutet, Stockholm 14186, Sweden

2 Center of Biosciences, Karolinska Institutet, Stockholm 14186, Sweden

3 Axcentua Pharmaceuticals AB, Nobels Allé 10, Stockholm, 14157, Sweden

4 Department of Pathology, Karolinska Institutet, Stockholm, 14186, Sweden

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BMC Cancer 2013, 13:95  doi:10.1186/1471-2407-13-95

Published: 27 February 2013

Abstract

Background

Pancreatic ductal adenocarcinoma (PDAC) is the fourth most common cause of cancer related death. It is lethal in nearly all patients, due to an almost complete chemoresistance. Most if not all drugs that pass preclinical tests successfully, fail miserably in the patient. This raises the question whether traditional 2D cell culture is the correct tool for drug screening. The objective of this study is to develop a simple, high-throughput 3D model of human PDAC cell lines, and to explore mechanisms underlying the transition from 2D to 3D that might be responsible for chemoresistance.

Methods

Several established human PDAC and a KPC mouse cell lines were tested, whereby Panc-1 was studied in more detail. 3D spheroid formation was facilitated with methylcellulose. Spheroids were studied morphologically, electron microscopically and by qRT-PCR for selected matrix genes, related factors and miRNA. Metabolic studies were performed, and a panel of novel drugs was tested against gemcitabine.

Results

Comparing 3D to 2D cell culture, matrix proteins were significantly increased as were lumican, SNED1, DARP32, and miR-146a. Cell metabolism in 3D was shifted towards glycolysis. All drugs tested were less effective in 3D, except for allicin, MT100 and AX, which demonstrated effect.

Conclusions

We developed a high-throughput 3D cell culture drug screening system for pancreatic cancer, which displays a strongly increased chemoresistance. Features associated to the 3D cell model are increased expression of matrix proteins and miRNA as well as stromal markers such as PPP1R1B and SNED1. This is supporting the concept of cell adhesion mediated drug resistance.