Three-dimensional culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer

Cunxi Li, Bhuminder Singh, Ramona Graves-Deal, Haiting Ma, Alina Starchenko, William H. Fry, Yuanyuan Lu, Yang Wang, Galina Bogatcheva, Mohseen P. Khan, Ginger L. Milne, Shilin Zhao, Gregory Daniel Ayersk, Nenggan Li, Huaying Hu, Mary Kay Washington, Timothy J. Yeatman, Oliver G. McDonald, Qi Liu, Robert J. Coffey

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

We previously reported that single cells from a human colorectal cancer (CRC) cell line (HCA-7) formed either hollow single-layered polarized cysts or solid spiky masses when plated in 3D in type-I collagen. To begin in-depth analyses into whether clonal cysts and spiky masses possessed divergent properties, individual colonies of each morphology were isolated and expanded. The lines thus derived faithfully retained their parental cystic and spiky morphologies and were termed CC (cystic) and SC (spiky), respectively. Although both CC and SC expressed EGF receptor (EGFR), the EGFRneutralizing monoclonal antibody, cetuximab, strongly inhibited growth of CC, whereas SC was resistant to growth inhibition, and this was coupled to increased tyrosine phosphorylation of MET and RON. Addition of the dual MET/RON tyrosine kinase inhibitor, crizotinib, restored cetuximab sensitivity in SC. To further characterize these two lines, we performed comprehensive genomic and transcriptomic analysis of CC and SC in 3D. One of the most upregulated genes in CC was the tumor suppressor 15-PGDH/HPGD, and the most up-regulated gene in SC was versican (VCAN) in 3D and xenografts. Analysis of a CRC tissue microarray showed that epithelial, but not stromal, VCAN staining strongly correlated with reduced survival, and combined epithelial VCAN and absent HPGD staining portended a poorer prognosis. Thus, with this 3D system, we have identified a mode of cetuximab resistance and a potential prognostic marker in CRC. As such, this represents a potentially powerful system to identify additional therapeutic strategies and disease-relevant genes in CRC and possibly other solid tumors.

Original languageEnglish (US)
Pages (from-to)E2852-2861
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number14
DOIs
StatePublished - Apr 4 2017
Externally publishedYes

Keywords

  • 3d culture
  • Cetuximab resistance
  • Colorecta cancer
  • HPGD
  • Versican

ASJC Scopus subject areas

  • General

Fingerprint

Dive into the research topics of 'Three-dimensional culture system identifies a new mode of cetuximab resistance and disease-relevant genes in colorectal cancer'. Together they form a unique fingerprint.

Cite this