A rapid quantification of invasive phenotype in head and neck squamous cell carcinoma: A novel 3D pillar array system

Sung Yong Choi; Dong Woo Lee; Bokhyun Song; Soo Yoon Kim; Hye Jin Kim; Da-Yong Shin; Bosung Ku; Man Ki Chung

doi:10.1016/j.oraloncology.2020.104807

A rapid quantification of invasive phenotype in head and neck squamous cell carcinoma: A novel 3D pillar array system

Oral Oncol. 2020 Sep:108:104807. doi: 10.1016/j.oraloncology.2020.104807. Epub 2020 May 22.

Authors

Sung Yong Choi¹, Dong Woo Lee², Bokhyun Song¹, Soo Yoon Kim¹, Hye Jin Kim¹, Da-Yong Shin¹, Bosung Ku³, Man Ki Chung⁴

Affiliations

¹ Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.
² Department of Biomedical Engineering, Konyang University, Daejeon, Republic of Korea.
³ Central R & D Center, Medical & Bio Decision (MBD) Co., Ltd, Suwon, Republic of Korea.
⁴ Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea. Electronic address: chungmk@skku.edu.

PMID: 32450501
DOI: 10.1016/j.oraloncology.2020.104807

Abstract

Background: The widely used in vitro invasion assays for head and neck squamous cell carcinoma (HNSCC) are wound healing, transwell, and organotypic assays. However, these are still lab-intensive and time-consuming tasks. For the rapid detection and high throughput screening of invasiveness in 3D condition, we propose a novel spheroid invasion assay using commercially available pillar platform system.

Materials and methods: Using the pillar-based spheroid invasion assay, migration and invasion was evaluated in three patient-derived cells (PDCs) of HNSCC. Immunofluorescence of live cells was used for the quantitative measurement of migratory and invaded cells attached to the pillar. Expression of epithelial-mesenchymal transition (EMT)-related gene (snai1/2) was measured by qRT-PCR. We also tested the impact of drug treatments (cisplatin, docetaxel) on the changes in the invasive phenotype.

Results: All PDCs successfully formed spheroid at 4 days and can be measured invasiveness within 7 days. Intriguingly, one PDC (#1) obtained from the advanced stage showed robust migration, invasion and higher transcription of snai1/2, compared with the other two PDCs. Furthermore, the invasion ratio of the control spheroids was about 70% while the invasion ratios of drug-treated spheroids were lower than 50%, and the difference showed statistical significance (p < 0.01).

Conclusion: The presented spheroid invasion assay using pillar array could be useful for the evaluation of cancer cell behavior and physiology in response to diverse therapeutic drugs.

Keywords: 3D culture; Head and Neck Squamous Cell Carcinoma; Invasion; Spheroids.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Humans
Imaging, Three-Dimensional / methods*
Phenotype
Squamous Cell Carcinoma of Head and Neck / diagnostic imaging*