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Gastric Cancer

, Volume 22, Issue 5, pp 967–979 | Cite as

Identification of different gene expressions between diffuse- and intestinal-type spheroid-forming gastric cancer cells

  • Jong Won Lee
  • Jae Sook Sung
  • Young Soo Park
  • Seok Chung
  • Yeul Hong KimEmail author
Original Article
  • 362 Downloads

Abstract

Background

Three-dimensional in vitro spheroid models are unique because they are considered for enrichment of specific cell populations with self-renewal ability. In this study, we explored the different mechanisms of gastric cancer spheroid-forming cells according to the Lauren classification.

Methods

We isolated and enriched cells with self-renewal ability using spheroid-forming methods from gastric cancer cell lines. The expression of candidate target genes was investigated using western blot and qRT-PCR analysis. Lentiviral shRNA knockdown of target gene expression was performed and the effects on spheroid, colony forming, and tumorigenic ability were analyzed.

Results

The SNU-638, SNU-484, MKN-28, and NCI-N87 successfully formed spheroid from single cell and enriched for self-renewal ability from 11 gastric cancer cell lines, including diffuse and intestinal types. The expression of SOX2 and E-cadherin increased in spheroid-forming cells in a diffuse-type cell line (SNU-638 and SNU-484), but not in the intestinal type (MKN-28 and NCI-N87). In contrast, ERBB3 expression was only increased in intestinal-type spheroid cells. The depletion of each candidate target gene expression suppressed self-renewal ability to grow as spheroids and colonies in a soft agar assay. In particular, down-regulated ERBB3 in the intestinal-type cell lines inhibited tumor growth in a mouse xenograft model. We found that high ERBB3 gene expression correlates with decreased survival in the intestinal type of gastric cancer.

Conclusions

Our results suggest that diffuse- and intestinal-type spheroid-forming cells express genes differently. Our data suggest that these candidate genes from spheroid-forming cells can be used in applications in targeted therapy.

Keywords

Gastric cancer Cellular spheroid Lauren classification Diffuse type Intestinal type 

Notes

Acknowledgements

This study was supported by a Grant from the National Research Foundation of Korea (NRF), which is funded by the Korean government (MEST) (No. 2010-0020986).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Animal studies

All institutional and national guidelines for the care and use of laboratory animals were followed.

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Copyright information

© The International Gastric Cancer Association and The Japanese Gastric Cancer Association 2019

Authors and Affiliations

  • Jong Won Lee
    • 1
    • 3
  • Jae Sook Sung
    • 2
  • Young Soo Park
    • 2
  • Seok Chung
    • 4
  • Yeul Hong Kim
    • 1
    • 2
    • 3
    Email author
  1. 1.Brain Korea 21 Plus Project for Biomedical ScienceKorea University College of MedicineSeoulRepublic of Korea
  2. 2.Cancer Research InstituteKorea University College of MedicineSeoulRepublic of Korea
  3. 3.Division of Oncology/Hematology, Department of MedicineKorea University Anam Hospital, Korea University College of MedicineSeoulRepublic of Korea
  4. 4.School of Mechanical EngineeringKorea UniversitySeoulRepublic of Korea

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