PlGF Knockdown Decreases Tumorigenicity and Stemness Properties of Spheroid Body Cells Derived from Gastric Cancer Cells

J Cell Biochem. 2017 Apr;118(4):851-859. doi: 10.1002/jcb.25762. Epub 2016 Oct 25.

Abstract

Placental growth factor (PlGF) a member of the vascular endothelial growth factor family regulates some cell processes such as survival, growth of vascular endothelial cells, invasiveness, and also involves in pathological angiogenesis and metastasis in most cancers. Cancer stem cells are believed to be the main reason for the tumor relapse and resistance to therapy. These cells have various characteristics as same as normal tissue-specific adult stem cells including self-renewability and potent to differentiate into various cell types. However, the function of PlGF in gastric cancer stem cells is not well understood. We have investigated the effect of PlGF knockdown on the tumorigenicity and stem cell properties of spheroid body cells derived from two human gastric cancer cell lines. In this study, we isolated spheroid body cells which have stemness properties from MKN-45 and AGS without using growth factors. Validation of spheroid body cells was confirmed by various methods. Then the effects of PlGF knockdown were investigated on in vitro tumorigenicity, differentiation, migration, angiogenesis, and transcription levels of stemness markers of spheroid body cells. Our findings indicated that isolation of spheroid body cells from MKN-45 and AGS cells without using growth factors is an easy and inexpensive method to isolate cancer stem cells and knockdown of PlGF in spheroid body cells reduced in vitro tumorigenicity and stemness properties of spheroid body cells such as Self-renewal ability, colony forming, migratory, and MMPs activities and decreased ability to differentiation and angiogenesis. J. Cell. Biochem. 118: 851-859, 2017. © 2016 Wiley Periodicals, Inc.

Keywords: CANCER STEM CELLS; GASTRIC CANCER; PLGF; SPHEROID BODY CELLS (SBCS); STEMNESS; TUMORIGENICITY.

MeSH terms

  • Cell Differentiation / genetics
  • Cell Differentiation / physiology
  • Cell Line, Tumor
  • Gene Knockdown Techniques
  • Genetic Therapy
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Hyaluronan Receptors / genetics
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Membrane Proteins / antagonists & inhibitors*
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology
  • Neoplastic Stem Cells / pathology
  • Neovascularization, Physiologic / genetics
  • Neovascularization, Physiologic / physiology
  • Octamer Transcription Factor-3 / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics
  • SOXB1 Transcription Factors / genetics
  • Spheroids, Cellular / metabolism
  • Spheroids, Cellular / pathology*
  • Stomach Neoplasms / genetics
  • Stomach Neoplasms / pathology
  • Stomach Neoplasms / therapy*
  • Tumor Stem Cell Assay

Substances

  • CD44 protein, human
  • Hyaluronan Receptors
  • Membrane Proteins
  • Octamer Transcription Factor-3
  • PIGF protein, human
  • POU5F1 protein, human
  • RNA, Messenger
  • RNA, Small Interfering
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • MMP2 protein, human
  • Matrix Metalloproteinase 2
  • MMP9 protein, human
  • Matrix Metalloproteinase 9