Expression of Wnt and Notch Signaling Pathways are Increased in Cancer Stem-Like Cells in Metastatic Melanoma Cell Line (A375)

Document Type : Original Article

Authors

1 Master of Science in Animal Biology, Towards Cellular-Tactile Science.Department of Animal Biology, Tow-Transe Science, Science & Culture University, Tehran, Iran.

2 Student MSc in Animal Biology. Towards Cellular-Tangential Sciences.Department of Animal Biology, Tow-Transe Science, Science & Culture University, Tehran, Iran.

3 Group of Stem Cells and Developmental Biology.Department of Stem Cells and Developmental Biology, Cellular Research Center, Stem Cell and Tactical Biology Department, Institute of Biology and Technology of Stem Cells, University of Tehran, Tehran, Iran.

4 Senior Cell and Molecular Expert.Department of Stem Cells and Developmental Biology, Cellular Research Center, Stem Cell and Tactical Biology Department, Institute of Biology and Technology of Stem Cells, University of Tehran, Tehran, Iran.

5 Associate Professor Royan Research Group.Department of Stem Cells and Developmental Biology, Cellular Research Center, Stem Cell and Tactical Biology Department, Institute of Biology and Technology of Stem Cells, University of Tehran, Tehran, Iran.

Abstract

Backgound and Objective: Cancer stem cells assume to be responsible for increasing cancer properties such as migration, and drug sensitivity. A lot of studies revealed signaling pathways are enhanced in cancer stem cells.  Melanoma is well-known as heterogeneous cancer and its severity in treatment. These can be attributed to the existence of cancer stem cells. In this study, we first aimed to separate cancer stem cells (cancer stem-like cells) by sphere formation, characterized them, and second examine the expression of Wnt and Notch signaling pathway genes related to adherent cells.
Subjects and Methods: Adherent cell were cultured in the non-adherent condition with serum-free media; spheres obtained, then sphere formation,  clonogenic assay, expression of CD133 and Nestin proteins, Nanog, NESTIN and Oct4 genes as stem related genes were assessed in comparison to adherent cells. In addition, Notch and Wnt signaling pathways genes in both adherent and spheres cells evaluated.
Results: Sphere formation, clonogenic capacity, expression of Nestin protein, but not CD133 were increased in sphere cells in comparession to adherent cells. They also overexpressed β-catenin, cyclinD1, and c-Myc as Wnt down-stream genes, Notch1, HES1 as Notch down-stream genes, Nanog, and NESTIN as stem-related genes.
Conclusion: These results suggest that sphere culture model could be a proper experimental method to separate cancer stem-like cells. Our data also support two important pathways are overactivated in melanoma cancer stem-like cells which must be considered in targeting therapy.

Keywords

References

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Jundishapur Scientific Medical Journal
Volume 16, Issue 6 - Serial Number 111
January and February 2018
Pages 643-652

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History

  • Receive Date: 09 July 2017
  • Revise Date: 22 November 2017
  • Accept Date: 29 November 2017

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