Influence of adriamycin on changes in Nanog, Oct-4, Sox2, ARID1 and Wnt5b expression in liver cancer stem cells

World J Gastroenterol. 2014 Jun 14;20(22):6974-80. doi: 10.3748/wjg.v20.i22.6974.

Abstract

Aim: To determine the influence of Adriamycin (ADM) on the changes in Nanog, Oct4, Sox2, as well as, in ARID1 and Wnt5b expression in liver cancer stem cells.

Methods: The MHCC97-L and HCCLM3 liver cancer cell lines were selected as the cell models in this study, and were routinely cultured. The 50% lethal dose (LD50) in the cell lines was detected by the MTT assay. Expression changes in liver cancer stem cell related genes (Nanog, Oct-4, Sox2, ARID1, and Wnt5b) were detected by western blot following treatment with ADM (LD50).

Results: The LD50 of ADM in MHCC97-L cells was lower than that in HCCLM3 cells (0.4123 ± 0.0236 μmol/L vs 0.5259 ± 0.0125 μmol/L, P < 0.05). Wnt5b and Nanog were expressed in both MHCC97-L and HCCLM3 cells, while only Sox2 was expressed in HCCLM3 cells. However, neither ARID1A nor Oct4 was detected in these two cell lines. Genes, related to the stem cells, showed different expression in liver cancer cells with different metastatic potential following treatment with ADM (LD50). Wnt5b protein increased gradually within 4 h of ADM (LD50) treatment, while Nanog decreased (P < 0.05). After 12 h, Wnt5b decreased gradually, while Nanog increased steadily (P < 0.05). In addition, only Sox2 was expressed in HCCLM3 cells with high metastatic potential following ADM (LD50) treatment. The expression of Sox2 increased gradually with ADM (LD50) in HCCLM3 cells (P < 0.05).

Conclusion: ADM increased the death rate of MHCC97-L and HCCLM3 cells, while the growth suppressive effect of ADM was higher in MHCC97-L cells than in HCCLM3 cells.

Keywords: Adriamycin; Liver cancer cell; Liver cancer stem cell; Stem cell related gene; Western blot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibiotics, Antineoplastic / pharmacology*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • DNA-Binding Proteins
  • Dose-Response Relationship, Drug
  • Doxorubicin / pharmacology*
  • Homeodomain Proteins / metabolism*
  • Humans
  • Inhibitory Concentration 50
  • Liver Neoplasms / metabolism*
  • Liver Neoplasms / pathology
  • Nanog Homeobox Protein
  • Neoplastic Stem Cells / drug effects*
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / pathology
  • Nuclear Proteins / metabolism*
  • Octamer Transcription Factor-3 / metabolism*
  • SOXB1 Transcription Factors / metabolism*
  • Time Factors
  • Transcription Factors / metabolism*
  • Wnt Proteins / metabolism*

Substances

  • ARID1A protein, human
  • Antibiotics, Antineoplastic
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Nuclear Proteins
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Transcription Factors
  • WNT5B protein, human
  • Wnt Proteins
  • Doxorubicin