PKCε inhibits isolation and stemness of side population cells via the suppression of ABCB1 transporter and PI3K/Akt, MAPK/ERK signaling in renal cell carcinoma cell line 769P

Cancer Lett. 2016 Jun 28;376(1):148-54. doi: 10.1016/j.canlet.2016.03.041. Epub 2016 Mar 29.

Abstract

Protein kinase C epsilon (PKCε), a member of the novel PKC family, is known to be a transforming oncogene and tumor biomarker for many human solid cancers including renal cell carcinoma (RCC). We isolated side population (SP) cells from the RCC 769P cell line, and proved that those cells possess cancer stem cell (CSC) characteristics. In this study, to identify the function of PKCε in cancer stemness of 769P SP cells, we reduced the expression of PKCε in those cells, following the results demonstrated that PKCε depletion had a negative correlation with the existence of SP cells in 769P cell line. Down-regulation of PKCε also suppresses the CSC potential of sorted 769P SP cells in several ways: proliferation potential, resistance to chemotherapeutics and in vivo tumor formation ability. Our study also reveals that PKCε is associated with ABCB1 and this association probably contributed to the SP cells isolation from 769P cell line. Furthermore, the expression of ABCB1 is directly regulated by PKCε. Additionally, after the depletion of PKCε, the phosphorylation of pAkt, pStat3 and pERK was apparently suppressed in 769P SP cells, whereas PKCε overexpression could promote the phosphorylation of AKT, STAT3 and ERK in 769P Non-SP cells. Overall, PKCε down-regulation suppresses sorting and the cancer stem-like phenotype of RCC 769P SP cells through the regulation of ABCB1 transporter and the PI3K/Akt, Stat3 and MAPK/ERK pathways that are dependent on the phosphorylation effects. Thus, PKCε may work as an important mediator in cancer stem cell pathogenesis of renal cell cancer.

Keywords: ABCB1; Cancer stem cell; Protein kinase C epsilon; Renal cell carcinoma; Side population cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / metabolism
  • Animals
  • Carcinoma, Renal Cell / drug therapy
  • Carcinoma, Renal Cell / enzymology*
  • Carcinoma, Renal Cell / pathology
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Separation / methods*
  • Dose-Response Relationship, Drug
  • Down-Regulation
  • Drug Resistance, Neoplasm
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Neoplastic
  • Heterografts
  • Humans
  • Kidney Neoplasms / drug therapy
  • Kidney Neoplasms / enzymology*
  • Kidney Neoplasms / pathology
  • Mice, Inbred NOD
  • Mice, SCID
  • Neoplasm Transplantation
  • Neoplastic Stem Cells / drug effects
  • Neoplastic Stem Cells / enzymology*
  • Neoplastic Stem Cells / pathology
  • Phenotype
  • Phosphatidylinositol 3-Kinase / metabolism*
  • Phosphorylation
  • Protein Kinase C-epsilon / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA Interference
  • STAT3 Transcription Factor / metabolism
  • Side-Population Cells / drug effects
  • Side-Population Cells / enzymology*
  • Side-Population Cells / pathology
  • Signal Transduction
  • Time Factors
  • Transfection

Substances

  • ABCB1 protein, human
  • ATP Binding Cassette Transporter, Subfamily B
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Phosphatidylinositol 3-Kinase
  • Proto-Oncogene Proteins c-akt
  • PRKCE protein, human
  • Protein Kinase C-epsilon
  • Extracellular Signal-Regulated MAP Kinases