PKI-587 and sorafenib alone and in combination on inhibition of liver cancer stem cell proliferation

Roberto Gedaly, Roberto Galuppo, Yolanda Musgrave, Paul Angulo, Jonathan Hundley, Malay Shah, Michael F. Daily, Changguo Chen, Donald A. Cohen, Brett T. Spear, B. Mark Evers

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Background: Deregulated Ras/Raf/mitogen-activated protein kinase and PI3 K/AKT/mTOR signaling pathways are significant in hepatocellular carcinoma proliferation (HCC). In this study we evaluated differences in the antiproliferative effect of dual PI3 K/Akt/mTOR and Ras/Raf/mitogen-activated protein kinase inhibition of non liver cancer stem cell lines (PLC and HuH7) and liver cancer stem cell (LCSC) lines (CD133, CD44, CD24, and aldehyde dehydrogenase 1-positive cells). Materials and methods: Flow cytometry was performed on the resulting tumors to identify the LCSC markers CD133, CD44, CD24, and aldehyde dehydrogenase 1. Methylthiazol tetrazolium assay was used to assess cellular proliferation. Finally, a Western blot assay was used to evaluate for inhibition of specific enzymes in these two signaling pathways. Results: Using flow cytometry, we found that LCSC contain 64.4% CD133 + cells, 83.2% CD44 + cells, and 96.4% CD24 + cells. PKI-587 and sorafenib caused inhibiton of LCSC and HCC cell proliferation. PLC cells were more sensitive to PKI-587 than LCSC or Huh7 (P < 0.001). Interestingly, HuH7 cells were more sensitive to sorafenib than LCSC or PLC cells. Additionally, combination therapy with PKI-587 and sorafenib caused significantly more inhibition than monotherapy in HuH7, PLC, and LCSC. Using the methylthiazol tetrazolium assay, we found that the LCSC proliferation was inhibited with sorafenib monotherapy 39% at 5 μM (P < 0.001; n = 12) and 67% by PKI-587 at 0.1 μM (P = 0.002, n = 12) compared with control. The combination of PKI-587 and sorafenib, however, synergistically inhibited LCSC proliferation by 86% (P = 0.002; n = 12). Conclusions: LCSC (CD133+, CD44+, CD24+) were able to develop very aggressive tumors with low cell concentrations at 4 to 6 wk. Cells CD133+, CD44+, CD24+, which demonstrated at least moderate resistance to therapy in vitro. The combination of PKI-587 and sorafenib was better than either drug alone at inhibiting of LCSC and on HCC cell proliferation.

Original languageEnglish
Pages (from-to)225-230
Number of pages6
JournalJournal of Surgical Research
Volume185
Issue number1
DOIs
StatePublished - Nov 2013

Bibliographical note

Funding Information:
The project described was supported by the National Center for Research Resources and the National Center for Advancing Translational Sciences , National Institutes of Health , through grant UL1TR000117 . The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Keywords

  • Hepatocellular carcinoma
  • Liver cancer stem cells
  • PKI-587
  • Sorafenib

ASJC Scopus subject areas

  • Surgery

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