Treatment of cancer stem cells from human colon adenocarcinoma cell line HT-29 with resveratrol and sulindac induced mesenchymal-endothelial transition rate

  • Ayda Pouyafar
  • Aysa Rezabakhsh
  • Reza RahbarghaziEmail author
  • Milad Zadi Heydarabad
  • Elhameh Shokrollahi
  • Emel Sokullu
  • Majid Khaksar
  • Alireza NourazarianEmail author
  • Çığır Biray AvciEmail author
Regular Article


In the current experiment, the combined regime of resveratrol and a Wnt-3a inhibitor, sulindac, were examined on the angiogenic potential of cancer stem cells from human colon adenocarcinoma cell line HT-29 during 7 days. Cancer stem cells were enriched via a magnetic-activated cell sorter technique and cultured in endothelial induction medium containing sulindac and resveratrol. Expression of endothelial markers such as the von Willebrand factor (vWF) and vascular endothelial cadherin (VE-cadherin) and genes participating in mesenchymal-to-epithelial transition was studied by real-time PCR assay. Protein levels of Wnt-3a and angiogenic factor YKL-40 were examined by western blotting. ELISA was used to determine the level of N-acetylgalactosaminyltransferase 11 (GALNT11) during mesenchymal-endothelial transition. Autophagy status was monitored by PCR array under treatment with the resveratrol plus sulindac. Results showed that resveratrol and sulindac had the potential to decrease the cell survival of HT-29 cancer cells and the clonogenic capacity of cancer stem cells compared with the control (p < 0.05). The expression of VE-cadherin and vWF was induced in cancer stem cells incubated with endothelial differentiation medium enriched with resveratrol (p < 0.05). Interestingly, the Wnt-3a level was increased in the presence of resveratrol and sulindac (p < 0.05). YKL-40 was reduced after cell exposure to sulindac and resveratrol. The intracellular content of resistance factor GALNT11 was diminished after treatment with resveratrol (p < 0.05). Resveratrol had the potential to induce the transcription of autophagy signaling genes in cancer stem cells during endothelial differentiation (p < 0.05). These data show that resveratrol could increase cancer stem cell trans-differentiation toward endothelial lineage while decrease cell resistance by modulation of autophagy signaling and GALNT11 synthesis.


Human cancer stem cells Sulindac Resveratrol Endothelial differentiation Autophagy 



The authors wish to thank the personnel of the Stem Cell Research Center of Tabriz University of Medical Sciences for their kindest help and guidance.

Author contributions

Ayda Pouyafar: Original draft/data collection, cancer stem cell isolation by MACS, cell culture. Aysa Rezabakhsh: Western blotting. Milad Zadi Heydarabad: Primer design and real-time PCR analysis. Emel Sokullu: Manuscript preparation and edition. Majid Khaksar: ELISA. Alireza Nourazarian: Equal conceptualization and cell culture. Çığır Biray Avci: PCR array analysis and interpretation. Reza Rahbarghazi: Equal conceptualization and data correction and supervision of all procedure.


This study was supported by a grant from the Tabriz University of Medical Sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Stem Cell Research CenterTabriz University of Medical SciencesTabrizIran
  2. 2.Aging Research InstituteTabriz University of Medical SciencesTabrizIran
  3. 3.Department of Applied Cell Sciences, Faculty of Advanced Medical SciencesTabriz University of Medical SciencesTabrizIran
  4. 4.Medicinal Plants Research CenterYasuj University of Medical SciencesYasujIran
  5. 5.Bioengineering DepartmentIzmir Katip Celebi UniversityIzmirTurkey
  6. 6.Harvard Medical School, Division of Biomedical Engineering at Brigham and Women’s HospitalHarvard-MIT Health Sciences and TechnologyCambridgeUSA
  7. 7.Department of Biochemistry and Clinical Laboratories, Faculty of MedicineTabriz University of Medical SciencesTabrizIran
  8. 8.Faculty of Medicine, Department of Medical BiologyEge UniversityIzmirTurkey

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