Combined evaluation of proliferation and apoptosis to calculate IC 50 of VPA-induced PANC-1 cells and assessing its effect on the Wnt signaling pathway.

Autor: Ekici Y; Department of Genetics, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Vakif Gureba Caddesi, Capa, 34093, Istanbul, Turkey.; Graduate School of Health Sciences, Istanbul University, Istanbul, Turkey., Yilmaz A; Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey., Kucuksezer UC; Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey., Gazioglu SB; Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey., Yamalioglu ZD; Cell Processing Unit, Bone Marrow Transplantation Center, Yeni Yuzyil University Gaziosmanpasa Hospital, Istanbul, Turkey.; Department of Medical Microbiology, Medical Faculty, Yeni Yuzyil University, Istanbul, Turkey., Gurol AO; Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey.; Diabetes Application and Research Center, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey., Linn T; Clinical Research Unit, Center of Internal Medicine, Justus-Liebig-University, Giessen, Germany., Tuncer FN; Department of Genetics, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Vakif Gureba Caddesi, Capa, 34093, Istanbul, Turkey. ftuncer@istanbul.edu.tr.; Diabetes Application and Research Center, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey. ftuncer@istanbul.edu.tr.
Jazyk: angličtina
Zdroj: Medical oncology (Northwood, London, England) [Med Oncol] 2021 Aug 06; Vol. 38 (9), pp. 109. Date of Electronic Publication: 2021 Aug 06.
DOI: 10.1007/s12032-021-01560-4
Abstrakt: Pancreatic ductal adenocarcinoma (PDAC) is among the most deadly cancers. Since most patients develop resistance to conventional treatments, new approaches are in urgency. Valproic acid (VPA) was shown to induce apoptosis and reduce proliferation in PANC-1 cells. Wnt signaling pathway is known to be involved in apoptosis and PDAC onset. However, VPA-induced apoptosis and its impact on Wnt signaling in PDACs are not linked, yet. We aimed to calculate IC 50 of VPA-induced PANC-1 cells by combined analyses of proliferation and apoptosis, while assessing its effect on Wnt signaling pathway. PANC-1 was induced with increased VPA doses and time points. Three independent proliferation and apoptosis assays were performed utilizing carboxyfluorescein succinimidyl ester and Annexin V/PI staining, respectively. Flow cytometry measurements were analyzed by CellQuest and NovoExpress. Taqman hydrolysis probes and SYBR Green PCR Mastermix were assessed in expression analyses of Wnt components utilizing 2 -ΔΔCt method. Cell proliferation was inhibited by 50% at 2.5 mM VPA that evoked a significant apoptotic response. Among the screened Wnt components and target genes, only LEF1 exhibited significant four-fold upregulation at this concentration. In conclusion, cancer studies mostly utilize MTT or BrdU assays in estimating cell proliferation and calculating IC 50 of drugs, which provided conflicting VPA dosages utilizing PANC-1 cells. Our novel combined approach enabled specific, accurate and reproducible IC 50 calculation at single cell basis with no apparent effect on Wnt signaling components. Future studies are needed to clarify the role of LEF1 in this model.
(© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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