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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.

Titel:	Combined evaluation of proliferation and apoptosis to calculate IC <subscript>50</subscript> of VPA-induced PANC-1 cells and assessing its effect on the Wnt signaling pathway.
Autor/in / Beteiligte Person:	Ekici, Y ; Yilmaz, A ; Kucuksezer, UC ; Gazioglu, SB ; Yamalioglu, ZD ; Gurol, AO ; Linn, T ; Tuncer, FN
Link:	Zum Volltext
Zeitschrift:	Medical oncology (Northwood, London, England), Jg. 38 (2021-08-06), Heft 9, S. 109
Veröffentlichung:	2011- : New York : Springer ; <i>Original Publication</i>: Northwood, Middlesex, England : Science and Technology Letters, c1994-, 2021
Medientyp:	academicJournal
ISSN:	1559-131X (electronic)
DOI:	10.1007/s12032-021-01560-4
Schlagwort:	Anticonvulsants pharmacology Carcinoma, Pancreatic Ductal drug therapy Carcinoma, Pancreatic Ductal genetics Carcinoma, Pancreatic Ductal metabolism Humans Inhibitory Concentration 50 Lymphoid Enhancer-Binding Factor 1 genetics Lymphoid Enhancer-Binding Factor 1 metabolism Pancreatic Neoplasms drug therapy Pancreatic Neoplasms genetics Pancreatic Neoplasms metabolism Tumor Cells, Cultured Apoptosis Carcinoma, Pancreatic Ductal pathology Cell Proliferation Gene Expression Regulation, Neoplastic drug effects Pancreatic Neoplasms pathology Valproic Acid pharmacology Wnt Signaling Pathway drug effects
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Evaluation Study; Journal Article Language: English [Med Oncol] 2021 Aug 06; Vol. 38 (9), pp. 109. <i>Date of Electronic Publication: </i>2021 Aug 06. MeSH Terms: Apoptosis* ; Cell Proliferation* ; Carcinoma, Pancreatic Ductal / pathology ; Gene Expression Regulation, Neoplastic / drug effects ; Pancreatic Neoplasms / pathology ; Valproic Acid / pharmacology ; Wnt Signaling Pathway / drug effects ; Anticonvulsants / pharmacology ; Carcinoma, Pancreatic Ductal / drug therapy ; Carcinoma, Pancreatic Ductal / genetics ; Carcinoma, Pancreatic Ductal / metabolism ; Humans ; Inhibitory Concentration 50 ; Lymphoid Enhancer-Binding Factor 1 / genetics ; Lymphoid Enhancer-Binding Factor 1 / metabolism ; Pancreatic Neoplasms / drug therapy ; Pancreatic Neoplasms / genetics ; Pancreatic Neoplasms / metabolism ; Tumor Cells, Cultured References: McGuire S, World Cancer Report. 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(PMID: 10.1371/journal.pone.0001155179825072048934) Grant Information: 32763 Bilimsel Araştirma Projeleri Birimi, Istanbul Üniversitesi Contributed Indexing: Keywords: Apoptosis; IC50; Pancreatic ductal adenocarcinoma; Proliferation; Valproic acid; Wnt signaling Substance Nomenclature: 0 (Anticonvulsants) ; 0 (LEF1 protein, human) ; 0 (Lymphoid Enhancer-Binding Factor 1) ; 614OI1Z5WI (Valproic Acid) Entry Date(s): Date Created: 20210806 Date Completed: 20220103 Latest Revision: 20220103 Update Code: 20240513

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