TITLE:
Investigation of Methylation Profiles of TP53, Caspase 9, Caspase 8, Caspase 3 Genes Treated with DNA Methyl Transferase Inhibitor (DNMTi) Zebularine (ZEB) and Caffeic Acid Phenethyl Ester (CAPE) on MCF-7 and MDA-MB-231 Breast Cancer Cell Lines
AUTHORS:
Onur Eroglu, Esin Guvenir Celik, Hacer Kaya, Merve Celen, Mustafa Karabicici, Elif Karacoban
KEYWORDS:
MCF-7, MDA-MB-231, Zebularine, CAPE, Breast Cancer, Methylation
JOURNAL NAME:
Journal of Cancer Therapy,
Vol.10 No.1,
January
23,
2019
ABSTRACT: Purpose: MCF-7 (ER+, WTP53) and
MDA-MB-231 (ER Met, Mutant P53) Caffeic Acid Phenethyl Ester (CAPE) and DNA
Methyl Transferase Inhibitor (DNMTi) in breast cancer cell lines of Zebularine
(ZEB) single and combined application of TP53, caspase-9, caspase 8 and
caspase-3 genes as a result of the use of single and combined drug methylation
profiles are aimed to be evaluated by specific PCR method. Material-Metods: In the MCF-7 and MDA-MB-231 breast cancer cell lines, MTT test and survival
analysis were performed as a result of single and combined
application of CAPE and Zebularine and Methylation
Specific PCR was performed to examine the methylation of caspase-3, caspase-8,
caspase-9 and TP53 genes. Results: According to the results of
24-hour drug administration, the IC50 for the MCF-7 cell line was
determined as 200 μM, for CAPE 40 μM and for the combined values of 50 μM ZEB +
5 μM CAPE. The effects of caspase-3, caspase-8, caspase-9 and TP53 genes on the
methylation level of ZEB, CAPE and ZEB + CAPE drug combination were determined
by using bisulfite modified DNAs in MCF-7 and MDA-MB-231 cell lines. Discussion: In the MCF-7 cell line, the 120 μM ZEB viability rate was 51%, and the
viability of 80 μM ZEB MDA-MB-231 breast cancer cells decreased by 59.7%. After
20 μM CAPE, viability in MCF-7 cells decreased by 31% in 120 μM CAPE and MDA-MB-231 cells decreased by 41%. The viability with 40
μM CAPE decreased by 19% in MDA-MB-231 cells. It was found that 20 μM CAPE
concentration was associated with TP53 methylation in MCF-7 cell lines. The 80
μM ZEB concentration was found to be closely related to the unmethylated status
of the TP53 gene. These results obtained with 50 μM ZEB + 5 μM CAPE application were
found to be related to the methylated-unmetylated status of the TP53 gene in
half (50%). For the caspase-9 gene of MDA-MB-231 cells, 80 μM ZEB concentration
was found to be associated with unmetylated status. The effective use of drugs
with low concentrations of the drug dose provides a more appropriate approach
in terms of treatment.