TITLE:
The Evaluation of the Effects of Temozolomide on MGMT Gene Expression in MCF-7 and SKBR3 Human Breast Cancer Cell Lines
AUTHORS:
Onur Eroğlu, Büşra Sevim
KEYWORDS:
Breast Cancer, Temozolomide, MGMT, MCF-7, SKBR3, HRM
JOURNAL NAME:
Journal of Cancer Therapy,
Vol.10 No.3,
March
11,
2019
ABSTRACT: Background
and Aim: In this study, it was aimed
to examine the cytotoxic effect of temozolomide (TMZ) treatment, on MCF-7 and
SKBR3 cell lines, to study the methylation levels of MGMT gene expression and
gene promoter region. Methods: The MTT test was performed to determine the
effective dose of TMZ. The time-dependent cell survival test was performed
after the IC50 value was found. Western blotting was performed to
determine MGMT gene expression levels. High Resolution Melting (HRM) technique
was used to determine the methylation levels of MGMT gene promoter region. Results: TMZ has been shown to have a high cytotoxic effect on SKBR3 cell line and low
cytotoxicity on MCF-7. When MGMT expression levels before and after TMZ
treatment were observed by western blotting, the gene expression levels of TMZ
treatment were shown to decrease in both cell lines. It was observed that MGMT
gene promoter region was hypermethylated in two cell lines, and that the application of TMZ further increased the methylation levels in the promoter region. Conclusions: It was seen that TMZ could be used as a single agent in SKBR-3 cell line. With
this study on breast cancer, it is expected that temozolomide treatment will
lead future in vitro and in vivo studies for breast cancer.