Scrophularia Atropatana Extract Reverses TP53 Gene Promoter Hypermethylation and Decreases Survivin Antiapoptotic Gene Expression in Breast Cancer Cells

Ghavifekr Fakhr, Mehrdad; Rezaie Kahkhaie, Kolsoum; Shanehbandi, Dariush; Farshdousti Hagh, Majid; Zarredar, Habib; Safarzadeh, Elham; Abdolrahimi Vind, Mina; Baradaran, Behzad

doi:10.22034/APJCP.2018.19.9.2599

Scrophularia Atropatana Extract Reverses TP53 Gene Promoter Hypermethylation and Decreases Survivin Antiapoptotic Gene Expression in Breast Cancer Cells

Document Type : Research Articles

Authors

¹ Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

² Student Research Committee,Zabol University of Medical Science, Zabol, Iran.

³ Department of Biochemistry, Faculty of Medical Sciences, Zabul University, Zabol, Iran.

⁴ Zabol Medicininal Plants Research Center, Zabol University of Medical sciences, Zabol, Iran.

10.22034/APJCP.2018.19.9.2599

Abstract

Background: In many cases of breast cancer, the aberrant methylation of TP53 gene leads to uncontrolled cell
proliferation and apoptosis inhibition. Moreover, expression of oncogenes which are under the control of P53 protein
could be altered. Survivin as a conspicuous example of this category plays important roles in tumorigenesis, drug
resistance and apoptosis inhibition. The present study was done to reveal the effects of Scrophularia atropatana extract
on epigenetic situation of TP53 gene promoter and the expression levels of anti-apoptotic gene, survivin and its potential
for production of cancer epi-drugs. Methods: Cytotoxic effect of dichloromethane extracts of Scrophularia plant on
MCF-7 cell line was assessed in our previous study. Cell death ELISA (enzyme-linked immunosorbent assay) and
TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) tests were used to investigate the occurrence of
apoptosis in the treated cells. Methylation Specific PCR (MSP) was employed to assess the changes in methylation
status of the TP53 gene promoter. Furthermore, quantitative real time PCR was utilized to evaluate the resulting changes
in TP53 and survivin genes expression. Results: Cell death ELISA and TUNEL assays confirmed the occurrence of
apoptosis. MSP test revealed a significant change in the methylation status of TP53 promoter. QRT-PCR showed
an increased TP53 gene expression in the treated cells while a significant decrease in survivin mRNA was evident.
Conclusions: According to the outcomes, dichloromethane extract of S. atropatana returned the TP53 gene promoter
hypermethylation to normal state. This plant could be a promising source for production of epi-drugs due to its apoptotic
effects and reversal of TP53 epigenetic alterations.

Keywords