Culturing conditions highly affect DNA methylation and gene expression levels in MCF7 breast cancer cell line

  • Lama Hamadneh
  • May Al-Majawleh
  • Yazun Jarrar
  • Sawsan Shraim
  • Mariam Hasan
  • Basha’er Abu-Irmaileh


The levels of DNA methylation and their role in gene expression are key factors that could affect diagnosis, prognosis, and treatment options of different diseases. In this study, the methylation levels of 22 genes that are mostly correlated to breast cancer were determined using EpiTect methyl II PCR array. This analysis was performed to determine the effect of cells’ passage number and the use of antibiotics in the culturing media on gene methylation levels in MCF7 cell line. DNA methylation levels of PTGS2, ADAM23, HIC1, and PYCARD were found to be significantly different among different passages. While the DNA methylation levels of CCNA1, RASSF1, and THBS1 were found to be affected by the use of 1% of penicillin/streptomycin in the culture media. Gene expression analysis after demethylation using 5-Aza-2′-deoxycytidine showed that the gene expression levels of the hypermethylated genes varied between different passage numbers. This study shows that the presence of antibiotic within cultured media and cell line’s passage number could greatly affect the methylation levels that need to be considered in future studies on cell lines.


DNA methylation Breast cancer mcf-7 cell line Passage number Penicillin/streptomycin 


Funding information

This study received a financial support from Al-Zaytoonah University of Jordan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


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Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  1. 1.Faculty of PharmacyAl-Zaytoonah University of JordanAmmanJordan
  2. 2.Hamdi Mango Center for Scientific ResearchUniversity of JordanAmmanJordan

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