Investıgatıon of the relatıonshıp of DHRS2 gene wıth ubıquıtın genes ın breast cancer

Şakar D. N., Salman Yaylaz B., Punar Ş., Sırma Ekmekci S., Abacı N.

21st International AEK Cancer Congress, Kassel, Almanya, 15 - 17 Şubat 2023, ss.193

  • Yayın Türü: Bildiri / Özet Bildiri
  • Basıldığı Şehir: Kassel
  • Basıldığı Ülke: Almanya
  • Sayfa Sayıları: ss.193
  • İstanbul Üniversitesi Adresli: Evet

Özet

 

Investıgatıon of the relatıonshıp of DHRS2 gene wıth ubıquıtın genes ın breast cancer

Damla Nur Şakar, Burcu Salman Yaylaz, Şeyma Punar, Sema Sırma Ekmekci, Neslihan Abacı

Purpose: Breast cancer is the most common malignancy in women and takes second place among all cancers. Interactions between p53 and Mdm2 molecules are widely researched, but these proteins’ regulatory mechanisms are still not fully known. The dehydrogenase/reductase gene we selected to study, DHRS2, regulates p53 via suppressing Mdm2. In addition, the ubiquitination process is commonly preserved during the evolution from yeast to human evolutions. Ubiquitins make many biochemical interactions in cell, and many different types of proteins (E1, E2 and E3) are included in regulating these interactions. Therefore, degradation of the misfolded or oncogenic proteins has importance in cancer treatment. In this study, we examine the ubiquitin genes as BARD1, BRCA2, STUB1, TRUSS, TRIM6, FBXW7, and BRCA1, during the expression changes of DHRS2, which is important for the cellular activity of breast cancer cell lines.

Methods: MCF10A, MCF7, T47D, and MDA MB 231 are four human breast cell lines used in this study. DHRS2 expressions were manipulated with gene specific siRNA and overexpression vectors with liposome-dependent transfection. After the transfection, total RNA isolation and cDNA synthesis occurred following the instructions. Quantification of the target genes made with qRT-PCR and calculated with ∆∆CT method. The highest DHRS2 silencing and overexpression were determined and used for analysis. The statistical comparison of the groups and the cells were made by Student’s t-test and Pearson correlation. Also, network analysis was used to determine the effect of DHRS2 gene expression on the ubiquitin genes..

Results: All the cells and both siDHRS2 and vekDHRS2 groups, TRUSS, STUB1 and MDM2 mRNA expressions were higher than non-transfected groups. On the other hand, TRIM6 expression decreased in all groups compared with NTC. FBXW7 expression was not changed in the MCF10A cell line. Luminal A type cell lines MCF7 and T47D cells showed similar gene expression levels except for MDM2 and BRCA1 genes. BRCA1 mRNA level was lower in both study groups than in NTC groups at T47D cells and higher in vekDHRS2 group at the MCF7 cell line. TRUSS gene expression was higher in triple negative cell line MDA MB 231 than in other cells. A high mRNA level of MDM2 in the vekDHRS2 group has a similar tendency with MCF7.

Conclusion: DHRS2, a short-chain dehydrogenase reductase enzyme, is known to regulate p53 degradation in cancer cells via an E3 ubiquitin ligase, MDM2. From this point of view, the effects of DHRS2 mRNA level change on other ubiquitin elements were investigated. It has been found that our target genes show different behavior due to malignancies of the cells and the oncogene mutations they carry. This study demonstrated that DHRS2 gene expression changes the expression of crucial intracellular ubiquitin genes. It has been a guide for further studies.

 


Disclosure Statement: The authors declare no conflict of interest