PhD Scientific Days 2020

Molecular Sciences IV. Posters

Alterations of DNA Methylation, DNA Repair, and Epithelial-mesenchymal Transition in Colorectal Cancer Cell Lines by S-Adenosylmethionine Treatment

Előadó neve

Dr. Zsigrai, Sára

Előadó munkahelye

Semmelweis University, 2nd Department of Internal Medicine, Molecular Gastroenterology Laboratory

Előadó telefonszáma

+36-1-266-0926/55622

Előadó e-mail címe

zsigrai.sara@gmail.com

Az előadás címe

Alterations of DNA Methylation, DNA Repair, and Epithelial-mesenchymal Transition in Colorectal Cancer Cell Lines by S-Adenosylmethionine Treatment

Szerző(k) neve és munkahelye

Sára Zsigrai1, Alexandra Kalmár1,2, Krisztina Andrea Szigeti1, Zsófia Brigitta Nagy1, Barbara Kinga Barták1, Gábor Valcz1,2, Orsolya Galamb1,2, Titanilla Dankó3, Zsolt Tulassay1,2, Péter Igaz1,2, Béla Molnár1,2
1 Semmelweis University, 2nd Department of Internal Medicine, Budapest, Hungary
2 Hungarian Academy of Sciences, Molecular Medicine Research Group, Budapest, Hungary
3 Semmelweis University, 1st Department of Pathology and Experimental Cancer Research, Budapest, Hungary

Szekció

Molecular Sciences IV. Posters

Language of the presentation

Hungarian

Section, first choice

Molecular Sciences

Section, second choice

Pathology and Oncology

Összefoglaló szövege

Introduction: S-adenosylmethionine (SAM) is a universal methyl donor molecule, used as a dietary supplement. SAM is involved in DNA methylation processes, thereby it may have a favorable effect on gene expression of cancer-associated genes through epigenetic modifications, but may also influence DNA folding during repair processes.

Aims: We aimed to analyze the effect of SAM treatment on global and promoter-specific DNA methylation levels, gene expression, DNA integrity, cell cycle, and the proliferation of two different colorectal cancer cell lines (HT-29, SW480).

Method: HT-29 and SW480 cells were treated with SAM in different concentrations (0, 0.5, 1 mmol/l) for 48 hours. Global DNA methylation status was analyzed by bisulfite pyrosequencing of long interspersed nuclear element-1 (LINE-1) retrotransposons. Promoter specific DNA methylation alterations were determined by Reduced Representation Bisulfite Sequencing (RRBS) method. Gene expression changes were detected using Human Transcriptome Array 2.0 (HTA 2.0). DNA integrity analysis was performed with γH2AX ELISA, immunostaining, and comet assay. Flow cytometry measurement and Sulforhodamine B (SRB) assay were assessed for cell cycle and proliferation determination.

Results: Global and promoter-specific DNA methylation alterations, as well as decreased expression (p< 0.05) of genes that are involved in epithelial-mesenchymal transition, were observed after SAM treatment. Increased phosphorylation of H2AX (74.9, 166.5, 200.6 pM) and decreased micronucleus number (1.47, 0.76, 0.45% of cells) were referred to the activation of reparative processes, that was supported by the changes of comet tale lengths. Treatment with SAM decreased the proportion of the cells in the G0/G1 phase (48.4, 28.5, 20.4%), while increased in the S (45.7, 61.7, 67.0%) and G2/M phases (6.0, 10.7, 12.5%). Significant (p< 0.05) reduction of cell proliferation (99.5, 77.97, 70.55%) was also detected with SRB assay.

Conclusion: SAM is able to alter the DNA methylation pattern of tumor cells and can induce DNA repair. Activation of these processes can lead to cell cycle arrest, decreased proliferation, and inhibition of epithelial-mesenchymal transition. Tumor cells could be targeted by SAM through different pathways; therefore, it may enhance the effect of chemotherapeutic agents.

Additional Information

Béla Molnár
bela.molnar1@med.semmelweis-univ.hu

Bemutatás módja

Poszter

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

4117

Start

12:52

End

12:55

Authors (legacy)

Sára Zsigrai1, Alexandra Kalmár1,2, Krisztina Andrea Szigeti1, Zsófia Brigitta Nagy1, Barbara Kinga Barták1, Gábor Valcz1,2, Orsolya Galamb1,2, Titanilla Dankó3, Zsolt Tulassay1,2, Péter Igaz1,2, Béla Molnár1,2
1 Semmelweis University, 2nd Department of Internal Medicine, Budapest, Hungary
2 Hungarian Academy of Sciences, Molecular Medicine Research Group, Budapest, Hungary
3 Semmelweis University, 1st Department of Pathology and Experimental Cancer Research, Budapest, Hungary