The effect of thymoquinone on cell proliferation, 8-hydroxy-2 -deoxyguanosine level and expression changes of DNA repair and oxidative stress-related genes in MCF-7 breast cancer cells

Abstract

Purpose: The aim of this study was to determine the effects of thymoquinone on cell proliferation, 8-hydroxy-2' -deoxyguanosine level and expression changes of oxidative stress and DNA repair-related genes in MCF-7 breast cancer cells. Material methods: Cell proliferation in MCF-7 cells after thymoquinone exposure was determined by MTT assay. 8-hydroxy-2' -deoxyguanosine protein concentration was measured by ELISA assay. Total RNA isolation from control and thymoquinone treated cell was performed by Trizol and cDNA was synthesized. mRNA expression changes of OGG1, NEIL-1, CRCC1 SOD2, CAT and NRF-2 were also determined in RT-PCR using SYBER Green method. Results: In this study, the IC50 dose of thymoquinone in MCF-7 cells was determined as 7.867 μM at 24th hour. It was investigated that thymoquinone inhibited 8-hydroxy-2' -deoxyguanosine level in breast cancer cells according to RT-PCR results, thymoquinone increased XRRC1 expression 4.71-fold and catalase expression 6.68-fold in breast cancer cells. Conclusion: In conclusion, TQ inhibits MCF-7 cell proliferation under in vitro conditions. It also alters the expression of genes associated with DNA repair and oxidative stress and acts through this oxidative stress mechanism. This study contributes to the existing literature and provides preliminary data for more comprehensive studies.

Keywords

• Thymoquinone
• MCF-7 cells
• breast cancer
• DNA repair genes
• oxidative stress

Timokinonun MCF-7 meme kanseri hücrelerinde hücre proliferasyonu, 8-hidroksi-2' –deoksiguanozin seviyesi ile DNA tamiri ve oksidatif stres ilişkili genlerin ekspresyon değişimlerine etkisi

Abstract

Amaç: Bu çalışmanın amacı, timokinonun MCF-7 meme kanseri hücrelerinde hücre proliferasyonu, 8-hidroksi-2' -deoksiguanozin düzeyi ve oksidatif stres ve DNA onarımı ile ilişkili genlerin ekspresyon değişiklikleri üzerindeki etkilerini belirlemektir. Gereç ve yöntem: Timokinon maruziyetinden sonra MCF-7 hücrelerinde hücre proliferasyonu MTT testi ile belirlendi. 8-hidroksi-2' -deoksiguanozin protein konsantrasyonu ELISA testi ile ölçüldü. Kontrol ve timokinon uygulanan hücrelerden Trizol ile total RNA izolasyonu yapıldı ve cDNA sentezlendi. OGG1, NEIL-1, CRCC1 SOD2, CAT ve NRF-2'nin mRNA ekspresyon değişiklikleri de SYBER Green yöntemi ile RT-PCR'da belirlendi. Bulgular: Bu çalışmada, MCF-7 hücrelerinde timokinonun IC50 dozu 24. saatte 7,867 μM olarak belirlenmiştir. Timokinonun meme kanseri hücrelerinde 8-hidroksi-2' -deoksiguanozin seviyesini inhibe ettiği araştırılmıştır. Ayrıca RT-PCR sonuçlarına göre, timokinon meme kanseri hücrelerinde XRRC1 ekspresyonunu 4,71 kat ve katalaz ekspresyonunu 6,68 kat artırmaktadır. Sonuç: Sonuç olarak, TQ in vitro koşullar altında MCF-7 hücre proliferasyonunu inhibe eder. Ayrıca DNA onarımı ve oksidatif stres ile ilişkili genlerin ekspresyonunu değiştirir ve bu oksidatif stres mekanizması yoluyla etki eder. Bu çalışma mevcut literatüre katkıda bulunmakta ve daha kapsamlı çalışmalar için ön veri sağlamaktadır.

Keywords

• Timokinon
• MCF-7 hücreleri
• meme kanseri
• DNA onarım genleri
• oksidatif stres

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