INVESTIGATION OF THE POSSIBLE EFFECT OF S-ALLYL-L-CYSTEINE ON APOPTOSIS AND AUTOPHAGY IN HUMAN LEUKEMIA CELL LINE

Year 2021, Volume: 22 Issue: 5, 373 - 380, 04.08.2021

Neslihan Tekin , Barış Kerimoğlu , Mehtap Tarhan Kamile Öztürk

https://doi.org/10.18229/kocatepetip.777181

Cited By: 1

Abstract

OBJECTIVE: S-Allyl-L-cysteine (SAC) is a biological active organosulfur component of garlic and has various pharmacological effects. SAC has displayed anti-cancer activity but the mechanism is unresolved. This study has focused on investigating the possible apoptotic and autophagic effects of SAC on two human leukemia cell lines: acute promyelocytic leukemia (HL-60) and chronic myeloid leukemia (K562).
MATERIAL AND METHODS: Cell cytotoxicity was evaluated via MTT test. Bax, Bcl-2, caspase 3, mTOR, AKT, and PI3K gene expression amounts were identified via Real time quantitative reverse transcription polymerase chain reaction (qRT-PCR). HL-60 and K562 cells were incubated with SAC at three diverse doses (5 mM, 10 mM, and 20 mM) (3,75 mM, 7,5 mM, and 15 mM), respectively.
RESULTS: SAC caused a cytotoxic effect on HL-60 and K562 cells with IC50 values of approximately 11.525 mM and 10.025 mM, respectively. In HL-60 cells, an increase in Bax expression levels was detected at doses of 5 mM and 10 mM SAC (p=0.027, p=0.000). Treatment with 10 mM SAC increased the expression level of caspase 3 in HL-60 cells as compared with the control and 5 mM SAC treated cells (p=0.000, p=0.020). In K562 cells, SAC induced a significant decrease in mTOR, AKT, and PI3K expression levels in at all doses (p=0.000, p=0.000, p=0.000).
CONCLUSIONS: In conclusion, our data indicates that SAC induces autophagy in K562 cells by downregulating the PI3K/AKT/mTOR signaling pathway. Furthermore, increased Bax and caspase 3 gene expression levels suggest that SAC may be an effective active ingredient with which to induce apoptosis in HL-60 cells.

Keywords

S-Allyl-L-cysteine, Bax, caspase 3, Bcl-2, mTOR

Supporting Institution

Aksaray Üniversitesi

Project Number

2015-076

Thanks

We are grateful to Aksaray University Science and Technology Application and Research Center for the use of the Molecular Biology and Metabolism Laboratory.

İNSAN LÖSEMİ HÜCRE HATLARINDA S-ALLİL-L-SİSTEİN’İN APOPTOZ VE OTOFAJİ ÜZERİNE OLASI ETKİLERİNİN ARAŞTIRILMASI

Abstract

AMAÇ: S-Allil-L-sistein (SAC), sarımsağın biyolojik olarak aktif bir organosülfür bileşenidir ve çeşitli farmakolojik etkilere sahiptir. SAC anti-kanser aktivite göstermektedir, ancak mekanizması belirlenememiştir. Bu çalışma, SAC'nin iki insan lösemi hücre dizisi üzerindeki olası apoptotik ve otofajik etkilerini araştırmaya odaklanmıştır: akut promiyelositik lösemi (HL-60) ve kronik miyeloid lösemi (K562).
GEREÇ VE YÖNTEM: Hücre sitotoksisitesi MTT testi ile değerlendirildi. Bax, Bcl-2, kaspaz 3, mTOR, AKT ve PI3K gen ekspresyon miktarları, kantitatif gerçek zamanlı ters transkripsiyon polimeraz zincir reaksiyonu (qRT-PCR) yoluyla tanımlandı. HL-60 ve K562 hücreleri, sırasıyla üç farklı dozda (5 mM, 10 mM ve 20 mM) (3,75 mM, 7,5 mM ve 15 mM) SAC ile inkübe edildi.
BULGULAR: SAC, sırasıyla yaklaşık 11.525 mM ve 10.025 mM IC50 değerleri ile HL-60 ve K562 hücreleri üzerinde sitotoksik etkiye neden olmuştur. HL-60 hücrelerinde, Bax ekspresyon seviyelerinde 5 mM ve 10 mM SAC dozlarında artış tespit edildi (p = 0.027, p = 0.000). 10 mM SAC ile tedavi, kontrol ve 5 mM SAC ile tedavi edilen hücrelere kıyasla HL-60 hücrelerinde kaspaz 3 ekspresyon seviyesini artırdı (p = 0.020, p = 0.000). K562 hücrelerinde SAC, tüm dozlarda mTOR, AKT ve PI3K ekspresyon seviyelerinde önemli bir düşüşe neden oldu (p = 0.000, p = 0.000, p = 0.000).
SONUÇ: Sonuç olarak, verilerimiz SAC'nin K562 hücrelerinde PI3K/AKT/mTOR sinyal yolunu down-regüle ederek otofajiyi indüklediğini göstermektedir. Ayrıca, artan Bax ve kaspaz 3 gen ekspresyon seviyeleri, SAC'nin HL-60 hücrelerinde apoptozu indüklemek için etkili bir aktif bileşen olabileceğini düşündürmektedir.

Keywords

S-Alil-L-sistein, Bax, kaspaz 3, Bcl-2, mTOR

Project Number

2015-076

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