ISSN 1308-8734 | E-ISSN 1308-8742
Original Article
Investigation of the Anticancer Mechanism of Isoorientin Isolated from Eremurus Spectabilis Leaves via Cell Cycle Pathways in HT-29 Human Colorectal Adenocarcinoma Cells
1 Department of Physiology, Atatürk University School of Medicine, Erzurum, Turkey  
2 Department of Medical Biology, Pamukkale University School of Medicine, Denizli, Turkey  
3 Department of Pharmaceutical Botany, Atatürk University School of Pharmacy, Erzurum, Turkey  
Eurasian J Med 2018; 50: 168-172
DOI: 10.5152/eurasianjmed.2018.17403
Key Words: Isoorientin, colorectal cancer, cell cycle pathway
Abstract

Objective: Isoorientin (ISO) is a flavonoid compound extracted from plant species. The goal of this study was to determine the potential antiproliferative effects of ISO in HT-29 human colorectal adenocarcinoma cell line in vitro, specifically on cell viability, apoptosis, and cell cycle pathways.


Materials and Methods
: The cytotoxic effect of ISO isolated from E. spectabilis was measured using 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay in HT-29 cell lines. Total RNA was isolated using Tri-Reagent protocol. The effects of ISO on apoptosis-related gene were detected using real-time polymerase chain reaction (RT-PCR). The findings were analyzed using “Delta-Delta CT” ∆∆CT method and evaluated using a computer program. Volcano plot analysis was used for comparing groups and the data obtained were statistically analyzed using Student t test.


Results
: According to XTT result analysis, the 50% inhibitory concentration (IC50) value of ISO was 125 µM at the 48th h in HT-29 cells. The RT-PCR analysis in HT-29 cells showed that Cyclin D1 (CCND1 ), Cyclin-dependent kinase 6 (CDK6), BAX, BCL-2, Checkpoint kinase 1-2 (CHEK1, CHEK2) and Excision repair cross-complementing 1 (ERCC1) expressions were reduced in ISO-treated cells compared with those in the control group of cells. P53, P21, Caspase-3 (CASP-3), Caspase-8 (CASP-8), and Caspase-9 (CASP-9)  gene expressions were increased Ataxia Telengiectasia and Rad-3 related (ATR) was activated in the ISO-treated group of cells compared with those in the control group of cells (p<0.05).


Conclusion
: ISO affected the proliferation of colorectal cancer (CRC) cells via cell cycle pathways. It also altered apoptosis gene expression. These results demonstrated that ISO can be a therapeutic agent for CRC treatment; however, more studies are needed to investigate its mechanism of actions.


Cite this article as
: Gundogdu G, Dodurga Y, Elmas L, Yilmaz Tasci S, Karaoglan ES. Investigation of the anticancer mechanism of isoorientin isolated from Eremurus spectabilis leaves via cell cycle pathways in HT-29 human colorectal adenocarcinoma cells. Eurasian J Med 2018; 50(3): 168-172.

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