Astaxanthin Co-treatment with Low Dose Methotrexate Increases the CellCycle Arrest and Ameliorates the Methotrexate-induced InflammatoryResponse in NALM-6

Document Type : Original Article

Authors
Nastaran Moridi 1
Mahsa Najafzadeh 1
Seyed Mehdi Sajjadi 2
Mahtab Sayedi 3
1 Student Research Committee, Birjand University of Medical Sciences, Birjand, Iran.
2 Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran.
3 Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran
10.22088/IJMCM.BUMS.13.2.133
Abstract
Methotrexate (MTX), an antimetabolite agent, is widely used for acute lymphoblastic leukemia
treatment, despite its association with significant organ dysfunction. Astaxanthin (AST) is a natural
carotenoid which has recently been emerged as a promising anti-tumor and anti-inflammatory agent.
In this study, we aimed to evaluate the effectiveness of astaxanthin and low-dose methotrexate co-
treatment in acute lymphoblastic leukemia cell line. The expression of Dihydrofolate reductase
(DHFR), Thymidylate synthase (TYMS), apoptotic, anti-apoptotic as well as inflammatory genes was
investigated using qRT-PCR. Flow cytometry was performed for cell cycle quantitative evaluation.
Clonogenic assay was used to assess NALM6 cells proliferation capacity following treatment with
AST, MTX, and co-treatment. To compare the antioxidant property of each group, the ferric ion
reducing anti-oxidant power assay was performed. A reduction in viability was observed in the
presence of MTX, AST, and their combined treatment. Both AST alone and in combination with MTX
caused cell cycle arrest and a reduction in the expression of DHFR and TYMS. While MTX, AST,
and their combination could reduce STAT3 and BCL-XL gene expression, they could act as positive
regulators for the expression of BAX and CASP3, TNFα, and IL6. AST and MTX co-treatment
inhibited the colony formation ability. FRAP assay also revealed that AST and AST+MTX increased
the antioxidant capacity. Our data suggests that AST can improve MTX treatment efficacy and their
combination therapy can be considered as a promising strategy for the management of acute
lymphoblastic leukemia.
Keywords
Acute lymphoblastic leukemia
Astaxanthin
Methotrexate
NALM-6