Britannin, a sesquiterpene lactone induces ROS-dependent apoptosis in NALM-6, REH, and JURKAT cell lines and produces a synergistic effect with vincristine.

Hassan Mohammadlou,Mohammad Hossein Mohammadi,Amir Yami,Maryam Hamzeloo-Moghadam,Ahmad Gharehbaghian

doi:10.1007/s11033-021-06572-x

Hassan Mohammadlou, Mohammad Hossein Mohammadi + Show 3 more

Open Access

https://doi.org/10.1007/s11033-021-06572-x

Copy DOI

Abstract

Britannin, a Sesquiterpene Lactone isolated from Inula aucheriana, has recently gained attraction in the therapeutic fields due to its anti-tumor properties. This study was designed to evaluate the effect of this agent on Acute Lymphoblastic Leukemia (ALL) cell lines, either as a monotherapy or in combination with Vincristine (VCR). To determine the anti-leukemic effects of Britannin on ALL-derived cell lines and suggest a mechanism of action for the agent, we used MTT assay, Annexin-V/PI staining, ROS assay, and real-time PCR analysis. Moreover, by using a combination index (CI), we evaluated the synergistic effect of Britannin on Vincristine. We found that unlike normal Peripheral Blood Mononuclear Cells (PBMCs) and L929 cells, Britannin reduced the viability of NALM-6, REH, and JURKAT cells. Among tested cells, NALM-6 cells had the highest sensitivity to Britannin, and this agent was able to induce p21/p27-mediated G1 cell cycle arrest and Reactive Oxygen Specious (ROS)-mediated apoptotic cell death in this cell line. When NALM-6 cells were treated with Nacetyl-L-Cysteine (NAC), a scavenger of ROS, Britannin could induce neither apoptosis nor reduce the survival of the cells suggesting that the cytotoxic effect of Britannin is induced through ROS-dependent manner. Moreover, we found that a low dose of Britannin enhanced the effect of Vincristine in NALM-6 cells by inducing apoptotic cell death via altering the expression of apoptotic-related genes. Overall, our results proposed a mechanism for the cytotoxic effect of Britannin, either as a single agent or in combination with Vincristine, in NALM-6 cells.

Highlights

As the most common malignancies in pediatrics, Acute Lymphoblastic Leukemia (ALL), with an incidence rate of 25% of all childhood malignancies is a heterogeneous disease that mostly affects children and teenagers aged between 1-19 years old [1, 2]
We found that when cells were exposed to NAC (80 μm), the ability of Britannin to reduce the metabolic activity of the cells was diminished (Fig. 4b), suggesting that probably Britannin induced its cytotoxic effects on NALM-6 cells through the production of Reactive Oxygen Specious (ROS)
We found that Britannin, a Sesquiterpene Lactone derived from Inula aucheriana, a native plant of West Azerbaijan (Iran), exerted cytotoxic effects on different ALL-derived cell lines such as Nalm-6, REH, and JURKAT

Summary

Introduction

As the most common malignancies in pediatrics, Acute Lymphoblastic Leukemia (ALL), with an incidence rate of 25% of all childhood malignancies is a heterogeneous disease that mostly affects children and teenagers aged between 1-19 years old [1, 2]. Despite the increase in event-free survival and the overall survival of patients (94.1%), the risk of disease relapse is one of the most challenging issues in the treatment of ALL [5]. Asteraceae plants are known for their biologically active sesquiterpene lactones Inula aucheriana, from this family, has been recently investigated for its chemical constituents, and Britannin, a Sesquiterpene Lactone (SL), has been reported from this plant [10, 11]. It has been shown that the SL compounds of this genus have cytotoxic effects due to the presence of α-methylene-γ-lactone [13]. It has shown that the SL compounds of this plant have anti-cancer effects due to the presence of α-methylene-γ-lactone [14] alongside their ability to treat infections and inflammatory diseases.

Objectives

Methods

Results

Conclusion