Retama monosperma n-hexane extract induces cell cycle arrest and extrinsic pathway-dependent apoptosis in Jurkat cells

Lamiae Belayachi,Youssef Bakri,Priam Villalonga,Silvia Fernández De Mattos,Nawel Merghoub,Saaïd Amzazi,Clara Aceves-Luquero

doi:10.1186/1472-6882-14-38

Abstract

BackgroundRetama monosperma L. (Boiss.) or Genista monosperma L. (Lam.), locally named as “R’tam”, is an annual and spontaneous plant belonging to the Fabaceae family. In Morocco, Retama genus is located in desert regions and across the Middle Atlas and it has been widely used in traditional medicine in many countries. In this study, we show that Retama monosperma hexane extract presents significant anti-leukemic effects against human Jurkat cells.MethodsHuman Jurkat cells, together with other cell lines were screened with different concentrations of Retama monosperma hexane extract at different time intervals. Growth inhibition was determined using luminescent-based viability assays. Cell cycle arrest and apoptosis were measured by flow cytometry analysis. Combined caspase 3 and 7 activities were measured using luminometric caspase assays and immunoblots were performed to analyze expression of relevant pro- and anti-apoptotic proteins. GC-MS were used to determine the chemical constituents of the active extract.ResultsRetama monosperma hexane extract (Rm-HE) showed significant cytotoxicity against Jurkat cells, whereas it proved to be essentially ineffective against both normal mouse fibroblasts (NIH3T3) and normal lymphocytes (TK-6). Cytometric analysis indicated that Rm-HE promoted cell cycle arrest and apoptosis induction accompanied by DNA damage induction indicated by an increase in p-H2A.X levels. Rm-HE induced apoptosis was partially JNK-dependent and characterized by an increase in Fas-L levels together with activation of caspases 8, 3, 7 and 9, whereas neither the pro-apoptotic nor anti-apoptotic mitochondrial membrane proteins analyzed were significantly altered. Chemical identification analysis indicated that α-linolenic acid, campesterol, stigmasterol and sitosterol were the major bioactive components within the extract.ConclusionsOur data suggest that bioactive compounds present in Rm-HE show significant anti leukemic activity inducing cell cycle arrest and cell death that operates, at least partially, through the extrinsic apoptosis pathway.

Highlights

Retama monosperma L. (Boiss.) or Genista monosperma L. (Lam.), locally named as “R’tam”, is an annual and spontaneous plant belonging to the Fabaceae family
We describe the major cellular effects of Retama monosperma hexane extract (Rm-HE) leading to cell cycle arrest and extrinsic pathway-dependent apoptosis
Analysis of the cytotoxic activity of Rm-HE against human cancer cell lines To investigate the potential effect of Rm-HE extract against cancer cells, various human cancer cell lines of different origin were screened to assess the cytotoxic activity of Rm-HE

Summary

Introduction

Retama monosperma L. (Boiss.) or Genista monosperma L. (Lam.), locally named as “R’tam”, is an annual and spontaneous plant belonging to the Fabaceae family. In Morocco, Retama genus is located in desert regions and across the Middle Atlas and it has been widely used in traditional medicine in many countries. In Morocco, Retama genus is largely located in desert regions and the Middle Atlas [4]. This plant is used in traditional medicine in many countries, as purgative, vermifuge, antihelmintic, and abortive [5]. Several studies have investigated Retama genus for various pharmacological effects, including hypoglycemic and diuretic [4,6], cytotoxic [7,8], antioxidant, antiviral [9], antihypertensive [10], antiinflammatory [11] and antitumor activities [12]

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