In vitro antiproliferative effect and induction of apoptosis by Retama monosperma L. extract in human cervical cancer
N Merghoub1, L Benbacer, H El Btaouri
1Unité Biologie & Recherche Médicale, CNESTEN, Rabat, Maroc.
Abstract:
The antiproliferative effect of different extracts obtained from Retama monosperma L. was investigated on human SiHa and HeLa cervical cancer cell lines using a MTT colorimetric assay. The Retama monosperma L. dichloromethane fraction (Rm-DF) was the most active extract, exhibiting a significant cytotoxic activity on both cell lines in a dose-dependent manner, after 72 h of treatment. IC50 values obtained were 14.57 ± 4.15 μg/ml and 21.33 ± 7.88 μg/ml, for SiHa and HeLa cell lines respectively. The morphological features assessment of apoptosis in Rm-DF-treated cells showed a condensation of chromatin and apoptotic bodies, accompanied by a decrease in mitochondrial membrane potential (ΔΨm) and an increase in reactive oxygen species in both cell lines. The induction of apoptosis was further confirmed by Western blotting pro-caspase 3, Bcl2 and PARP; caspase 3 activity assay; and Annexin V labelling. Analysis of Rm-DF by CG/MS revealed the presence of five known quinolizidine alkaloids as well as, sparteine (10,97%), L-methyl cytisine (9.11%), 17-oxosparteine (3.49%), lupanine (0.93%) and anagyrine (39.63%). This study shows that Retama monosperma L. extract exhibits a potential anticancer activity against cervical cancer cell lines in vitro through the inhibition of proliferation and induction of apoptosis, which may involve a mitochondria-mediated signaling pathway.
Insights
Retama monosperma L. extract demonstrated significant antiproliferative and cytotoxic effects on cervical cancer cells. The study highlights its potential as an anticancer treatment by inducing apoptosis via a mitochondria-mediated pathway.
Area of Science:
- Phytochemistry
- Pharmacology
- Cancer Biology
Background:
- Cervical cancer remains a significant global health challenge.
- Natural products are a promising source for novel anticancer agents.
- Retama monosperma L. is a plant with potential medicinal properties.
Purpose of the Study:
- To investigate the antiproliferative and cytotoxic effects of Retama monosperma L. extracts on human cervical cancer cell lines (SiHa and HeLa).
- To elucidate the mechanism of action, focusing on apoptosis induction and mitochondrial pathways.
Main Methods:
- MTT colorimetric assay to determine cell viability and IC50 values.
- Morphological assessment of apoptosis, mitochondrial membrane potential (ΔΨm) analysis, and reactive oxygen species (ROS) detection.
- Western blotting for apoptosis-related proteins (pro-caspase 3, Bcl2, PARP), caspase 3 activity assay, and Annexin V labeling.
- Gas Chromatography/Mass Spectrometry (GC/MS) for chemical profiling of the active extract.
Main Results:
- The dichloromethane fraction (Rm-DF) of Retama monosperma L. showed significant dose-dependent cytotoxicity against both SiHa and HeLa cell lines (IC50 values: 14.57 ± 4.15 μg/ml and 21.33 ± 7.88 μg/ml, respectively).
- Rm-DF treatment induced morphological features of apoptosis, decreased mitochondrial membrane potential, and increased reactive oxygen species.
- Apoptosis induction was confirmed by Western blotting, caspase activity assays, and Annexin V staining.
- GC/MS analysis identified five quinolizidine alkaloids in Rm-DF, including anagyrine and sparteine.
Conclusions:
- Retama monosperma L. extract possesses significant in vitro anticancer activity against cervical cancer cell lines.
- The extract inhibits cancer cell proliferation and induces apoptosis, potentially through a mitochondria-mediated pathway.
- Quinolizidine alkaloids present in the extract may contribute to its observed anticancer effects, warranting further investigation.
