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Gene expression and anticancer evaluation of Kigelia africana (Lam.) Benth. Extracts using MDA-MB-231 and MCF-7 cell
Aasia Kalsoom1, Awais Altaf1, Huma Sattar1
1Institute of Molecular Biology (IMBB), Center for Research in Molecular Medicine (CRiMM), The University of Lahore, Lahore, Pakistan.
Abstract:
In recent years, a cancer research trend has shifted towards identifying novel therapeutic compounds from natural assets for the management of cancer. In this study, we aimed to assess the cytotoxic activity of Kigelia Africana (KA) extracts on breast cancer (MDA-MB-231 and MCF-7) and noncancerous kidney cells (HEK-293T) to develop an efficient anticancer medication. We used gas chromatography mass spectrometry (GC-MS to analyze the constituents of EKA and HKA extracts meanwhile the crystal violet and the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assays were used to examine the possible cytotoxic effects of plant extracts on our cancer cell lines along with non-cancerous control. The quantitative real-time PCR (RT-PCR) was run on cell samples to evaluate the differential expression of cell proliferative markers of cancer (BCL-2 and TP53). These phytochemicals have been reported to have binding affinity for some other growth factors and receptors as well which was evaluated by the in-silico molecular docking against Bcl2, EGFR, HER2, and TP53. Our Morphological observation showed a significant difference in the cell morphology and proliferation potential which was decreased under the effect of plant extracts treatment as compared to the control samples. The ethanol extract exhibited a marked antiproliferative activity towards MDA-MB-231 and MCF-7 cell lines with IC50 = 20 and 32 μg/mL, respectively. Quantitative RT-PCR gene expression investigation revealed that the IC50 concentration of ethanolic extract regulated the levels of mRNA expression of apoptotic genes. With the target and active binding site amino acids discovered in the molecular docking investigation, TP53/Propanoic acid, 3-(2, 3, 6-trimethyl-1, 4-dioxaspiro [4.4] non-7-yl)-, methyl ester (-7.1 kcal/mol) is the best-docked ligand. The use of this plant in folk remedies justifies its high in vitro anti-cancer capabilities. This work highlights the role of phytochemicals in the inhibition of cancer proliferation. Based on all these findings, it can be concluded that EKA extract has promising anti-proliferative effect on cancerous cells but more study is required in future to further narrow down the active ingredients of total crude extract with specific targets in cancer cells.
Insights
Kigelia Africana extracts show significant anticancer activity against breast cancer cells, inhibiting proliferation and regulating gene expression. Further research is needed to isolate specific active compounds for targeted cancer therapy.
Area of Science:
- Phytochemistry and Natural Products
- Cancer Biology and Therapeutics
- Molecular Pharmacology
Background:
- Growing interest in natural compounds for cancer treatment.
- Kigelia Africana (KA) is traditionally used in folk medicine.
- Need to scientifically validate the anticancer potential of KA extracts.
Purpose of the Study:
- To evaluate the cytotoxic and antiproliferative effects of Kigelia Africana extracts on breast cancer cell lines (MDA-MB-231 and MCF-7).
- To investigate the molecular mechanisms underlying the anticancer activity, including gene expression and molecular docking.
- To explore the potential of KA as a source for novel anticancer drug development.
Main Methods:
- Analysis of Kigelia Africana extracts using Gas Chromatography-Mass Spectrometry (GC-MS).
- Cytotoxicity assessment via Crystal Violet and MTT assays on breast cancer and non-cancerous kidney cells (HEK-293T).
- Gene expression analysis of proliferative markers (BCL-2, TP53) using quantitative RT-PCR.
- In-silico molecular docking against cancer-related proteins (Bcl2, EGFR, HER2, TP53).
Main Results:
- Ethanolic extract of Kigelia Africana (EKA) demonstrated significant antiproliferative activity with IC50 values of 20 μg/mL for MDA-MB-231 and 32 μg/mL for MCF-7 cells.
- Morphological changes and reduced proliferation were observed in cancer cells treated with KA extracts.
- EKA regulated the mRNA expression of apoptotic genes at IC50 concentrations and TP53 showed the best binding affinity with a docked ligand.
Conclusions:
- Kigelia Africana ethanolic extract exhibits promising in vitro anticancer properties against breast cancer cell lines.
- The study highlights the role of phytochemicals in inhibiting cancer cell proliferation and modulating gene expression.
- Further investigation is warranted to identify and characterize the specific active compounds within EKA for targeted cancer therapy development.
