Browsing by Author "Kamau, Sally Wambui"

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    Network Pharmacology, Molecular Docking and Experimental Approaches of the Anti-Proliferative Effects of Rhamnus Prinoides Ethyl-Acetate Extract in Cervical Cancer Cells
    (Heliyon, 2024-09) Kamau, Sally Wambui; Ngugi, Mathew Piero; Mwikali, Peter Githaiga; Njeru, Sospeter Ngoci
    Background: Cervical cancer, one of the lethal cancers among women, is a challenging disease to treat. The current therapies often come with severe side effects and the risk of resistance development. Traditional herbal medicine, with its potential to offer effective and less toxic options, is a promising avenue. This study was undertaken to investigate the potential of Rhamnus prinoides (R. prinoides) root bark extracts in selectively inhibiting the proliferation of cervical cancer cells, using the HeLa cell line as an in vitro model. Methods: R. prinoides plant extracts were first screened at a fixed concentration of 200 μg/ml to determine the active extract. The selective anti-proliferative activity of the active extract was evaluated in a concentration dilution assay using the (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) MTT assay on cancerous (HeLa) cells and non-cancerous (Vero) cells to determine the half-maximal inhibitory (IC50) and half-cytotoxic concentrations (CC50), respectively. Functional assays on cell morphology (by microscopy), cell migration (wound healing assay) and cell cycle (by flow cytometry) were also conducted. The active extract was analyzed using Gas Chromatography/Mass Spectrometry (GC/MS) to determine any compounds it contained. Following identification of possible gene targets by network pharmacology, the genes were validated by molecular docking and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR). Results: The ethyl acetate extract of R. prinoides (EARP), the most active extract, selectively inhibited the growth of cervical cancer cells, their migration and induced cell cycle arrest at the S phase. In silico analysis revealed that squalene, 3,3a,6,6-tetramethyl-4,5,5a,7,8,9-hexahydro-1Hcyclopenta[i]indene and Olean-12-en-3.beta.-ol, acetate showed acceptable drug-like characteristics and may be partly attributed to the bioactivity demonstrated and the deregulation of the mRNA expression of AKT1, NF-κB, p53, Bax, Bcl-2, and Er-b-B2. Conclusion: This study, for the first time, demonstrates the anti-proliferation effects of EARP and forms a firm foundation for further drug development studies.
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    Phytochemical Profile, in Silico Molecular Docking Analysis and Anti-Cervical Cancer Effects of Rhamnus Prinoides and Grewia Villosa Extracts
    (Kenyatta University, 2025-07) Kamau, Sally Wambui
    Globally, cancer is the biggest cause of illness and mortality. Cancer comes second in prevalence in non-communicable disease in Kenya, after cardiovascular diseases. Cervical cancer is the leading cause of deaths in Kenya, resulting in as much as 11% of all cancer-related deaths. Presently there are several ways to treat cervical cancer: hysterectomy, radiation therapy, and chemotherapy. Chemotherapy is the most often utilized treatment because in Kenya, cervical cancer is typically diagnosed in its advanced stages. Although effective, chemotherapy is plagued by a myriad of challenges including severe side effects that greatly diminish the quality of living for the affected patients, the prohibitive cost of medical treatment and development of chemo-resistance to the chemotherapeutic drugs. Plant derived products are a feasible alternative in alleviating some of the challenges facing chemotherapy, especially in cervical cancer cases. Historically, Rhamnus prinoides and Grewia villosa plants have been utilized to cure and manage cancer and inflammatory illnesses. This study was undertaken to evaluate the phytochemical profile, the selective anti-proliferative activity of the extracts from R. prinoides and G. villosa root barks and their effects in limiting cell migration in vitro. In accordance with standard procedures, the 3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium (MTT) method was utilized to determine the anti-proliferative effects of the extracts in vitro, the in vitro scratch assay was used to determine the effects of the extracts in limiting cervical cancer cell migration while gas chromatography/mass spectrometry analysis was employed to identify extract specific compounds. The probable targets for the compounds found in the active extracts were identified using a variety of online databases and applications, and Pyrx software was utilized for molecular docking. R. prinoides ethyl acetate extract exhibited the most anti-proliferative effect having an IC50 value of 77.87 µg/ml while G. villosa ethyl acetate extract having an IC50 value of 100.70 µg/ml, similarly, exhibited the highest anti-proliferative effect among the extracts from G. villosa. The ethyl acetate extracts of both plants also had the highest selectivity indices with the R. prinoides extract having 4.40 and the G. villosa extract having a selectivity index of 2.48. The ethyl acetate extracts were also shown to inhibit cell migration in vitro with the IC50 concentration having the highest inhibitory effect after 48 hours. Phenols, triterpenoids, hydrocarbons, alkaloids, fatty acid esters were identified in the crude, hexane and ethyl acetate extracts of R. prinoides and G. villosa. 2,6,10-trimethyltetradecane and Benzene_1-methylundecyl compounds in the ethyl acetate extract of G. villosa and squalene, 3,3a,6,6-tetramethyl-4,5,5a,7,8,9-hexahydro-1H-cyclopenta[i]indene and Olean-12-en-3.beta.-ol,acetate compounds in the ethyl acetate extract of R. prinoides interacted with various oncogenic proteins with high binding affinity energies (<-4 kcal/mol). Network pharmacological analysis revealed that the compounds interacted with various proteins from key oncogenic pathways including the PI3K/Akt signaling pathway, carbon pathways in cancer and the EGFR tyrosine kinase resistance pathway. The ethyl acetate extracts of both plants upregulated TP53 mRNA levels while concurrently downregulating EGFR, ERBB2, and AKT1 mRNA levels. Additionally, the ethyl acetate extract of R. prinoides upregulated Bax mRNA levels while downregulating Bcl-2 and NF-kB mRNA levels, while the ethyl acetate extract of G. villosa upregulated Caspase 3 levels. In conclusion, the results from this study demonstrate the potential of Grewia villosa and Rhamnus prinoides extracts against cervical cancer in vitro and lay a solid foundation into further studies using the plant extracts for the development of drugs in cervical cancer therapy.