AUTHOR=Lagu Inyani John Lino , Nyamai Dorothy Wavinya , Njeru Sospeter Ngoci 

TITLE=Phytochemical analysis, in-vitro and in-silico study of antiproliferative activity of ethyl acetate fraction of Launaea cornuta (Hochst. ex Oliv. & Hiern) C. Jeffrey against human cervical cancer cell line

JOURNAL=Frontiers in Pharmacology

VOLUME=Volume 15 - 2024

YEAR=2024

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2024.1399885

DOI=10.3389/fphar.2024.1399885

ISSN=1663-9812

ABSTRACT=Introduction: Cervical cancer is one of the leading causes of death among women globally due to the limitation of current
treatment methods and their associated adverse side effects Launaea cornuta (Hochst. ex Oliv. & Hiern) C. Jeffrey is used as
traditional medicine in the treatment of a variety of diseases, including cancer. Objective: This study aimed to evaluate the
selective antiproliferative, phytochemical profiles, and antimigratory effects of L. cornuta and to explore the modes of action in
human cervical cancer cell lines (HeLa-229). Materials and Methods: The cytotoxic effect of L. cornuta ethyl acetate fraction on the
proliferation of cervical cancer cells was evaluated by MTT assay. Compounds were analysed using the quantitative colour method
and gas chromatography-mass spectroscopy (GC-MS). Subsequently, bioinformatic analyses were performed to screen for potential
anticervical cancer and therapeutic target genes of L. cornuta. MD was performed to predict and understand the molecular
interactions and molecular mechanisms of the ligands against cervical cancer. A reverse transcription-quantitative polymerase
chain reaction was performed to validate the network analysis results. Results: L. cornuta ethyl acetate fraction exhibited a
remarkable cytotoxic effect on HeLa proliferation (IC50 of 20.56±2.83 µg/mL) with a selectivity index (SI) of 2.36 without affecting
Vero cells (IC50 of 48.83±23.02). Thirteen compounds were identified by GC-MS analysis. 124 potential target genes associated with
the effect of L. cornuta ethyl acetate fraction on human cervical cancer were obtained, including AKT1, MDM2, CDK2, MCL1 and
MTOR, identified among the top hub genes and PI3K/Akt1, and Ras/MAPK pathways identified as the significantly enriched
pathways. Molecular docking results showed that stigmasteryl methyl ether had a high binding affinity against CDK2, ATK1, BCL2,
MDM2, and Casp9, with binding energy ranging from -7.0 to 12.6 kcal/mol. Tremulone showed a good binding affinity against TP53
and P21 with -7.0 and 8.0 kcal/mol, respectively. RT-qPCR analysis revealed that CDK2, MDM2, and BCL2 were down-regulated, and
Casp9 and P21 were up-regulated in HeLa cells treated with L. cornuta compared to the negative control. Conclusions: The findings
indicate that L. cornuta ethyl acetate fraction phytochemicals targeting various targets and pathways exhibit selective cytotoxic
effects against HeLa cells.