In Vitro Anticancer Activities of Senna singueana Leaf Extracts against HeLa Cell Lines

Document Type : Research Articles

Authors

1 Department of Biology, College of Natural and Computational Sciences, Aksum University, Axum, Ethiopia.

2 Department of Microbiology, GSL Medical College, Andhra Pradesh, India.

3 Department of Biochemistry, University College of Science and Technology, Adikavi Nannaya University, Raja mahendravaram, Andhra Pradesh, India.

4 School of Medicine, Texila American University, Lusaka, Zambia, Central Africa. Zambia.

5 Department of Chemistry, College of Natural and Computational Sciences, Aksum University, Axum, Ethiopia.

Abstract

Objective: The present demonstrated that SSL-CH extract selectively inhibits the growth of HeLa cell lines though activation of p53 and Bax genes medaied proapoptotic pathway signifying it is a potential alternative anticancer agent for cervical cancer. Materials and Methods: The selective cytotoxicity of diethyl ether, chloroform, ethyl acetate, and ethanolic leaf extracts of S. singueana were evaluated against cervical HeLa and normal mouse fibroblast L929 cell lines using MTT assay. At (IC50) concentration 20 µg/ml) of effective chloroform leaf extract of S. singueana (SSL-CH) was chosen to evaluate its nuclear apoptotic changes by using Acridine Orange/Etidium Bromide dual staining.  Subsequently, apoptotic induction potential of SSL-CH extract was assessed by RT-qPCR analysis of apoptotic genes p53 and Bax. Results: Treatment SSL-CH extract selectively inhibited the dose depended proflretion of HeLa cell lines with IC50 of 20 µg/ml. AO/EB dual staining showed SSL-CH extract induced late apoptosis of 49.42% in compared to the control. RT-PCR analyses demonstrated that SSL-CH extract profoundly upregulated the proapoptotic genes p53 to12.85 (***p≤0.01), Bax to 11.61 (**p≤0.01), caspase-9 to 40.62 (***p≤0.01), and caspase-3 to 11.61 folds (**p≤0.01) respectively and down regulated antiapoptotic genes Bcl-2 to 0.9 (**p≤0.001) and survivin gene to 1 (***p≤0.001) at 2×IC50 (40 µg/mL) in comparison to control, confirming activation of the intrinsic apoptotic pathway. Conclusion: These findings demonstrated that SSL-CH extract could serve as a potential alternative therapeutic agent for cervical cancer treatment with affordable and fewer side effects comparison with cisplatin.

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