Synergistic Cytotoxic Effect of Anonna muricata and Caesalpinia sappan Nanoparticles via Expression of BAD Pro-Apoptotic Protein in HeLa Cervical Cancer Cells

Okid P. Astirin¹, Adi Prayitno², Anif N. Artanti³, Elisa Herawati^1*, Anggraini R. Dewi¹, Virnanda R. Aryani¹, Vector S. Dewangga¹

¹Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University, Surakarta, Indonesia

²Department of Pathobiology, Faculty of Medicine, Sebelas Maret University, Surakarta, Indonesia

³Department of Pharmacy, Vocational College, Sebelas Maret University, Surakarta, Indonesia

Corresponding Author: [email protected]; Tel: +62-271-663375

Recieved Date: 20 October 2021; Accepted Date: 04 December 2021; Published Date: 04 January

Citation:

Astirin OP, Prayitno A, Artanti AN, Herawati E, Dewi AR, Aryani VR, Dewangga VS. Synergistic Cytotoxic Effect of Anonna muricata and Caesalpinia sappan Nanoparticles via Expression of BAD Pro-Apoptotic Protein in HeLa Cervical Cancer Cells. Trop J Nat Prod Res. 2021; 5(12): 2108-2114

http://www.doi.org/10.26538/tjnpr/v5i12.11

Copyright:

© 2021 Astirin et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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ABSTRACT

Annona muricata and Caesalpinia sappan are among the most important traditional medicinal plants, which contain numerous chemicals with various pharmacological properties. Although extracts from both plants have demonstrated anticancer activity, there has been no report on the anticancer effect of the combination of A. muricata and C. sappan nanoparticles particularly on cervical cancer (HeLa) cells. This study aimed to determine the optimum combination dose of both nanoparticles that showed a synergistic effect to induce apoptosis in HeLa cells. The nanoparticles were prepared using A. muricata leaves and C. sappan heartwood using the glass ionic method. Immunostaining was performed to evaluate the expression of pro-apoptotic marker BAD (BCL2 associated agonist of cell death) protein. Cytotoxicity effect was tested using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The size of the nanoparticles ranged between 237-453 nm and showed polydispersity index of 0.354 (A. muricata) and 0.486 (C. sappan), which means that the level of nanoparticle size distribution is quite uniform. BAD acts as an essential mediator of intrinsic apoptosis, as shown by enhanced expression of BAD in the group of cells treated with the nanoparticles. The employment of two nanoparticles dose combinations showed a synergistic cytotoxic effect (Combination Index<1) when each nanoparticle concentration was given at ½ IC50 (C. sappan) and ¼ IC50  (A. muricata), resulting in 52.51% of cell viability. The synergistic effect exhibited by A. muricata and C. sappan nanoparticles suggests a possible different target or signaling pathway that results in a reduction of required nanoparticles concentrations for individual sample.