Amini E et al. (2017), In-vitro Pro Apoptotic Effect of Crude Saponin ...

ECB-ART-45471

Iran J Pharm Res 2017 Jan 01;161:266-276.

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In-vitro Pro Apoptotic Effect of Crude Saponin from Ophiocoma erinaceus against Cervical Cancer.

Amini E , Nabiuni M , Baharara J , Parivar K , Asili J .

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Ophiocoma erinaceus Muller &Troschel (Ophiocomidae) is part of the extensive group of echinoderm that contains bioactive metabolites. As the anti cancer potential of brittle star saponin has not been reported against cervical cancer, the present study was conducted to evaluate the anticancer effect of extracted crude saponin. Saponin extraction was conducted using conventional method such as froth test, TLC, FTIR and erythrolysis assay. The Hela-S3 cervical carcinoma and HNCF-PI52 normal cells were treated with different concentrations of saponin fraction for 24 and 48 h. The cytotoxicity was examined by MTT, DAPI, AO/PI, Annexin V-FITC and flow cytometry. In addition, the apoptotic induced pathway was studied using caspase assay, evaluation of ROS generation and Bcl-2 mRNA level. Crude saponin showed cytotoxic properties in Hela-S3 cells (IC50of 23.4 µg/mL) without significant impact against normal cells. In addition, the crude saponin increased sub-G1 peak in flow cytometry histogram of treated cells, ROS generation and caspase-3 and -9 activity (IC50 of 11.10, 11.27 µg/mL). The dose dependent down regulation of Bcl-2 in treated cells demonstrated that saponin fraction can trigger intrinsic apoptotic pathway in cancer cells. This study provides valuable information about the apoptotic inducing effect of saponin fraction, which can offer new insights into the anticancer potential of saponin as a promising candidate against human cervical carcinoma.

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Genes referenced: impact LOC115919910 ros1

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References [+] :

Andersson, Biological activity of saponins and saponin-like compounds from starfish and brittle-stars. 1989, Pubmed, Echinobase


	Figure 2. A) Fluorescence profiles of apoptosis induction by brittle star saponin fraction on cervical cancer cells using DAPI staining. (a-d) untreated cancer cells, treatment with 12.5, 25, 50 µg/ml brittle star saponin, respectively ×400. B) Fluorescence micrographs of brittle star saponin fraction on Hela-S3 cancer cells with AO/PI staining.(a-d) untreated cancer cells, treatment with 12.5, 25, 50 µg/ml brittle star saponin fraction. Magnification= ×200
	Figure 3. A) Evaluation of apoptosis by Annexin V-FITC method. The cells were exposed with increasing concentration of brittle star saponin fraction for 24 h by flow cytometry analysis.(a-d) untreated cancer cells, treatment with 12.5, 25 μg/ml brittle star saponin fraction, respectively. B) Apoptotic effect of brittle star saponin fraction on cervical cancer cells estimated by flow cytometry. Flow cytometry histogram of untreated and treated HeLa cells with 12.5, 25μg/ml brittle star saponin exhibited increase in sub-G1 region demonstrating mediation of an apoptotic cell death in cytotoxicity of brittle star saponin

	Figure 5. Effect of brittle star saponin fraction on intracellular ROS generation in Hela-S3 human cancer cells treated with brittle star saponin using fluorescence microscopy. A) untreated cells, B) 12.5 μg/ml, C) 25 μg/ml saponin treated cells. ×200