Cui H et al. (2020), Antiproliferative Activity, Proapoptotic Effect...

ECB-ART-49440

Oxid Med Cell Longev 2020 Jun 26;2020:7948705. doi: 10.1155/2020/7948705.

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Antiproliferative Activity, Proapoptotic Effect, and Cell Cycle Arrest in Human Cancer Cells of Some Marine Natural Product Extract.

Cui H , Bashar MAE , Rady I , El-Naggar HA , Abd El-Maoula LM , Mehany ABM .

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Bioactive constituents of numerous marine organisms have been investigated recently for their preclinical and clinical anticancer activity. Three marine organisms: black-spotted sea cucumber: Pearsonothuria graeffei (Pg), lollyfish: Holothuria atra (Ha), and sea hare: Aplysia dactylomela (Ad), were collected during winter 2019 from Gulf of Aqaba, Red Sea, Egypt, and macerated with ethanol into three different extracts: PgE, HaE, and AdE, where each was in vitro assessed for its antiproliferative and proapoptotic properties on HepG2, HCT-116, and MCF-7 cancer cells. PgE dose-dependently inhibited the growth of HepG2, HCT-116, and MCF-7 cells within IC50 values 16.22, 13.34, and 18.09 μg/mL, respectively, while the IC50 values for the antiproliferative activity of HaE were 12.48, 10.45, and 10.36 μg/mL, respectively, and the IC50 values of AdE were 6.51, 5.33, and 6.87 μg/mL, respectively. All extracts were found to induce G0/G1 cell cycle arrest for HepG2 cells side by side with their inhibition of CDK2 on all three cell lines while all extracts were also showed to induce apoptosis in HepG2 cell line at pre-G 1 phase supplemented by their anticancer activity via proapoptotic protein Bax, caspase-3, and cleavage PARP increase, and antiapoptotic protein Bcl-2 downturn. Moreover, necrosis has been relatively noticed in HepG2 cell line as an additional anticancer activity for each extract. Our data introduced three ethanolic marine extracts as natural chemotherapeutic agents to be further developed for cancer control.

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	Figure 1. Antiproliferative activity of PgE, HaE, and AdE against HePG2, HCT-116, and MCF-7 cell lines.
	Figure 2. Cell cycle analysis effect in HepG2 cells treated with PgE, HaE, and AdE.
	Figure 3. Cell cycle analysis and apoptosis effect in HepG2 cell line treated with PgE, HaE, and AdE.
	Figure 4. Modulation of CDK2 protein expression in HePG2, HCT-116, and MCF-7 cells was treated dose-dependently with PgE, HaE, and AdE and harvested 24âh after treatments. The immunoblots shown are representative of three independent experiments which all gave similar results where only representative result was cropped and inserted here. Bars represent the means Â± SD. Single, double, triple, and quadruple asterisks are used for p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively, vs. control (DMSO-treated) cells.
	Figure 5. Comparative modulation of Bax, Bcl-2, and caspase-3 protein expressions in HePG2 cells was treated with PgE, HaE, and AdE and harvested 24âh after treatments.
	Figure 6. Modulation of the apoptotic protein biomarkers: PARP (116âkD) and cleaved PARP (85âkD) in HePG2, HCT-116, and MCF-7 cells, was treated dose-dependently with PgE, HaE, and AdE and harvested 24âh after treatments. The immunoblots shown are representative of three independent experiments which all gave similar results where the only representative result was cropped and inserted here. Bars represent the means Â± SD. Single, double, triple, and quadruple asterisks are used for p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively, vs. control (DMSO-treated) cells.
	Figure 7. Percentage of apoptosis and necrosis induced by PgE, HaE, and AdE in HepG2 cells.
	Figure 8. Cell death percentage induced by PgE, HaE, and AdE in HepG2 cells.
	Figure 9. Schematic drawing of the mechanism of action of PGE, HaE, and AdE on HepG2 cancer cells. This cartoon is based on the currently available data throughout the present study.
	Figure 10. Taxonomical positions and photographs of Pg, Ha, and Ad.