Malyarenko OS et al. (2021), Combined Anticancer Effect of Sulfated Laminara...

ECB-ART-48906

Mar Drugs 2021 Sep 25;1910:. doi: 10.3390/md19100540.

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Combined Anticancer Effect of Sulfated Laminaran from the Brown Alga Alaria angusta and Polyhydroxysteroid Glycosides from the Starfish Protoreaster lincki on 3D Colorectal Carcinoma HCT 116 Cell Line.

Malyarenko OS , Malyarenko TV , Usoltseva RV , Surits VV , Kicha AA , Ivanchina NV , Ermakova SP .

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Colorectal cancer is one of the most frequent types of malignancy in the world. The search for new approaches of increasing the efficacy of cancer therapy is relevant. This work was aimed to study individual, combined anticancer effects, and molecular mechanism of action of sulfated laminaran AaLs of the brown alga Alaria angusta and protolinckiosides A (PL1), B (PL2), and linckoside L1 (L1) of the starfish Protoreaster lincki using a 3D cell culture model. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS), soft agar, 3D spheroids invasion, and Western blotting assays were performed to determine the effect and mechanism of the action of investigated compounds or their combinations on proliferation, colony formation, and the invasion of 3D HCT 116 spheroids. AaLs, PL1, PL2, and L1 individually inhibited viability, colony growth, and the invasion of 3D HCT 116 spheroids in a variable degree with greater activity of linckoside L1. AaLs in combination with L1 exerted synergism of a combined anticancer effect through the inactivation of protein kinase B (AKT) kinase and, consequently, the induction of apoptosis via the regulation of proapoptotic/antiapoptotic proteins balance. The obtained data about the efficacy of the combined anticancer effect of a laminaran derivative of brown algae and polyhydroxysteroid glycosides of starfish open up prospects for the development of new therapeutic approaches for colorectal cancer treatment.

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	Figure 1. The structures of polyhydroxysteroid glycosides PL1, PL2, and L1 from P. lincki.
	Figure 2. Individual effect of PL1, PL2, L1, and AaLs on viability and proliferation of 3D HCT 116 spheroids. The 3D HCT 116 spheroids were treated with (A) PL1, (B) PL2, (C) L1 (6.25–200 μM), and (D) AaLs (400–1600 μg/mL) for 24 h, 48 h, and 72 h. Cell viability was assessed using the MTS assay. All analyses were performed in three independent experiments. Results are expressed as mean ± standard deviation (SD).
	Figure 3. Combined effect of PL1, PL2, L1, with AaLs on viability and proliferation of 3D HCT 116 spheroids. The 3D HCT 116 spheroids were treated with PL1 (A), PL2 (B), and L1 (C) (12.5 μM) in combination with AaLs (10, 20, and 40 μg/mL) for 72 h. Cell viability was assessed using the MTS assay. Photos (n = 6 for control or cells treated by PL1, PL2, or L1 with AaLs, where n = the number of photos) of each spheroid were made using a ZOE™ Fluorescent Cell Imager and the analysis of spheroids was conducted using ImageJ software bundled with 64-bit Java 1.8.0_112 (NIH, Bethesda, Maryland, USA)). (D) Type of combination of L1 with AaLs calculated by Compusyn software 1.0.1 (ComboSyn, Inc., Paramus, NJ, USA). Combination index (CI) is a quantitative measure of the degree of interaction between different treatments. A CI equal to 0.9–1.1 is considered additive; a CI value of greater than 1.1 represents antagonism; and CI values less than 0.7 denotes synergism. All analyses were performed in three independent experiments. Results are expressed as mean ± standard deviation (SD). The Student’s t test was used to evaluate data with the following significance level *** p < 0.001.
	Figure 4. Individual and combined effects of polyhydroxysteroid glycosides from P. lincki and sulfated derivative of laminaran from A. angusta on the colony growth of 3D HCT 116 spheroids. The 3D HCT 116 spheroids were treated with (A) PL1, PL2, L1 (12.5, 25, 50 μM); (B) AaLs (100, 200, 400 μg/mL) or (C,D) PL1 (12.5 μM) in combination with AaLs (10, 20, 40 μg/mL); (E,F) PL2 (12.5 μM) in combination with AaLs (10, 20, 40 μg/mL); and (G,H) L1 (12.5 μM) in combination with AaLs (10, 20, 40 μg/mL) in soft agar. Photos of colonies were made using a Motic microscope (40× magnification, 500 μm scale). Results are expressed as mean ± standard deviation (SD). The Student’s t test was used to evaluate data with the following levels of significance: * p < 0.05, p < 0.01, * p < 0.001. All analyses were performed in three independent experiments. (J) The Combination index (CI) calculated using the Compusyn software 1.0.1 (ComboSyn, Inc., Paramus, NJ, USA), is a quantitative measure of the interaction between L1 and AaLs in a 3D cell culture model. CI of less than 0.7 indicates a synergism of the effect of L1 and AaLs.
	Figure 5. Individual effects of polyhydroxysteroid glycosides from P. lincki and sulfated derivative of laminaran from A. angusta on the invasion of 3D HCT 116 spheroids. The 3D HCT 116 spheroids were treated with (A) PBS (control) (B) PL1, (C) PL2, (D) L1 (12.5, 25, 50 μM) or (E) AaLs (200, 400, 800 μg/mL) after 72 h of invasion into the ECM gel. Photos of spheroids were made using a Motic microscope (40× magnification, 500 μm scale). (F) Quantitative measurement of the invasion area of spheroids treated with individual compounds was carried out using the ImageJ program and determined as the difference between total area invaded by cells leaving the spheroid and the area of the spheroid. All analyses were performed in three independent experiments. Results are expressed as mean ± standard deviation (SD). The Student’s t test was used to evaluate data with the following levels of significance *** p < 0.001.
	Figure 6. Combined effects of polyhydroxysteroid glycosides from P. lincki and sulfated derivative of laminaran from A. angusta on the invasion of 3D HCT 116 spheroids. (A) Photos (40× magnification, 500 μm scale) of 3D HCT 116 spheroids were treated with L1 (12.5 μM) in combination with AaLs (10, 20, 40 μg/mL) after 72 h invasion into the ECM gel. (B) Quantification of the invasion area of the spheroids was evaluted with the aid of ImageJ program. (C) The CI, calculated using the Compusyn software 1.0.1 (ComboSyn, Inc., Paramus, NJ, USA), is a quantitative measure of the interaction between L1 and AaLs in a 3D spheroids invasion model. (D) The regulation of the activity of AKT (Ser 473) kinase and apoptotic proteins Bcl-XL, Bax, caspase 3, and cleaved caspase 3 by L1 in combination with AaLs. (E) Relative band density was measured using Image Lab™ Software 4.1. Results are expressed as mean ± standard deviation (SD). The Student’s t test was used to evaluate data with the following levels of significance: p < 0.01, * p < 0.001.

References [+] :

Bae, Laminarin-Derived from Brown Algae Suppresses the Growth of Ovarian Cancer Cells via Mitochondrial Dysfunction and ER Stress. 2020, Pubmed