Turmeric ethanol extract (<i>Curcuma longa</i> L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Alves, Naira Moura; Cruz, Vanessa de Sousa; Nepomuceno, Leandro Lopes; Soares, Nayane Peixoto; Arnhold, Emmanuel; Graziani, Daniel; Gonçalves, Patrícia de Almeida Machado; Badan, Guilherme Henrique Salazar; Santos, Amanda da Matta; Araújo, Eugênio Gonçalves de

doi:10.1590/1809-6891v23e-72215E

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Research article • Ciênc. anim. bras. 23 • 2022 • https://doi.org/10.1590/1809-6891v23e-72215E copy

Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 μM, 20 μM, 50 μM, 100 μM, and 1000 μM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC₅₀. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy, respectively. The extract enhanced the proliferation of canine osteosarcoma cells, reaching 3,819.74% at 50 μM of curcumin. The extract also significantly altered the expression of mutated p53 and Ki-67 proteins but not that of Bcl-2, suggesting that it did not induce this antiapoptotic pathway. Overall, these results are prerequisite to better understanding how natural compounds such as turmeric ethanolic extract affect cell proliferation and could be used to treat various diseases.

Keywords:
bone neoplasms; D-17; natural compounds; therapeutic safety

Curcumin Concentration (µM)	Semi-solid crude extract (mg)	Solution dilution
Curcumin Concentration (µM)	Semi-solid crude extract (mg)	DMSO (µL)	DMEM (µL)
0	0	4000	6000
10	6,0	4000	6000
20	12,0	4000	6000
50	31,0	4000	6000
100	59,0	4000	6000
1000	590,0	4000	6000

Exposure (h)	Concentration (µM)		Tukey
24 ^AB	0	100,00	d
	10	321,48	d
	20	625,28	cd
	50	3654,62	a
	100	2097,53	b
	1000	1262,41	c
48 ^A	0	100,00	d
	10	313,56	d
	20	297,94	d
	50	3945,09	a
	100	3000,44	b
	1000	1831,80	c
72 ^B	0	100,00	c
	10	197,88	c
	20	267,97	c
	50	3859,52	a
	100	2672,73	b
	1000	542,51	c

Curcumin Concentration (µM)	Cell Viability (%)	Tukey
0	100,00	d
10	277,65	d
20	397,07	d
50	3819,74	a
100	2590,23	b
1000	1212,24	c

Antibody	Curcumin Concentration(µM)	Posetively Labeled Cell Relation	Expression Intensity Score	Semiquantitative Combined Punctuation System
Bcl-2	0	5	2.1029^a	7.1029^a
	10	5	1.8750^a	6.8750^a
	20	5	1.4444^a	6.4444^a
	50	5	2.7500^a	7.7500^a
	100	5	2.5000^a	7.5000^a
	1000	5	2.2250^a	7.1000^a
p53	0	5	1.0441^b	6.0441^b
	10	5	1.4500^ab	6.4500^ab
	20	5	1.1111^b	6.1111^b
	50	5	1.6750^ab	6.6750^ab
	100	5	1.9000^ab	6.9000^ab
	1000	5	2.9000^a	7.9000^a
Ki-67	0	5	1.5588^ab	6.5588^ab
	10	5	0.8250^b	5.8250^b
	20	5	1.5000^ab	6.5000^ab
	50	5	2.2500^a	7.2500^a
	100	5	2.1500^ab	7.1500^ab
	1000	5	2.6000^a	7.6000^a

Universidade Federal de Goiás Universidade Federal de Goiás, Escola de Veterinária e Zootecnia, Campus II, Caixa Postal 131, CEP: 74001-970, Tel.: (55 62) 3521-1568, Fax: (55 62) 3521-1566 - Goiânia - GO - Brazil
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[1] *Corresponding author: earaujo@ufg.br