Potencial antitumoral e antimetastático de híbridos moleculares curcumina-cinamaldeído: estudos celulares e moleculares
Carregando...
Data
2020-02-20
Autores
Anselmo, Daiane Bertholin
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
O câncer é caracterizado por uma proliferação celular descontrolada, desencadeada por diversas mutações genéticas, fatores epigenéticos e ambientais. Apesar dos avanços terapêuticos, esta doença continua a ser responsável por índices de mortalidade elevados em todo o mundo, que é agravado pela grande resistência aos fármacos. Desta forma, se faz necessária a busca por novas substâncias potentes, seletivas e capazes de agir sobre células tumorais resistentes. Os produtos naturais têm despertado interesse científico devido suas inúmeras bioatividades. Dentre estes, a curcumina, isolada dos rizomas de Curcuma longa (popularmente conhecida como açafrão), o cinamaldeído, responsável pelo odor e sabor da canela (Cinnamomum sp.) e as chalconas, flavonoides abundantes em frutas, legumes e plantas, possuem potentes atividades antitumorais contra diversos tipos de câncer. O objetivo deste trabalho foi: a) avaliar a capacidade de inibição da proliferação celular de doze híbridos moleculares de curcumina-cinamaldeído contra células de câncer de mama MCF-7 e MDAMB-231, câncer de colo do útero C33, HeLa, SiHa e CaSki e câncer de vulva A431; b) selecionar o híbrido mais ativo e investigar os possíveis mecanismos celulares e moleculares em linhagens de osteossarcoma U2OS (p53 selvagem) e SAOS (p53 ausente). Pelo ensaio de MTT foi observado que os híbridos foram mais potentes que o cinamaldeído, sendo que híbridos hidroxilados foram mais ativos do que os híbridos metoxilados. O híbrido 5a apresentou maior potência antiproliferativa contra as células avaliadas de câncer feminino exceto para MCF-7, com valores de CI50 variando de 2,7 a 36,5 µM. O híbrido 5a demonstrou maior potência que a curcumina contra as células CaSki, SiHa, C33 e A431, exibiu uma maior seletividade que a curcumina, quando ambas foram testadas contra queratinócitos (HaCaT), além de ter demonstrado mais estável do que curcumina e cinamaldeído sob condições fisiológicas. Ensaios clonogênicos indicaram que 5a foi capaz de inibir a formação de colônias da linhagem A431, após 15 dias de tratamento. O híbrido 5a foi mais ativo contra células de osteossarcoma U2OS (CI50 = 22,4 µM) quando comparado às células SAOS (CI50 = 45,0 µM), sugerindo que a proteína p53 pode ser o seu alvo molecular. O híbrido 5a demonstrou boa atividade na inibição da migração e invasão de células U2OS por meio dos ensaios de wound-healing e transwell, respectivamente. Pela técnica de zimografia, o híbrido 5a indicou que efeitos sobre a migração e invasão pode estar relacionada com a diminuição da atividade da metaloproteinase de matriz 2 (MMP-2). A análise de Western Blot evidenciou a capacidade de 5a em induzir a expressão das proteínas p53 e DNAJB1 (Heat shock protein 40). Essa indução pode contribuir na atividade de p53, visto que Hsp40 estabiliza a conformação da proteína p53. As análises de citometria de fluxo indicaram que a inibição da viabilidade celular por 5a está relacionada com a indução de morte por apoptose, na qual p53 também pode estar envolvida. O estudo do potencial antitumoral e antimetastático de híbridos moleculares curcumina-cinamaldeídos é inédito na literatura, sendo o primeiro que avaliou a atividade de 5a em níveis celulares e moleculares.
Cancer is characterized by uncontrolled cell proliferation, triggered by several genetic mutations. Despite the therapeutic advances, this disease continues to be responsible for high mortality rates worldwide. This fact is aggravated by high drug resistance and combination therapy. Thus, it is necessary to search for new substances potent, selective and able to act on tumor cells resistant. Natural products have aroused scientific interest due to their innumerable bioavailabilities. Among these, curcumin, isolated from Curcuma longa rhizomes (popularly known as saffron) and cinnamaldehyde, responsible for the cinnamon odor and flavor (Cinnamomum sp.) and chalcones, flavonoids abundant in fruits, vegetables and plants, have potent anti-tumor activities against various cancers. The objective of this study was: a) evaluate the ability of inhibit cell proliferation of twelve curcumincinnamaldehyde molecular hybrids against MCF-7 and MDA-MB-231 breast cancer cells, C33, HeLa, SiHa and CaSki cervical cancer and A431 vulvar cancer; b) select the most active hybrid and investigate possible cellular and molecular mechanisms in osteosarcoma cell lines U2OS (p53 wild) and SAOS (p53 null). By the MTT assay it was observed that the hybrids were more potent than the cinnamaldehyde, with hydroxyl hybrids being more active than the methoxylated hybrids. Hybrid 5a presented higher antiproliferative potency against the cells evaluated for women cancer except for MCF-7, with IC50 values ranging from 2,7 to 36,5 μM. Hybrid 5a demonstrated higher potency than curcumin against CaSki, SiHa, C33 and A431 cells, and exhibited higher selectivity than curcumin when both were tested against keratinocytes (HaCaT), besides has show more stable than curcumin and cinnamaldehyde under physiological conditions. Clonogenic assays indicated that 5a was able to inhibit the colonies formation of A431 line after 15 days of treatment. Hybrid 5a was more active against U2OS osteosarcoma cells (IC50 = 22,4 μM) when compared to SAOS cells (IC50 = 45,0 μM), suggesting that the p53 protein may be its molecular target. Hybrid 5a demonstrated good activity in inhibiting the migration and invasion of U2OS cells by the wound-healing and transwell assays, respectively. By the zymography technique, hybrid 5a indicated that effects on migration and invasion may be related to decreased matrix metalloproteinase 2 (MMP-2) activity. Western Blot analysis demonstrated the ability of 5a to induce the expression of p53 and DNAJB1 (Heat shock protein 40). This induction may contribute to p53 activity, since Hsp40 stabilizes the conformation of the p53 protein. Flow cytometric analysis indicated that inhibition of cell viability by 5a is related to the induction of apoptosis death, in which p53 may also be involved. The study of the antitumor and antimetastatic potential of curcumincinnamaldehyde molecular hybrids is unprecedented in the literature, being the first to evaluate the activity of 5a at cellular and molecular levels.
Cancer is characterized by uncontrolled cell proliferation, triggered by several genetic mutations. Despite the therapeutic advances, this disease continues to be responsible for high mortality rates worldwide. This fact is aggravated by high drug resistance and combination therapy. Thus, it is necessary to search for new substances potent, selective and able to act on tumor cells resistant. Natural products have aroused scientific interest due to their innumerable bioavailabilities. Among these, curcumin, isolated from Curcuma longa rhizomes (popularly known as saffron) and cinnamaldehyde, responsible for the cinnamon odor and flavor (Cinnamomum sp.) and chalcones, flavonoids abundant in fruits, vegetables and plants, have potent anti-tumor activities against various cancers. The objective of this study was: a) evaluate the ability of inhibit cell proliferation of twelve curcumincinnamaldehyde molecular hybrids against MCF-7 and MDA-MB-231 breast cancer cells, C33, HeLa, SiHa and CaSki cervical cancer and A431 vulvar cancer; b) select the most active hybrid and investigate possible cellular and molecular mechanisms in osteosarcoma cell lines U2OS (p53 wild) and SAOS (p53 null). By the MTT assay it was observed that the hybrids were more potent than the cinnamaldehyde, with hydroxyl hybrids being more active than the methoxylated hybrids. Hybrid 5a presented higher antiproliferative potency against the cells evaluated for women cancer except for MCF-7, with IC50 values ranging from 2,7 to 36,5 μM. Hybrid 5a demonstrated higher potency than curcumin against CaSki, SiHa, C33 and A431 cells, and exhibited higher selectivity than curcumin when both were tested against keratinocytes (HaCaT), besides has show more stable than curcumin and cinnamaldehyde under physiological conditions. Clonogenic assays indicated that 5a was able to inhibit the colonies formation of A431 line after 15 days of treatment. Hybrid 5a was more active against U2OS osteosarcoma cells (IC50 = 22,4 μM) when compared to SAOS cells (IC50 = 45,0 μM), suggesting that the p53 protein may be its molecular target. Hybrid 5a demonstrated good activity in inhibiting the migration and invasion of U2OS cells by the wound-healing and transwell assays, respectively. By the zymography technique, hybrid 5a indicated that effects on migration and invasion may be related to decreased matrix metalloproteinase 2 (MMP-2) activity. Western Blot analysis demonstrated the ability of 5a to induce the expression of p53 and DNAJB1 (Heat shock protein 40). This induction may contribute to p53 activity, since Hsp40 stabilizes the conformation of the p53 protein. Flow cytometric analysis indicated that inhibition of cell viability by 5a is related to the induction of apoptosis death, in which p53 may also be involved. The study of the antitumor and antimetastatic potential of curcumincinnamaldehyde molecular hybrids is unprecedented in the literature, being the first to evaluate the activity of 5a at cellular and molecular levels.
Descrição
Palavras-chave
Câncer, p53, Apoptose, Curcumina, Cinamaldeído, Chalcona, Cancer, Apoptosis, Curcumin, Cinnamaldehyde, Chalcone