RESUMO
The CopA3 dimer peptide is a coprisin analog that has an anticancer effect against human cancer cells in vitro. In this study, we investigated the anticancer activity of the enantiomeric CopA3 dimer peptide in human gastric cancer cell lines as well as in an in vivo tumor xenograft model. Enantiomeric CopA3 reduced gastric cancer cell viability and exhibited cytotoxicity against cancer cells. Enantiomeric CopA3-induced cell death was mediated by specific interactions with phosphatidylserine and phosphatidylcholine, membrane components that are enriched in cancer cells, in a calcein leakage assay. Moreover, acridine orange/ethidium bromide staining, flow cytometric analysis, and Western blot analysis showed that enantiomeric CopA3 induced apoptotic and necrotic gastric cancer cell death. The antitumor effect was also observed in a mouse tumor xenograft model in which intratumoral inoculation of the peptide resulted in a significant decrease in the SNU-668 gastric cancer tumor volume. In addition, periodic acid-Schiff and hematoxylin staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay revealed apoptotic and necrotic cell death in tumor masses treated with greater than 150 µg CopA3. Collectively, these results indicate that the enantiomeric CopA3 dimer peptide induces apoptosis and necrosis of gastric cancer cells in vitro and in vivo, indicating that the peptide is a potential candidate for the treatment of gastric cancer, which is a common cause of cancer and cancer deaths worldwide.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Proteínas de Insetos/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Ensaios Antitumorais Modelo de Xenoenxerto , Sequência de Aminoácidos , Animais , Peptídeos Catiônicos Antimicrobianos/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Caspases/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HeLa , Humanos , Proteínas de Insetos/química , Camundongos Endogâmicos BALB C , Camundongos Nus , Multimerização Proteica , Transdução de Sinais/efeitos dos fármacos , Estereoisomerismo , Neoplasias Gástricas/patologia , Carga Tumoral/efeitos dos fármacosRESUMO
Previously, we performed de novo RNA sequencing of Scolopendra subspinipes mutilans using high-throughput sequencing technology and identified several antimicrobial peptide candidates. Among them, a cationic antimicrobial peptide, scolopendrasin VII, was selected based on its physicochemical properties, such as length, charge, and isoelectric point. Here, we assessed the anticancer activities of scolopendrasin VII against U937 and Jurkat leukemia cell lines. The results showed that scolopendrasin VII decreased the viability of the leukemia cells in MTS assays. Furthermore, flow cytometric analysis and acridine orange/ethidium bromide staining revealed that scolopendrasin VII induced necrosis in the leukemia cells. Scolopendrasin VII-induced necrosis was mediated by specific interaction with phosphatidylserine, which is enriched in the membrane of cancer cells. Taken together, these data indicated that scolopendrasin VII induced necrotic cell death in leukemia cells, probably through interaction with phosphatidylserine. The results provide a useful anticancer peptide candidate and an efficient strategy for new anticancer peptide development.
Assuntos
Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/farmacologia , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Linfócitos B/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Animais , Linfócitos B/fisiologia , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Alcaloides Diterpenos , Humanos , Células Jurkat , Fosfatidilserinas/metabolismo , Células U937RESUMO
In this study, the compositions of Ophiocordyceps longissima mycelia, synnemata, and fruiting bodies were first analyzed in order to clarify its chemical basis for development as a health food or medicine. We found that the contents of crude protein, polysaccharides, and macroelements were highest in mycelia, whereas effective components, including mannitol, ergosterol, adenosine, inosine, Zn, and Se, were lowest in mycelia. Polysaccharide, mannitol, and ergosterol levels in synnemata (2.33, 4.54, and 0.66 g/100 g, respectively) were similar to those in fruiting bodies, but was significantly different from those of mycelia (4.79, 1.77, and 0.43 g/100 g). Trehalose content in fruiting bodies (2.15 g/100 g) was >4 times higher than that in synnemata (0.5 g/100 g). Adenosine content in fruiting bodies (0.024 g/100 g) was 3-4 times higher than that of synnemata, whereas inosine, cytosine, guanosine, and uridine (0.093, 0.145, 0.053, and 0.073 g/100 g) were highest in synnemata. Cu and Se were lower in mycelia (3.30 × 10â»4 and 3.1 × 10â»5 g/100 g) than in synnemata (1.85 × 10⻳ and 7.2 × 10â»5 g/100 g) and fruiting bodies (1.67 × 10⻳ and 4.3 × 10â»5 g/100 g). As, Hg, Cd, and Pb in mycelia were under the limit of edible fungus health standard; Pb was not found, but Cd, Hg, and As were detected in fruiting bodies. These findings suggest that O. longissima could be utilized in different culture methods according to market demand and might be a possible health food or medicinal resource.
Assuntos
Hypocreales/química , Extratos Vegetais/análise , Cobre/análise , Carpóforos/química , Carpóforos/crescimento & desenvolvimento , Hypocreales/crescimento & desenvolvimento , Micélio/química , Micélio/crescimento & desenvolvimento , Valor Nutritivo , Proteínas de Plantas/análise , Polissacarídeos/análiseRESUMO
CopA3 is a homodimeric α-helical peptide derived from coprisin which is a defensin-like antimicrobial peptide that was identified from the dung beetle, Copris tripartitus. CopA3 has been reported to have anticancer activity against leukemia cancer cells. In the present study, we investigated the anticancer activity of CopA3 in human gastric cancer cells. CopA3 reduced cell viability and it was cytotoxic to gastric cancer cells in the MTS and LDH release assay, respectively. CopA3 was shown to induce necrotic cell death of the gastric cancer cells by flow cytometric analysis and acridine orange/ethidium bromide staining. CopA3-induced cell death was mediated by specific interactions with phosphatidylserine, a membrane component of cancer cells. Taken together, these data indicated that CopA3 mainly caused necrosis of gastric cancer cells, probably through interactions with phosphatidylserine, which suggests the potential utility of CopA3 as a cancer therapeutic.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Antineoplásicos/farmacologia , Proteínas de Insetos/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células HeLa , Humanos , Camundongos , Fosfatidilserinas/metabolismo , Células RAW 264.7 , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologiaRESUMO
Harmoniasin is a defensin-like antimicrobial peptide identified from the ladybug Harmonia axyridis. Among the synthetic homodimer peptide analogues derived from harmoniasin, HaA4 has been found to have antibacterial activity without hemolytic activity. In this study, we investigated whether HaA4 has anticancer activity against human leukemia cell lines such as U937 and Jurkat cells. HaA4 manifested cytotoxicity and decreased the cell viability of U937 and Jurkat cells in MTS assay and LDH release assay. We found that HaA4 induced apoptotic and necrotic cell death of the leukemia cells using flow cytometric analysis, acridine orange/ethidium bromide staining and nucleosomal fragmentation of genomic DNA. Activation of caspase-7 and -9 and fragmentation of poly (ADP-ribose) polymerase was detected in the HaA4-treated leukemia cells, suggesting induction of a caspase-dependent apoptosis pathway by HaA4. Caspase-dependent apoptosis was further confirmed by reversal of the HaA4-induced viability reduction by treatment of Z-VAD-FMK, a pan-caspase inhibitor. In conclusion, HaA4 caused necrosis and caspase-dependent apoptosis in both U937 and Jurkat leukemia cells, which suggests potential utility of HaA4 as a cancer therapeutic agent.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Proteínas de Insetos/farmacologia , Animais , Peptídeos Catiônicos Antimicrobianos/química , Caspase 7/efeitos dos fármacos , Caspase 7/metabolismo , Caspase 9/efeitos dos fármacos , Caspase 9/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Humanos , Células Jurkat , Leucemia/tratamento farmacológico , Camundongos , Poli(ADP-Ribose) Polimerases/efeitos dos fármacos , Poli(ADP-Ribose) Polimerases/metabolismo , Células U937RESUMO
Our previous study demonstrated that CopA3, a disulfide dimer of the coprisin peptide analogue (LLCIALRKK), has antibacterial activity. In this study, we assessed whether CopA3 caused cellular toxicity in various mammalian cell lines. CopA3 selectively caused a marked decrease in cell viability in Jurkat T, U937, and AML-2 cells (human leukemia cells), but was not cytotoxic to Caki or Hela cells. Fragmentation of DNA, a marker of apoptosis, was also confirmed in the leukemia cell lines, but not in the other cells. CopA3-induced apoptosis in leukemia cells was mediated by apoptosis inducing factor (AIF), indicating induction of a caspase-independent signaling pathway.
Assuntos
Apoptose/efeitos dos fármacos , Caspases/metabolismo , Proteínas de Insetos/toxicidade , Sequência de Aminoácidos , Animais , Fator de Indução de Apoptose/metabolismo , Linhagem Celular Tumoral , Besouros/metabolismo , Células HeLa , Humanos , Proteínas de Insetos/síntese química , Proteínas de Insetos/uso terapêutico , Células Jurkat , Leucemia/tratamento farmacológico , Leucemia/metabolismo , Transdução de SinaisRESUMO
Paecilomyces tenuipes reportedly have anticancer and immune activities, along with various other medicinal uses. Cultured products with P. tenuipes are certified for use in food in South Korea, and processed goods containing this fungus have been developed in many countries, particularly South Korea, Japan, and China. Research on mass production technology-procured raw materials for the manufacture of P. tenuipes is very important; however, cultures of the fungus have been unstable. This study identified stable cultivation conditions, focusing on growth inhibition and revitalization. Moisture regulation and preservation of pupae inoculated with P. tenuipes were used to control growth inhibition and revitalization. When inoculated silkworm pupae were dehydrated to 4% moisture and preserved freeze-dried or at -70 degrees C, -20 degrees C, or 4 degrees C, the mycelia in their bodies were able to survive for 14 d. Inoculated silkworm pupae were rehydrated for 3 h and the mycelia within their bodies were recovered at 94.3-96.3%. Silkworm pupae at 4% moisture were able to survive for 135 d at temperatures < 4 degrees C and for 1 y after freeze-drying. Optimal conditions for synnemata induction were 25 degrees C and 100-300 1x.
Assuntos
Bombyx/microbiologia , Carpóforos/crescimento & desenvolvimento , Micélio/crescimento & desenvolvimento , Paecilomyces/crescimento & desenvolvimento , Preservação Biológica/métodos , Animais , Carpóforos/efeitos da radiação , Larva/microbiologia , Luz , Paecilomyces/isolamento & purificação , Paecilomyces/efeitos da radiação , Pupa/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , TemperaturaRESUMO
The aim of the present study was to investigate the anti-tumor effects of a culture filtrate of Paecilomyces farinosus J3. Various anti-tumor assays using B16 melanoma cells were carried out. Paecilomyces farinosus J3 significantly decreased the wound healing capability, invasiveness and angiogenic activity, which was confirmed by wound healing, human umbilical vein endothelial cell and invasion assays. Paecilomyces farinosus J3 strongly inhibited cell migration, tube formation and the angiogenic process in a concentration-dependent manner. Zymographic analysis also indicated a reduced expression of matrix metalloproteinase-9 (MMP-9), a 92-kDa gelatinase. Taken together, the results indicate that the anti-tumor activities of Paecilomyces farinosus J3 originate from the reduction of MMP-9 expression in B16F10 cells.
RESUMO
We report the isolation and characterization of a novel knottin-type antimicrobial peptide from the yellow-spotted long-horned beetle Psacothea hilaris. A cDNA encoding a 56-mer knottin-type propeptide was identified and its predicted molecular mass and pI was 5.92 kDa and 8.28, respectively. A 34-mer mature peptide was also selected and named herein as psacotheasin. The antimicrobial activity of chemically synthesized psacotheasin against human bacterial pathogens was subsequently investigated. The results showed that psacotheasin exerted potent activities against both Gram-positive and Gram-negative bacterial strains. The present study suggests that psacotheasin can be applied to develop novel therapeutic agents.
Assuntos
Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Besouros/química , Sequência de Aminoácidos , Animais , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/farmacologia , Sequência de Bases , Motivos Nó de Cisteína , Perfilação da Expressão Gênica , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , RNA Mensageiro/química , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The caterpillar-shaped Chinese medicinal mushroom (DongChongXiaCao) looks like a worm in the winter and like a grass in the summer. The fruiting body has been regarded as popular folk or effective medicines used to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. The fruiting bodies of Paecilomyces tenuipes that formed on the living silkworm (Bombyx mori) host were used in this examination. This study was carried out to investigate the proximate composition, soluble sugar, amino acid and fatty acid profiles, and contents of the bioactive ingredient including adenosine and D-mannitol in the fruiting-bodies. The moisture content was 57.56%. Soluble sugars found were glycerol, glucose, mannitol and sucrose, and the contents exceeded 24 mgg(-1)dry weight. Total free amino acid content was 17.09 mg g(-1)dry weight. Arginine, glycine, proline and tyrosine were main amino acids. The content of oleic acid in fatty acids was high. Adenosine was more abundant in fruiting bodies than corpus.