RESUMO
This study was performed to elucidate the effect of a lipid-soluble ginseng extract (LSGE) on cancer invasion and metastasis. The LSGE, even at noncytotoxic concentrations, potently inhibited invasion and migration of B16F10 mouse melanoma cells in a dose-dependent manner. In the presence of 3 µg/mL of LSGE, the invasion and migration of B16F10 cells were significantly inhibited by 98.1% and 71.4%, respectively. Furthermore, the LSGE decreased mRNA and protein levels of matrix metalloproteinase (MMP)-2 in B16F10 cells, leading to a decrease in MMP-2 activity. After B16F10 cells were intravenously injected in the tail vein of C57BL/6 mice, 1000 mg/kg/day of LSGE was orally administered for 13 days, after which lung metastasis of cancer cells was inhibited by 59.3%. These findings indicate that LSGE inhibits cancer cell invasion and migration in vitro and lung metastasis of melanoma cells in vivo by inhibiting MMP-2 expression.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Neoplasias Pulmonares/prevenção & controle , Metaloproteinase 2 da Matriz/metabolismo , Melanoma/tratamento farmacológico , Panax , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundário , Metaloproteinase 2 da Matriz/genética , Melanoma/genética , Melanoma/metabolismo , Melanoma/patologia , Camundongos Endogâmicos C57BL , Invasividade Neoplásica , Extratos Vegetais/farmacologia , RNA Mensageiro/metabolismo , SolubilidadeRESUMO
This study was performed to elucidate the anticancer mechanism of a lipid-soluble ginseng extract (LSGE) by analyzing induction of apoptosis and arrest of cell cycle progression using the NCI-H460 human lung cancer cell line. Proliferation of NCI-H460 cells was potently inhibited by LSGE in a dose-dependent manner. The cell cycle arrest at the G0/G1 phase in NCI-H460 cells was induced by LSGE. The percentage of G0/G1 phase cells significantly increased, while that of S phase cells decreased after treatment with LSGE. The expression levels of cyclin-dependent kinase2 (CDK2), CDK4, CDK6, cyclin D3 and cyclin E related to G0/G1 cells progression were also altered by LSGE. In addition, LSGE-induced cell death occurred through apoptosis, which was accompanied by increasing the activity of caspases including caspase-8, caspase-9 and caspase-3. Consistent with enhancement of caspase activity, LSGE increased protein levels of cleaved caspase-3, caspase-8, caspase-9, and poly-ADP-ribose polymerase (PARP). These apoptotic effects of LSGE were inhibited by the pan-caspase inhibitor Z-VAD-fmk. These findings indicate that LSGE inhibits NCI-H460 human lung cancer cell growth by cell cycle arrest at the G0/G1 phase and induction of caspase-mediated apoptosis.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Neoplasias Pulmonares/tratamento farmacológico , Panax , Extratos Vegetais/farmacologia , Clorometilcetonas de Aminoácidos/farmacologia , Antineoplásicos Fitogênicos/química , Apoptose/fisiologia , Caspase 3/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 8/efeitos dos fármacos , Caspase 8/metabolismo , Caspase 9/efeitos dos fármacos , Caspase 9/metabolismo , Inibidores de Caspase , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Inibidores de Cisteína Proteinase/farmacologia , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Fase G1/efeitos dos fármacos , Humanos , Lipídeos/química , Neoplasias Pulmonares/patologia , Extratos Vegetais/química , Poli(ADP-Ribose) Polimerases/efeitos dos fármacos , Poli(ADP-Ribose) Polimerases/metabolismo , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Fase S/efeitos dos fármacos , SolubilidadeRESUMO
Hemoglobin was hydrolyzed with Esperase and Flavourzyme as the endopeptidase and exopeptidase, respectively. The solubility of the heme-iron enriched peptide fraction decreased as the degree of hydrolysis of the hydrolysate increased. When the pH of a hydrolysate was adjusted to 5.0 after simultaneous hydrolysis with the two enzymes, the solubility of heme-iron enriched peptide was nearly zero, and 98% of the heme-iron enriched peptide fraction was recovered as a precipitate. These results indicated that an effective separation method for the production of heme-iron enriched peptide could be established by pH adjustment of the hemoglobin hydrolysate with high degree of hydrolysis.
Assuntos
Heme/química , Hemeproteínas/química , Hemoglobinas/química , Endopeptidases/química , Precipitação Fracionada , Concentração de Íons de Hidrogênio , Hidrólise , Serina Endopeptidases/química , Solubilidade , Fatores de TempoRESUMO
An efficient production method of heme-iron-enriched peptide was developed based on enzymatic hydrolysis. Hemoglobin hydrolysis, carried out stepwise with commercially available exopeptidase and endopeptidase, resulted in an increased degree of hydrolysis (DH). Exopeptidase-catalyzed protein hydrolysis formed low molecular weight peptides and amino acids. Different process parameters including dialysis and ultra- and diafiltration were evaluated. Heme/peptide ratio increased as molecular weight cut-off (MWCO) of the dialysis membrane increased. When the hydrolysate was dialyzed against sodium phosphate buffer, a higher heme/ peptide ratio was obtained. The heme/peptide ratio of the hydrolysate reached up to 25.4% when the dialysis was carried out with a membrane of 12-14 kDa MWCO. Also, the ratio was improved by the use of ultrafiltration and diafiltration on the pilot-scale.