RESUMO
Infection efficiency is the key issue for gene delivery using adenovirus vector and usually unsatisfactory. In this study, recombinant adenoviruses encoding recombinant human EPO were prepared using the Adeasy system, and injected into the mammary gland of goats via the teat canal. BAPTA was used to treat the mammary gland to facilitate adenoviruses infection compared with EGTA. Milk serum was collected from the infected mammary gland and characterized by ELISAs and Western blotting. Expression level of rhEPO from the group treated by BAPTA was higher than that treated by EGTA.
Assuntos
Infecções por Adenoviridae/metabolismo , Adenoviridae/genética , Ácido Egtázico/análogos & derivados , Eritropoetina/genética , Cabras/metabolismo , Glândulas Mamárias Animais/efeitos dos fármacos , Leite/metabolismo , Animais , Western Blotting , Quelantes/farmacologia , Ácido Egtázico/farmacologia , Eritropoetina/metabolismo , Feminino , Vetores Genéticos/genética , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Animais/virologia , Distribuição Aleatória , Proteínas RecombinantesRESUMO
The research investigated the effect of Patrinia heterophylla Bunge (Valerianaceae) polysaccharides (PHB-P1) on U14-bearing mice. The tumor weight of mice treated with PHB-P1 (30, 60 mg/kg body weight) was significantly lower than that of the control group, a decrease of serum lactate dehydrogenase (LDH) activity was observed, and the serum alkaline phosphatase (AKP) level was increased slightly. The number of apoptotic tumor cells was significantly increased in the mice by treatment of PHB-P1 (30, 60 mg/kgbw). Cell cycle analysis showed the accumulation of tumor cells in the G2/M phase and a relative decrease of the S phase. By the immunohistochemical analysis, PHB-P1 (30, 60 mg/kgbw) might up-regulate the expression of p53 and Bax, and significantly inhibited the expression of Bcl-2 in tumor tissues. In conclusion, PHB-P1 could inhibit tumor growth and induce tumor cell apoptosis.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Carcinoma/tratamento farmacológico , Patrinia/química , Fitoterapia , Extratos Vegetais/uso terapêutico , Polissacarídeos/uso terapêutico , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma/sangue , Carcinoma/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Camundongos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Distribuição Aleatória , Carga Tumoral/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos dos fármacos , Neoplasias do Colo do Útero/sangue , Neoplasias do Colo do Útero/patologia , Proteína X Associada a bcl-2/metabolismoRESUMO
We have examined the effects of the crude polysaccharides isolated from Solanum nigrum Linne (SNL-P) in vitro and in vivo against U14 cervical cancer. SNL-P showed no antiproliferative effects in vitro at a dose up to 1 mg/ml. In vivo administration with SNL-P (90, 180, 360 mg/kg b.w., p.o.) decreased the number of ascites tumor cells and prolonged the survival time of U14 cervical-cancer-bearing mice. FACScan flow cytometer analysis showed that most of the ascites tumor cells were arrested in G2/M phase of cell cycle and the ratio of CD4+/CD8+ peripheral blood T-lymphocyte subpopulations were restored following treatment of SNL-P. Furthermore, the treatment with SNL-P also caused a significant increment in IFN-gamma (p < 0.01, 90, 180 and 360 mg/kg b.w.) and a remarkable decrease in Il-4 (p < 0.01, 90, 180 mg/kg b.w.; p < 0.05, 360 mg/kg b.w.) by the method of ELISA. These data showed that SNL-P possess potent antitumor activity and SNL-P might exert antitumor activity via activation of different immune responses in the host rather than by directly attacking cancer cells on the U14 cervical cancer bearing mice. Thus, SNL-P could be used as an immunomodulator and an anticancer agent.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Solanum nigrum , Animais , Antineoplásicos Fitogênicos/imunologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Interferon gama/sangue , Interleucina-4/sangue , Camundongos , Extratos Vegetais/imunologia , Polissacarídeos/imunologia , Linfócitos T/efeitos dos fármacos , Neoplasias do Colo do Útero/tratamento farmacológicoRESUMO
The aim of our study was to investigate the effect of Patrinia scabra Bunge polysaccharide (PSB-P2) on cervical cancer cell (U14)-bearing mice. The tumor weight of mice treated with PSB-P2 (40, 80 mg/kg b.w.) was significantly lower than that of the control group and serum lactate dehydrogenase (LDH) activity was decreased, while serum alkaline phosphatase (AKP) level was only changed slightly. Meanwhile, the number of apoptotic tumor cells was significantly increased in the mice by the treatment of PSB-P2 (40, 80 mg/kg b.w.). At the same time, cell cycle analysis showed the accumulation of tumor cells in the G0/G1 phase and a relative decrease in the S phase. On the other hand, using the reverse transcription-polymerase chain reaction (RT-PCR) assay, PSB-P2 (40, 80 mg/kg b.w.) showed the up-regulation of p53 and Bax, and significant inhibition of Bcl-2 in tumor tissues. It suggests a possible mechanism of the inhibitory effect of PSB-P2 on tumor growth.