RESUMO
Granzyme B and perforin, two major effector molecules in the granule-mediated cytolytic pathway, are thought to be involved in suppression of tumor progression. In this study, the pGEX-4T-1 expression vector was used to express full-length human perforin or granzyme B as a GST-tagged fusion protein in Escherichia coli (E. coli). GST-tagged proteins were induced with IPTG and purified by GSTrap 4B columns. Purified fusion proteins migrated at the predicted molecular mass on SDS-PAGE and were recognized by specific antibodies. Moreover, the fusion proteins can induce apoptosis and directly inhibit the growth of human laryngeal cancer Hep-2 cells in vitro. These results suggest that active perforin and granzyme B fusion proteins can be produced in E. coli and exhibit anticancer potential in laryngeal cancer cells.
Assuntos
Antineoplásicos/farmacologia , Granzimas/metabolismo , Perforina/metabolismo , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/farmacologia , Antineoplásicos/isolamento & purificação , Antineoplásicos/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Escherichia coli/genética , Glutationa Transferase , Granzimas/genética , Granzimas/isolamento & purificação , Humanos , Neoplasias Laríngeas , Perforina/genética , Perforina/isolamento & purificação , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismoRESUMO
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RESUMO
Granzyme B and perforin, two of the most important components, have shown anticancer properties in various cancers, but their effects in laryngeal cancer remain unexplored. Here we decided to examine the effects of Granzyme B and perforin in Hep-2 cells and clarify the role of perforin and granzyme B in the tumorigenicity of laryngeal cancer cell line. Hep-2 cells were transfected with pVAX1-PIG co-expression vector (comprising perforin and granzyme B genes), and then the growth and apoptosis of these Hep-2 cells were evaluated. The tumorigenicity of Hep-2 cell line co-expressing perforin and granzyme B genes was tested in BALB/c nu/nu mice. We found that the co-expression of perforin and granzyme B genes could obviously inhibit cell focus formation and induce cell apoptosis in Hep-2 cells. Furthermore, after subcutaneous injection of Hep-2 cells transfected with pVAX1-PIG, an extensive delay in tumor growth was observed in BALB/c-nu/nu mice. Moreover, our studies demonstrated that the anticancer activity of perforin and granzyme B was sustainable in vivo as tumor development by inducing cell apoptosis. Taken together, our data indicate that the co-expression of perforin and granzyme B genes exhibits anticancer potential, and hopefully provide potential therapeutic applications in laryngeal cancer.
Assuntos
Apoptose/genética , Carcinogênese/metabolismo , Granzimas/biossíntese , Neoplasias Laríngeas/metabolismo , Perforina/biossíntese , Animais , Carcinogênese/genética , Linhagem Celular Tumoral , Citometria de Fluxo , Granzimas/genética , Xenoenxertos , Humanos , Imuno-Histoquímica , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Microscopia Eletrônica de Transmissão , Perforina/genética , TransfecçãoRESUMO
A link tooth wheel-cylinder non-thermal plasma reactor was set up to investigate the degradation of phenol in the mists. In addition, the decomposition efficiency of phenol, TOC removal, and byproduct formation were investigated. The stable discharge was achieved in both air and the mist condition. The decomposition efficiency and TOC removal increased with increasing the input power. For the input power of 3.6 W, the phenol decomposition and TOC removal reached 90% and 47%, respectively. Phenol degradation byproducts were identified as small molecular organic acids, including formic acid, acetic acid, and oxalic acid. Their masses in the trapped solutions first increased and then decreased slightly with increasing the input power. Therefore, the biodegradation capacity of the phenol degradation byproducts can be improved.