RESUMO
The effect of hyperthermic carbon dioxide (CO2) pneumoperitoneum in combination with 5-fluorouracil (5-FU) on the proliferation and invasion of colon cancer was explored. Colon cancer cell line SW-480 was sealed into the urine collection bag to simulate pneumoperitoneum with 100% CO2 under a pressure of 12 mmHg. The cells were divided into group A, CO2 at 37ËC; group B, CO2 at 43ËC; group C, 5-FU; group D, CO2 at 37ËC+5-FU; group E, CO2 at 43ËC+5-FU; and control groups under normal culture conditions. The cell proliferation was assessed by CCK-8 test; the cell apoptosis was tested by FACS analysis; the cell invasion was examined by Transwell assay; the expression of HSP-70, caspase-3, HIF-1α and MMP-9 proteins and genes were detected by western blot analysis and RT-PCR. The SW-480 cells were injected into nude mouse cecum subserosal to establish a colon cancer model. We applied 43ËC CO2 pneumoperitoneum or 5-FU intraperitoneal chemotherapy to intervene, detected the transplantation tumor growth and metastasis. The cell proliferation was inhibited in groups B, C, D and E, apoptosis was induced in groups B, C, D and E, the Transwell cell number decreased in groups B, C, D and E, the transplantation tumor weight and metastasis rate were inhibited in groups B, C, D and E, but all not in group A. The most significant change was observed in group E. Hyperthermic CO2 pneumoperitoneum was able to reinforce the inhibition of 5-FU on proliferation and invasion of colon cancer.
Assuntos
Dióxido de Carbono/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/terapia , Fluoruracila/uso terapêutico , Hipertermia Induzida/métodos , Pneumoperitônio Artificial/métodos , Animais , Linhagem Celular Tumoral , Neoplasias do Colo/patologia , Terapia Combinada , Humanos , Injeções Intraperitoneais , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
AIM: To prepare monoclonal antibody (mAb) against alpha-zearalanol (alpha-ZER) and develop an immunoassay for the detection of alpha-ZER and its analogues residues in food derived from animal tissues. METHODS: alpha-ZER was conjugated to BSA as immunogen to immunize BALB/c mice and mAb were prepared by hybridoma technique. mAb's characteristics (titer, Ig subclass, specificity and relative affinity) were identified by ELISA. Standard inhibitive cure was made and sensitivity of the mAb was identified. 37 samples derived from animal liver were detected for alpha-ZER residues by competitive ELISA established in the study. RESULTS: 8 hybridoma cell lines stably secreting anti-alpha-ZER mAb were obtained. The titer of one of them (4E5) was 5.142x10(7). The Ig subclass was IgG1. The mAb was specific for alpha-ZER and its analogues and had no cross-reactivity with other compounds.8 positive results were found from the 37 samples derived from animal liver which were negative detected by HPLC. CONCLUSION: Anti-alpha-ZER mAb has been prepared successfully. A rapid method using the mAb for detecting alpha-ZER and its analogues residues in animal tissues has been established.
Assuntos
Anticorpos Monoclonais/imunologia , Zeranol/análogos & derivados , Animais , Anticorpos Monoclonais/biossíntese , Especificidade de Anticorpos , Bovinos , Galinhas , Ensaio de Imunoadsorção Enzimática , Hibridomas/imunologia , Imunoglobulina G/biossíntese , Imunoglobulina G/imunologia , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Zeranol/química , Zeranol/imunologiaRESUMO
Angiopoietin-related growth factor (AGF) is a newly identified member of angiopoietin-related proteins (ARPs)/angiopoietin-like proteins (Angptls). AGF has been considered as a novel growth factor in accelerating cutaneous wound healing, as it is capable of stimulating keratinocytes proliferation as well as angiogenesis. But in our paper, we demonstrate that AGF stimulates keratinocytes proliferation only at high protein concentration, however, it can potently promote adhesion, spreading, and migration of keratinocytes, fibroblasts, and endothelial cells. Furthermore, we confirm that the adhesion and migration cellular events are mediated by RGD-binding integrins, most possibly the alpha(v)-containing integrins, by in vitro inhibition assays using synthetic competitive peptides. Our results strongly suggest that AGF is an integrin ligand as well as a mitogenic growth factor and theoretically participates in cutaneous wound healing in a more complex mechanism.