RESUMO
GalNAc-transferase-7 (GALNT7) is essential for the regulation of cell proliferation and has been implicated in tumorigenesis. However, the role of GALNT7 in the development and progression of nasopharyngeal carcinoma (NPC) remains unclear. Our previous study showed that GALNT7 was a putative target of miR-494, which was confirmed by luciferase reporter assay. In the present study, we demonstrated that in vitro knockdown of GALNT7 significantly inhibited the proliferation, colony formation, migration, and invasion of NPC-derived cells. In vivo tumorigenicity assay showed that miR-494 and GALNT7-small interfering RNA (siRNA) reduced tumor growth in nude mice. Taken together, our results provided new evidence for an oncogenic role of GALNT7 in NPC.
Assuntos
Carcinogênese/genética , MicroRNAs/genética , N-Acetilgalactosaminiltransferases/genética , Neoplasias Nasofaríngeas/genética , Animais , Apoptose , Carcinogênese/metabolismo , Carcinoma , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos Nus , N-Acetilgalactosaminiltransferases/metabolismo , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Transplante de Neoplasias , Interferência de RNA , Carga Tumoral , Polipeptídeo N-AcetilgalactosaminiltransferaseRESUMO
AIM: To investigate intraperitoneal transplantation of microencapsulated hepatic-like cells from human umbilical cord blood for treatment of hepatic failure in rats. METHODS: CD34+ cells in umbilical cord blood cells were isolated by magnetic cell sorting. In the in vitro experiment, sorted CD34+ cells were amplified and induced into hepatic-like cells by culturing with a combination of fibroblast growth factor 4 and hepatocyte growth factor. Cultures without growth factor addition served as controls. mRNA and protein levels for hepatic-like cells were analyzed by reverse transcription-polymerase chain reaction, immunohistochemistry and immunofluorescence. In the in vivo experiment, the hepatic-like cells were encapsulated and transplanted into the abdominal cavity of acute hepatic failure (AHF) rats at 48 h after D-galactosamine induction of acute hepatic failure. Transplantation with PBS and unencapsulated hepatic-like cells served as controls. The mortality rate, hepatic pathological changes and serum biochemical indexes were determined. The morphology and structure of microcapsules in the greater omentum were observed. RESULTS: Human albumin, alpha-fetoprotein and GATA-4 mRNA and albumin protein positive cells were found among cultured cells after 16 d. Albumin level in culture medium was significantly increased after culturing with growth factors in comparison with culturing without growth factor addition (P < 0.01). Compared with the unencapsulated group, the mortality rate of the encapsulated hepatic-like cell-transplanted group was significantly lower (P < 0.05). Serum biochemical parameters, alanine aminotransferase, aspartate aminotransferase and total bilirubin in the encapsulated group were significantly improvement compared with the PBS control group (P < 0.01). Pathological staining further supported these findings. At 1-2 wk post-transplantation, free microcapsules with a round clear structure and a smooth surface were observed in peritoneal lavage fluid, surviving cells inside microcapsules were found by trypan blue staining, but some fibrous tissue around microcapsules was also detected in the greater omentum of encapsulated group by hematoxylin and eosin staining. CONCLUSION: Transplantation of microencapsulated hepatic-like cells derived from umbilical cord blood cells could preliminarily alleviate the symptoms of AHF rats.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos/métodos , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Composição de Medicamentos/métodos , Hepatócitos/transplante , Falência Hepática Aguda/terapia , Animais , Antígenos CD34/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Sangue Fetal/citologia , Sangue Fetal/efeitos dos fármacos , Sangue Fetal/imunologia , Fator 4 de Crescimento de Fibroblastos/farmacologia , Galactosamina/efeitos adversos , Fator de Crescimento de Hepatócito/farmacologia , Humanos , Falência Hepática Aguda/induzido quimicamente , Modelos Animais , Ratos , Ratos Sprague-Dawley , Resultado do TratamentoRESUMO
OBJECTIVE: To establish an in vitro model for the development of mouse spermatogenic cells into sperm by using the immunodefective mouse as the incubator. METHODS: Tissue grafting was performed using testis from 1-2 days old Kun-ming mice as donor tissue and immunodefective mice as recipients; the expression of TESK1 mRNA in grafts was determined by RT-PCR and the spermatogenesis further observed with histological analysis of grafts. RESULTS: Molecular biological and histological analyses showed that grafts post-grafting not only expressed TESK1 mRNA as in normal mouse testis, but also exhibited similarities in the structure of seminiferous tubules and component of spermatogenic cells, including sperms. CONCLUSION: Spermatogenic cells heterotopically grafted in vitro could continuously grow and complete spermatogenesis and finally develop into sperm.