ABSTRACT
<p><b>OBJECTIVE</b>To study the feasibility and the characteristics of recombinant baculovirus as spiral ganglion cells (SGC) gene transfer vector.</p><p><b>METHODS</b>After the generation of baculovirus- green fluorescent protein( Bac-GFP) according to Bac-to-Bac baculovirus expression system, SGC were infected by Bac-GFP with different multiplicities of infection (MOI) and different concentrations of sodium butyrate. The transfection cell rate and mean fluorescence strength (MFS) were detected by fluorescence microscopy and flow cytometry. Toxicity effects of recombinant baculovirus vectors and sodium butyrate on SGC were determined by spectroscopic measurement of 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-diphenytetrazoliumromide (MTF).</p><p><b>RESULTS</b>Baculovirus was able to infect primary SGC cultures. The dose-response characteristics of Bac-GFP were determined on SGC, and the expression level could be up-regulated by sodium butyrate. Infection with Bac-GFP in the absence or presence of sodium butyrate (< or =10 mmol/L) was considered to be non-cytotoxic to primary SGC. GFP had been expressed in SGC at 6 h post-infection and the highest numbers of cells expressing GFP were observed at approximately 48 h post-infection.</p><p><b>CONCLUSIONS</b>Baculovirus is a novel and promising tool for gene transferring into the cochlear nervous system both for studies of the function of foreign genes and the development of gene therapy strategies.</p>
Subject(s)
Animals , Rats , Baculoviridae , Genetics , Cells, Cultured , Gene Transfer Techniques , Genetic Vectors , Green Fluorescent Proteins , Genetics , Rats, Sprague-Dawley , Spiral Ganglion , Cell Biology , TransfectionABSTRACT
Objective To observe the effect of 5-aza-2'-deoxycytidine(5-aza-CdR)on the normal epithelial specific-1 gene(NES1)and the growth of human gastric cancer xenografts in nude mice,and to explore the possible anti-tumor mechanisms and search for new treatment for gastric cancer.Methods Human gastric caner xenograft model in nude mice was established and treated with 5-aza-CdR.The growth of xenografts in nude mice was observed,and the status of methylation and protein expression of NES1 gene were detected by MSP and immunohistochemistry respectirely.Results After treatment with 5-aza-CdR,the growth of the xenografts in nude mice was greatly inhibited(P