Flt3配体重组腺病毒感染胃癌细胞的体外研究

Flt3 Ligand Gene Infection of MFC in Vitro Mediated by Recombinant Adenovirus Vector

目的 研究Flt3配体重组腺病毒(AdmFL)体外感染前小鼠前胃癌细胞的情况。方法 体外分别以不同MOI(mutiplicity of infection)的AdmFL感染胃癌细胞,荧光显微镜下计算感染率,对体外感染后的胃癌细胞连续计数并绘制生长曲线,RT-PCR检测转染后的细胞内mRNA表达,ELISA法检测培养上清中mFl蛋白的浓度。结果 MOI为10、100、500的AdmFL感染小鼠胃癌细胞,荧光显微镜下2 d后根据GFP的表达计算的感染率分别是5％、60％和98％;以MOI为100的AdmFL对体外感染后的胃癌细胞无直接的生长抑制作用;在感染后的胃癌细胞内RT-PCR扩增到了mFL的mRNA,ELISA法检测到mFL蛋白的浓度为60.3±5.2 ng／ml。结论 Flt3配体重组腺病毒能有效地感染小鼠前胃癌细胞,并能介导所携带的mFL高效表达。

Objective To study Flt3 ligand gene infection of MFC in vitro mediated by recombinant adenovirus vector. Method In vitro infection of murine gastric carcinoma cell line MFC with AdmFL at MOI 10、100 and 500, respectively, measured by GFP expression. The mRNA was detected by RT-PCR. The level of mFL protein was check out by ELISA. Results In vitro Infection with AdmFL at MOI 10、100 and 500, was able to lead to 5%、60% and 95% infection rate respectively. There was no direct growth inhibition on MFC infected by AdmFL at MOI 100. The mFL protein was 60.3±5. 2 ng/ml in the infected MFC cell culture supernatant. Conclusion All data suggest AdmFL can infect MFC efficiently and mediate the expression of mFL in vitro.