摘要
目的:以复制缺陷型重组腺病毒为载体,使用人野生型p53基因、人GM-CSF基因和B7-1基因为目的基因,探讨三基因在胰腺癌细胞BxPC-3中的表达。方法:免疫组织化学方法检测外源性p53蛋白在BxPC-3细胞中表达。ELISA方法测定GM-CSF表达量;流式细胞术(flow cytometry,FCM)检测BxPC-3细胞转染B7-1的表达效果。结果:重组腺病毒介导的p53基因、GM-CSF基因、B7-1基因在BxPC-3细胞中得到高效表达。结论:重组腺病毒介导的外源基因在BxPC-3细胞中得到高效的表达,为进一步探讨其体内外治疗胰腺癌的实验研究提供了理论基础。
Objective: To study the expression of human wild-type p53,GM-CSF and B7-1 genes mediatedly adenovirus vetor on human pancreatic carcinoma cell line BxPC-3.Methods: The expression of p53 protein was examined by immunohistochemical analysis.The expression of GM-CSF was detected by sandwich ELISA analyses with culture supernatant.The expression of B7-1 was detected with FCM. Results: BxPC-3 cells were transfected with DMEM,Ad-GFP,AD-WTp53-GMCSF-B7-1.The cells transfected with the Ad-WTp53-GMCSF-B7-1 resulted in positive expressing the wild-type p53 protein,GM-CSF and B7-1,but not in DMEM and Ad-GFP transfected cells. Conclusion: The expression on human pancreatic carcinoma with adenovirus-mediated human wild-type p53,GM-CSF and B7-1 genes was more effective.These results provided a basis for further ex vivo and in vivo studies of combined gene therapy for pancreatic carcinoma.
出处
《江苏大学学报(医学版)》
CAS
2009年第6期489-492,共4页
Journal of Jiangsu University:Medicine Edition
基金
江苏大学医学科技发展基金资助项目(JLY20050034)