中国肿瘤生物治疗杂志2009，Vol.16Issue(5)：452-457,6.DOI:10.3872/j.issn.1007-385X.2009.05.006
hTERT启动子调控的异种抗原基因α-1,3GT表达载体的构建及其在肺癌细胞中的靶向表达
Construction of xenoantigen α-1,3GT gene expression vector regulated by hTERT promoter and its targeting expression in human lung cancer cells
摘要
Abstract
Objective:To construct an xenoantigen synthetase α-1,3 galactosyltransferase (α-1,3GT) eukaryotic expression vector regulated by human telomerase catalytic subunit (hTERT) promoter, and to investigate its targeting expression of α-1,3GT in lung cancer cell lines. Methods: Previously prepared and confirmed pig α-1,3GT gene was inserted into pEGFP-hTERTp plasmid to construct eukaryotic expression vector pEGFP-hTERTp-GT. pEGFP-hTERTp-GT and pEGFP-N1-GT (α-1,3GT eukaryotic expression vector under the control of CMV promoter) were transfected into telomerase-positive human lung adenocarcinoma A549 cells and telomerase-negative human embryonic lung fibroblast MRC-5 cells. 1,3-GT mRNA expression in the transfected cells was detected by RT-PCR. Expression of α-gal antigen in transfected cells was examined by immunofluorescence and flow cytometry. Results: pEGFP-hTERTp-GT eukaryotic expression vector was successfully constructed. Both A549 and MRC-5 cells transfected with pEGFP-N1-GT showed expression of α-1,3GT mRNA; A549 cells but not telomerase-negative MRC-5 cells expressed α-1,3GT mRNA after transfection with pEGFP-hTERTp-GT. Furthermore, both A549 and MRC-5 cells transfected with pEGFP-N1-GT showed expression of xenoantigen α-gal; A549 but not MRC-5 cells expressed xenoantigen α-gal after transfection with pEGFP-hTERTp-GT (P<0.01). Conclusion: α-1,3GT gene under the regulation of hTERT promoter can be specifically expressed in telomerase-positive lung cancer cell lines, which can induce production of xenoantigen α-gal.关键词
人端粒酶逆转录酶/肺肿瘤/α-半乳糖基/α-1,3半乳糖基转移酶/靶向性表达Key words
human telomerase reverse transcriptase (hTERT)/ lung neoplasms/ α-galactosyl (α-gal)/ α-1,3 galactosyltransferase/ targeting expression分类
医药卫生引用本文复制引用
朱圣明,郑鸿,秦凤,王宇,王竹,骆志国,丁珺,王艳萍..hTERT启动子调控的异种抗原基因α-1,3GT表达载体的构建及其在肺癌细胞中的靶向表达[J].中国肿瘤生物治疗杂志,2009,16(5):452-457,6.基金项目
国家自然科学基金资助项目(No. 30270589,No. 30470762).Supported by the National Natural Science Foundation of China (No. 30270589, No. 30470762) （No. 30270589,No. 30470762）
