Study on transcriptional activities of tumor-specific hTERT and survivin promoter in human lung cancer A549 cell line
- VernacularTitle:肿瘤特异性hTERT启动子与Survivin启动子在肺癌A549细胞中的转录活性研究
- Author:
Yixuan LI
- Publication Type:Journal Article
- Keywords:
hTERT;
Survivin gene;
Promoter;
Lung cancer;
Gene therapy
- From:
Journal of Chongqing Medical University
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objectives:To compare the transcriptional activities of tumor-specific hTERT promoters and survivin promoter in human lung cancer A549 cells,in order to lay some groundwork for targeting gene therapy in human lung cancer.Methods:The 1 084 bp fragment of hTERT promoter and 980bp survivin promoter from A549 cells were acquired by PCR amplification.The eukaryotic expression plasmids(pGL-H-RED,pGL-LH-RED and pGL-S-RED)which have Red Fluorescin reporter gen(eRed)and are modulated by 263 bp hTERT promoter,1084bp hTERT promoter and 980bp survivin promoter respectively were constructed based on the pGL3-hTERT which has 263bp hTERT promoter from HepG2 cells.These recombinant plsmids were transfected into A549 and MRC-5 cells with liposome.After 72 hours,the expression level of red fluorescin target protein modulated by 3 promoters at the same time was analyzed by Imagepro-Plus6.0 software.Results:The red fluorescin target protein was observed in transfected A549 cells,but there was no red fluorescence in transfected MRC-5 control cells.The pGL-S-RED had the highest expression activity in A549 cells and the strength rate(IOD)of red fluorescence of A549 cells transfected by pGL-S-RED,pGL-LH-RED and pGL-H-RED were 201.17,171.70,and 136.34 respectively.Conclusion:Our data reveals that the cloned hTERT and survivin promoter are tumor-specific,and the survivin promoter has the highest transcriptional activities in lung cancer A549 cells,and may serve as a useful tool for transcriptional targeting gene therapy of human lung cancer.
