Yang Yan,Zhang Dong,Gong Mingfu,Yang Hua,Zhang Song,Zou Liguang.Detection of transcriptional activity of hTERT promoter with MR imaging[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(04):364-368.
Yang Yan,Zhang Dong,Gong Mingfu,Yang Hua,Zhang Song,Zou Liguang.Detection of transcriptional activity of hTERT promoter with MR imaging[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(04):364-368.DOI:
Detection of transcriptional activity of hTERT promoter with MR imaging
Objective To explore the possibility of detecting the transcriptional activity of human telomerase reverse transcriptase (hTERT) promoter with MR imaging (MRI). Methods The Lenti-hTERT-Fth-flag-Puro vector was constructed and transfected into telomerase-positive human lung carcinoma A549 cells and telomerase-negative primary human periodontal ligament fibroblasts (HPDLFs)
with Lenti-CMV-Fth-flag-Puro vector used as control. Immunofluorescent staining and Western blotting were used to detect Fth-flag protein expression. Prussian blue iron staining was used for assaying iron accumulation. MRI was performed to determine the changes of T
*
2
WI signal intensity and T
*
2
value of cells in vitro. Results After transfected with Lenti-hTERT-Fth-flag-Puro vector
Fth-flag protein overexpression and iron accumulation increase were observed in transfected A549 cells but not in untransfected cells. MRI showed that the T
*
2
WI signal intensity and T
*
2
value of transfected A549 cells (SI: 1 340.86±49.21
T
*
2
value: 12.10±0.92) were significantly decreased as compared to untransfected A549 cells (SI: 2 049.08±87.58
T
*
2
value: 21.44±1.15) (
P
0.05). However
Fth-flag protein overexpression and iron accumulation increase were not observed in HPDLFs cells after transfected with Lenti-hTERT-Fth-flag-Puro vector. Accordingly
no significant difference of the T
*
2
WI signal intensity and T
*
2
value were observed between transfected HPDLFs cells (SI: 2 225.42±73.43
T
*
2
value: 24.01±1.13) and untransfected HPDLFs cells (SI: 2 148.48±54.74
T
*
2
value: 23.39±1.72) (P0.05). While transfected with Lenti-CMV-Fth-flag-Puro vector
A549 and HPDLFs cells both showed Fth-flag protein overexpression an
d iron accumulation increase
and the T
*
2
WI signal intensity and T
*
2
value of transfected cells (A549: 1 137.38±60.59
9.61±1.17; HPDLFs: 1 299.16±53.42
11.54±0.74) were both significantly decreased as compared to untransfected ones (
P
0.05). Conclusion It is achievable to detect transcriptional activity of hTERT promoter using MRI technology. The reporter gene imaging system used in this study may become a new prognostic method for cancer.