신경줄기세포(HB1.F3)에서 나트륨옥소 공동수송체 도입유전자 발현 (Expression of Sodium/iodide Symporter Transgene in Neural Stem Cells) |
Author |
김윤희, 이동수, 강주현, 이용진, 정준기, 이명철, |
Yun Hui Kim, B.S., Dong Soo Lee, M.D.Ph.,D., Joo Hyun Kang, Ph.D., Yong Jin Lee,M.S., June-Key Chung, M.D.,Ph.D., Myung Chul Lee, M.D.,Ph.D. |
Affiliation |
서울대학교 뇌과학 협동과정, 서울대학교 의과대학 핵의학교실 Program in Neuroscience, Seoul National University, Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea |
Abstract |
The ability to noninvasively track the migration of neural progenitor cells would have significant
clinical and research implications. We generated stably transfected F3 human neural progenitor cells with
human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns
of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation.
Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou
University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome
Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To
investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent
(5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was
measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was
higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS
gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to 20μM, up to 62.5nM,
respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These
results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell
passage. Therefore, the gene expression of exogenous gene transferred human stem cell might be affected
to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture
conditions. |
Keyword |
neural stem cell, human sodium/iodide symporter, promoter methylation, histone deacetylation |
Full text Article |
3801099.pdf
|
|
|