Volume 14, Issue 6 (February-March 2012)
J Arak Uni Med Sci 2012, 14(6): 104-116 |
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Moosavi M A, Seyed Gogani N, Asvadi Kermani I, Asadi M. Nucleostemin gene silencing by siRNA and growth inhibition, cell cycle arrest, and apoptosis induction of K562 leukemia cell line. J Arak Uni Med Sci 2012; 14 (6) :104-116
URL: http://jams.arakmu.ac.ir/article-1-905-en.html
URL: http://jams.arakmu.ac.ir/article-1-905-en.html
1- , moosav_m@tabrizu.ac.ir
2- Hematology and Oncology Research Center, The Medical Science University of Tabriz,
3- Department of Pharmacology, The Medical Science University of Tabriz,
2- Hematology and Oncology Research Center, The Medical Science University of Tabriz,
3- Department of Pharmacology, The Medical Science University of Tabriz,
Abstract: (14448 Views)
Background: Nucleostemin plays a critical role in controlling proliferation and self-renewal of stem cells and cancer cells. Thus, inhibition of nucleostemin expression could be a potent therapeutic approach in cancer treatment. In the present study, the effects of nucleostemin gene silencing in K562 cell line were studied.
Materials and Methods: In this experimental study, after transfecting NS-specific siRNA into K562 cells, changes in nucleostemin gene expression pattern were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Trypan blue exclusion test, MTT assay, and fluorescent microscopy were used to evaluate the growth inhibition and apoptosis of K562 cells, respectively. Flow-cytometery was utilized for evaluating the effects of nucleostemin gene silencing on cell cycle.
Results: The results showed the high expression of nucleostemin gene in K562 cells. NS-siRNA transfection into K562 cells at 200 nM inhibited the nucleostemin mRNA level up to 55% after 48 hours when compared to corresponding control cells. Forty eight hours after transfection, the cell growth decreased up to 33.7%. In addition, the silencing of nucleostemin induced G1 cell cycle arrest. Furthermore, fluorescent microscopy assays indicated that apoptosis occurred 48 hours after silencing nucleostemin gene expression.
Conclusion: Noticing the potent growth inhibitory and apoptotic effects of nucleostemin siRNA in human myeloid leukemia K562 cells, silencing this gene can be a potential target for inhibiting K562 cells as the stem cell model of chronic myeloid leukemia.
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