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The Effect of Recombinant Adenovirus AdE-SH2-Caspase8-HA-GFP on the Proliferation of Imatinib-resistant K562/G01 Cell Line
Wang Lin, Luo Qiuping, Huang Zhenglan, Li Hui, Liu Zhanglin, Zu Bailin, Feng Wenli*, Xiang Hua*
Key Laboratory of Medical Diagnostics of Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University,Chongqing 400016, China
Abstract: To study the effect of SH2-Caspase8 fusion protein expressed by recombinant adenovirus dE-SH2-Caspase8-HA-GFP on the proliferation of K562/G01 cell line， which is a BCR/ABL positive chronic myeloid leukemia cell line and resistant to imatinib. Infection efficiency was observed under fluorescent microscopy. The expression of fusion protein was analyzed by Western blot. Cell growth was detected by MTT test and cell counts. Cell cycles were determined by flow cytometry. The ability of cell colony was assessed by colony-forming assay. As a result， the infection efficiency of AdE-SH2-Caspase8-HA-GFP on K562/G01 cells was high as confirmed by fluorescent microscopy. SH2-Caspase8 fusion protein was expressed correctly in K562/G01 cells. As compared to control groups， AdE-SH2-Caspase8-HA-GFP adenovirus significantly inhibited cell growth. The result of FCM showed that cells in G1 phase increased， while cells in S phase and G2 phase decreased. AdE-SH2-Caspase8-HA-GFP adenovirus inhibited the colony-forming ability of K562/G01 cells obviously. Together all， AdE-SH2-Caspase8-HA-GFP which expresses SH2-Caspase8 fusion protein can significantlyinhibit the proliferation of K562/G01 cells.
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