YAP gene silencing to improve the drug sensitivity of imatinib resistant myeloid leukemia cells K562

doi:10.12092/j.issn.1009-2501.2018.09.007

Abstract

Abstract:

AIM: To explore the effect and mechanism of enhanced imatinib resistant K562 cells (K563/IMA) on imatinib sensitivity after YAP gene silencing, and to support the treatment of imatinib resistant myeloid leukemia. METHODS: Transfection of YAP gene silencing with siRNA-YAP was conducted by small interference RNA technique (siRNA) in K562/IMA. The control group (control), the siRNA negative control group (siRNA-NC) and the YAP siRNA group (siRNA-YAP) were established. The expression level of YAP protein and mRNA in each cell after silencing was identified by RT-QPCR and Western blot. The sensitivity of each cell to imatinib was detected by CCK-8, and the median inhibitory concentration of IC50 was calculated. Flow cytometry was used to detect the apoptosis rate of each group. The expression level of YAP, Bcl-2, Bax, cleaved-caspase3, caspase-3 and Cyto-C were detected by Western blot. The metastasis and invasion were detected by Transwell compartment. RESULTS: After the YAP gene silenced, the expression level of YAP protein and mRNA in the siRNA-YAP was significantly lower than that in the control and the siRNA-NC. The IC50 value of siRNA-YAP was significantly lower than that of control and siRNA-NC, which was statistically significant (P<0.05). The results of flow cytometry showed that the apoptotic rate of siRNA-YAP was significantly higher than that of control and siRNA-NC at the same dosage, which was statistically significance (P<0.05). The migration and invasiveness of K562/IMA cells in YAP silenced were decreased significantly. After imatinib intervene, the level of Bcl-2/Bax in siRNA-YAP cells was down-regulated, and the levels of cleaved-caspase3, caspase-3 and Cyto-C were up-regulated. Compared with control and siRNA-NC, it was statistically significant (P<0.05). CONCLUSION: To silence the YAP gene in imatinib resistant K562/IMA cells can restore the sensitivity of tumor cells to imatinib, and promote the activation of mitochondrial apoptotic pathway, resulting in the apoptosis of drug-resistant tumor cells.

Key words: YAP gene, imatinib, human myeloid leukemia cells, sensitivity

CLC Number:

WANG Jin, YANG Yi, GUAN Qiaobin, CHEN Qixu, GUO Li, HAN Chenyang. YAP gene silencing to improve the drug sensitivity of imatinib resistant myeloid leukemia cells K562[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(9): 1008-1014.

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