ABSTRACT
Objective: microRNA targeted to chronic myeloid leukemia Bcr-Abl oncogene were screened using the deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA). Methods: The proliferation inhibition effect of SAHA on chronic myeloid leukemia K562 cell was detected by MTS method, and the optimal concentration of SAHA reaction was determined. Western blot was used to detect the level of PARP protein, and making sure whether SAHA induced apoptosis of K562 cell. Effect of SAHA on Bcr-Abl Gene Transcription in K562 Cells was determined by Fluorescence Quantitative PCR. The online software Target Scan and real-time fluorescence quantitative PCR was used to screen Bcr-Abl-targeted microRNA. The viability of K562 cells and Bcr-Abl transcription levels were detected by MTS method and quantitative PCR respectively after selected microRNA were transfected into K562 cell. Results: SAHA significantly inhibited the proliferation of K562 cells and induced apoptosis, meanwhile SAHA significantly down-regulated the transcriptional level of Bcr-Abl gene. After treatment of K562 cells with SAHA, two microRNA, miR-192 and miR-6816, which could target Bcr-Abl, were screened by Target Scan and quantitative PCR. Additionally, SAHA induced the miRNAs to up-regulate 14.5 and 5.2 times, respectively. Transfection of miR-192 and miR-6816 to K562 cells significantly inhibited K562 cell viability and down-regulated the transcriptional level of Bcr-Abl gene. Conclusion: Acetylation inhibitor SAHA promoted the expression of miR-192 and miR-6816 in K562 cells by acetylation regulation, miR-192 and miR-6816 further down-regulated the transcription of Bcr-Abl gene, thereby inhibiting K562 cell proliferation and induced apoptosis.
