Analysis the role of some effective microRNAs on the Hippo signaling pathway in Chronic Myeloid Leukemia K۵۶۲ cell line treated with NVP-AUY۹۲۲

Introduction Chronic Myeloid leukemia is a proliferative neoplasm caused by an aberrant expression of an oncogene called BCR-ABL that is formed after a chromosomal translocation. MicroRNAs (miRNAs) are a group of small non-coding RNAs that modulate the activity of specific mRNA target with important roles in a wide range of physiologic and pathological processes. They affect various signaling cascades during the progression of neoplasms by the regulation of gene expression at the post-transcriptional level. The Hippo signaling pathway plays important role in organ development, tissue regeneration, and human diseases such as cancer and consist of a highly conserved kinase cascade (STK۳/۴, MOB۱A/B, LATS۱/۲) and downstream transcription co-activators (YAP and TAZ). Heat shock protein ۹۰ (HSP۹۰) is a molecular chaperone with different client proteins. It helps these client proteins to fold correctly and maintain their appropriate conformation which is essential for various events such as cell cycle control, signal transduction and gene transcription. NVP-AUY۹۲۲ is a novel HSP۹۰ inhibitor active against tumor growth and metastasis. Here, we aim to report the role of expression level of three microRNAs (miR-۲۹b-۳p, miR-۵۹۰-۳p, miR-۱۸۲-۵p) and their potential target genes related to the Hippo signaling pathway in K۵۶۲ cell line treated with NVP-AUY۹۲۲. Methods K۵۶۲ cells were cultured and then treated with different concentration of NVP-AUY۹۲۲. IC۵۰ concentration of NVP-AUY۹۲۲ was calculated after ۴۸ hour by Trypan Blue exclusion dye. Expression level of microRNAs and their Hippo target genes were evaluated after ۴۸ hour treatment with NVP-AUY۹۲۲ in K۵۶۲ cell lines by Real-time PCR technique. U۶ and GAPDH were chosen as control genes and all the primers were designed with oligo ۷ software. Results ۵۰?ll viability was obtained at ۲۵۰nM concentration of NVP-AUY۹۲۲. miR-۲۹b-۳p and miR-۱۸۲-۵p were upregulated about ۲.۸ and ۲ times more after ۴۸hr treatment with NVP-AUY۹۲۲ compared to non-treated cells. In contrary miR-۵۹۰-۳p was downregulated about ۹۰?ter ۴۸hr treatment with NVP-AUY۹۲۲. Evaluation of expression of Hippo pathway components showed up regulation of MOB۱A/B, major targets of miR-۲۹b-۳p and miR-۱۸۲-۵p. Also, LATS۱/۲ and STK۳/۴ were up regulated as major targets of miR-۵۹۰-۳p. Conclusion It seems that microRNAs and their target genes can modulate cell proliferation in NVP-AUY۹۲۲ treated cells and it could be nominated as a therapeutic strategy for CML targeted treatment.

کلیدواژه ها:

Chronic Myeloid leukemia ، Hippo signaling pathway ، microRNA ، NVP-AUY۹۲۲