Identification of miRNA Targeting FTO-Downregulation and Regulating Biological Behavior of Breast Cancer Cells
LU Shuangxin, HUAI Ruiping, CHEN Mengyang, LI Gang, YANG Hui, XIONG Lili*
This study aimed to identify miRNA that targeted FTO and its role in biological behaviors of breast cancer cells. Bioinformatic analysis was used to screen out the survival-related RNA methylation regulator FTO. CCK-8 assay was performed to evaluate the proliferation of breast cancer cells with FTO-downregulation. miR-504-5p was subsequently predicted as a potential miRNA targeting FTO. qRT-PCR and Western blot were applied to investigate the effects of miR-504-5p on the expression of FTO. Dual-luciferase reporter assay was utilized to validate the relationship between miR-504-5p and FTO. CCK-8 assay, Transwell assay and flow cytometry were performed to evaluate the influence of miR-504-5p on the proliferation, migration, apoptosis and cell cycle distribution of breast cancer cells. The results indicated that FTO low-expressed breast cancer patients had more favorable survival rate than the FTO high-expressed breast cancer patients. miR-504-5p was able to down-regulate the expression of FTO mRNA or protein and it binded with FTO in the position of 5 927–5 933 of FTO 3ʹ-UTR. miR-504-5p attenuated the proliferation and migration, meanwhile promoted the apoptosis of breast cancer cells, blocked the cell cycles in G0/G1. In summary, this study found that miR-504-5p could down-regulate the expression of FTO, inhibit the proliferation and migration of breast cancer cells, promote the apoptosis of breast cancer cells, and block the cell cycle of breast cancer cells. This study may provide potential reference values for molecular mechanism study and therapy of breast cancer.
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