miR-17-5p promotes proliferation and migration of CAL-27 human tongue squamous cell carcinoma cells involved in autophagy inhibition under hypoxia

miR-17-5p促进CAL-27人舌鳞状细胞癌细胞增殖和迁移与缺氧条件下自噬抑制有关

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作者:Fawei Pang, Chao Liu, Yanjun Cui, Kun Gong, Guangping Liu, Yuanyuan Bian, Xiaoli Gao, Dongsheng Zhang

Abstract

Autophagy contributes to head and neck squamous cell carcinoma (HNSCC) development and progression. MiR-17-5p down-regulates Beclin-1 and thus plays an important role in autophagy, but little is known about the function and regulation of miR-17-5p in HNSCC autophagy. This study aimed to investigate the role of miR-17-5p on proliferation, migration, and autophagy under hypoxia in CAL-27 human tongue squamous cell carcinoma cells. CAL-27 cells were transfected with 50 nmol miR-17-5p mimics to overexpress miR-17-5p. Cell proliferation and migration were determined by CCK-8 and wound healing assays, respectively, under hypoxia. Autophagy induced by hypoxia was detected by transmission electron microscope and Beclin-1 mRNA and protein expressions. The miR-17-5p mimics successfully increased the expression of miR-17-5p in CAL-27 cells by almost 700 fold compared with the miRNA mimic negative control. After 3 days, cells transfected with the miR-17-5p mimics showed higher proliferation compared with controls (P < 0.05) under hypoxia. MiR-17-5p transfected CAL-27 cells had a higher migratory capacity compared with the control cells (P < 0.05) under hypoxia. Furthermore, transmission electron microscopy showed that miR-17-5p overexpression inhibited the formation of autophagosomes in hypoxic cells. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot showed that miR-17-5p overexpression inhibited the mRNA and protein expression of Beclin-1 in CAL-27 cells submitted to hypoxia. MiR-17-5p overexpression promoted the proliferation and migration of the CAL-27 cells, but inhibited autophagy under hypoxia.

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