MiR-205-3p、miR-205-5p对头颈鳞状细胞癌细胞增殖、迁移及侵袭的影响

Abstract

Objective To investigate the effects of microRNA-205-3p (miR-205-3p) and microRNA-205-5p (miR-205-5p) on the cell proliferation, migration and invasion of head and neck squamous cell carcinoma (HNSCC). Methods The miRNA expression data were obtained from The Cancer Genome Atlas, and were used to determine miR-205 expression in HNSCC and paracancerous tissues. Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-205-3p and miR-205-5p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx. MiR-205-3p inhibitor, miR-205-5p inhibitor and miR-NC were transfected into Tu686 cells, and qRT-PCR was employed to evaluate the silence efficiency. In the following study, the cells were divided into four groups: miR-205-3p inhibitor and control, miR-205-5p inhibitor and control. Then the cell counting kit (CCK-8) assay, Transwell migration and invasion assays were carried out to examine the proliferation, migration and invasion abilities of the cells in the four groups. Results Compared with paracancerous tissues, the higher expression of miR-205 in HNSCC tissues was observed [M (QR): 61 012 (51 448) vs. 28 579 (35 959), Z=-6.420, P<0.001)]. The expression levels of miR-205-3p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.36±0.07, 0.20±0.06, 0.15±0.04, 0.25±0.04 and 1.00±0.00 respectively, with a significant difference (F=162.71, P<0.001). The expression levels of miR-205-5p in cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.20±0.01, 0.21±0.01, 1.06±0.18, 23.61±2.07 and 1.00±0.00 respectively, with a significant difference (F=371.81, P<0.001). Compared with 5-8F, 6-10B, CNE2 cells, the expression of miR-205-3p in Tu686 cells was higher than those in 6-10B, CNE2 cells (P=0.195; P=0.020), and lower than that in 5-8F cells (P=0.023), and the expression of miR-205-5p in Tu686 cells was the highest (P<0.001; P<0.001; P<0.001). The expressions of miR-205-3p and miR-205-5p in Tu686 cells were effectively downregulated by inhibitors, and the silence efficiencies were 87% and 83%, respectively. The absorbance (A) values of miR-205-3p inhibitor group on the first, second, third, fourth and fifth day were 0.26±0.06, 0.55±0.11, 1.52±0.13, 1.91±0.07, 2.14±0.24, and those of miR-NC group were 0.29±0.07, 0.78±0.11, 1.59±0.15, 1.95±0.08, 2.02±0.12. There were no significant differences between the two groups (t=0.506, P=0.639; t=2.459, P=0.070; t=0.573, P=0.597; t=0.655, P=0.548; t=-0.759, P=0.490). The cell proliferations of miR-205-5p inhibitor group on the first, second, third, fourth and fifth day were 0.30±0.08, 0.61±0.08, 0.85±0.08, 1.08±0.12, 1.16±0.18, and those of miR-NC group were 0.41±0.10, 0.78±0.14, 1.33±0.28, 1.87±0.09, 2.08±0.19. The cell proliferations of miR-205-5p inhibitor group on the third, fourth and fifth day were significantly lower than those of miR-NC group (t=3.665, P=0.017; t=12.223, P<0.001; t=7.825, P<0.001). Transwell migration assay demonstrated that downregulation of miR-205-3p had no significant effect on migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 192.00±28.49 and 188.40±22.52, respectively. There was no significant difference between the two groups (t=-0.160, P=0.877). Downregulation of miR-205-5p significantly decreased the migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 109.40±27.63 and 183.60±31.63, respectively, and the difference was statistically significant (t=3.951, P=0.004). Transwell invasion assay showed that the numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 93.40±10.24 and 96.20±16.56, respectively. There was no significant difference between the two groups (t=0.322, P=0.756). The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 53.00±17.80 and 94.40±14.38, respectively, and the difference was statistically significant (t=4.045, P=0.004). Conclusion Downregulation of miR-205-5p can inhibit the proliferation, migration and invasion abilities of Tu686 cells. However, downregulation of miR-205-3p has no significant effect on the proliferation, migration and invasion of Tu686 cells. Key words: Head and neck neoplasms; MicroRNAs; Cell proliferation; Cell movement; Neoplasm invasiveness