Effects of microRNA-204-3p and microRNA-205-5p on the cell proliferation, migration and invasion of head and neck squamous cell carcinoma
10.3760/cma.j.issn.1673-422X.2018.11.001
- VernacularTitle: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
- Author:
Changyun YU
1
;
Yong LIU
;
Hua CAO
Author Information
1. 450052,郑州大学第一附属医院耳鼻咽喉科
- Keywords:
Head and neck neoplasms;
MicroRNAs;
Cell proliferation;
Cell movement;
Neoplasm invasiveness
- From:
Journal of International Oncology
2018;45(11):641-646
- CountryChina
- Language:Chinese
-
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 downregnlation 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.
