Effects of microRNA-204-3p and microRNA-205-5p on the cell proliferation, migration and invasion of head and neck squamous cell carcinoma / 国际肿瘤学杂志

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.

The expression of ubiquitin in laryngeal squamous cell carcinoma with lymph node metastasis and its clinical significance / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the ubiquitin expression in laryngeal squamous cell carcinoma (LSCC) whether along with local lymph node metastasis, and further study its correlation with local lymph node metastasis and other clinicopathological parameters in laryngeal squamous cell carcinoma.@*METHOD@#We detected the different expression level of ubiquitin in paraffin specimens between 19 cases of LSCC associated with cervical lymph node metastasis LSCC(N+) and 20 cases of LSCC not associated with cervical lymph node metastasis LSCC(N-) by immunohistochemical staining combined with stereology image analysis system. Statistics were analyzed by student test, variance analysis and ROC curve.@*RESULT@#Ubiquitin expression in LSCC(N+) was significantly higher than LSCC(N-) (P < 0.01); their expression level was not correlated with age,history of tobacco, alcohol addiction, clinical stage and primary site,etc.@*CONCLUSION@#Ubiquitin was significantly up-expressed in LSCC(N+) than ILSCC (N-), which may imply that it is one of the important elements in mechanism of lymph node metastasis in LSCC.

The expression and correlation of HMGB1 and VEGF protein in laryngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression and biological significance of HMGB1 and VEGF protein in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and further study the correlation between HMGB1 and VEGF protein.@*METHOD@#The expression of HMGB1 and VEGF protein was evaluated by immunohistochemical staining in 69 cases of LSCC specimens and 15 cases of adjacent epithelial tissue samples, and futher correlated with clinicopathologic parameters.@*RESULT@#The positive rates of HMGB1 and VEGF in LSCC tissues were significantly higher than those in adjacent non-cancerous mucosa (P 0. 05). There was a positive correlation between the expression of HMGB1 and VEGF (P < 0.05). The Kaplan-Meier survival analysis showed that patients with strong expression of HMGB1 or VEGF had poorer overall survival compared with that in patients with relative low HMGB1 or VEGF expression (P < 0.05). Multivariate COX regression analysis revealed that both lymph node metastasis and HMGB1 expression were independent prognostic factors for patients with LSCC.@*CONCLUSION@#This study demonstrated that HMGB1 and VEGF protein overexpression were closely associated with clinical stage, metastasis and poorer prognosis in patients with LSCC. Increased expression of these two proteins in LSCC suggested that HMGB1 and VEGF might play a critical role in the initiation and progression of LSCC.