Targeted-knockdown of Yes-associated protein inhibits the Warburg effect and the invasion of laryngeal cancer cells
10.3760/cma.j.issn.1673-0860.2019.11.010
- VernacularTitle: 靶向下调Yes相关蛋白抑制喉癌细胞Warburg效应和侵袭能力的研究
- Author:
Xiaomin TANG
1
;
Yuxuan SUN
1
;
Wanju LI
1
;
Hao CHEN
1
;
Guanglun WAN
1
;
Jiaqiang SUN
1
;
Chunchen PAN
1
;
Jingwu SUN
1
Author Information
1. Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital of University of Science and Technology of China, Hefei 230001, China
- Publication Type:Journal Article
- Keywords:
Laryngeal neoplasms;
Yes-associated protein;
Glycolysis;
Epithelial mesenchymal transition;
Neoplasm invasiveness
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2019;54(11):857-862
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the migration and invasion behaviors of Hep-2 after the targeted knockdown of yes-associated protein (YAP).
Methods:Hep-2 cells were knock-downed for YAP by shRNA as YAP-shRNA group, Hep-2 treated with non-specific shRNA as YAP-NC group, and Hep-2 with no treatment as control. Glucose uptake and lactate production in the cells were examined to assess Warburg effect. The migration and invasion behaviors of cells in three groups were observed. The expressions of vimentin and E-cadherin were detected by RT-PCR and Western Blot. The statistical software GraphPad Prism 7.0 was used to analyze significance of data. Two tailed Student′ s t-tests was used to determine significance when only two groups were compared. P values of less than 0.05 was considered statistically significant.
Results:Downregulation of YAP led to a obvious decrease in glucose uptake [(18.51±1.72)%] and lactate production [103.40±8.32] in Hep-2 cells compared with control [(41.20±1.11)% and 743.69±19.49, t=19.20 and 52.33, respectively, both P<0.01] and YAP-NC group [(39.60±0.78)% and 705.22±17.20, t=19.34 and 54.56, respectively, both P<0.01]. Compared with the control group (78.32±4.04) and YAP-NC group (77.28±3.11), the scratch healing ability of Hep-2 cells was significantly decreased in YAP-shRNA group (44.71±4.68). The P value was less than 0.01 (t=9.42 and 10.04). The number of cells with YAP-shRNA (33.30±4.19) passing through compartments was remarkable fewer than the control group (133.71±6.72) and YAP-NC group (126.32±4.21). The P value was less than 0.01 (t=21.96 and 27.13). The expression of E-cadherin protein in cells of YAP-shRNA group (6.16±0.11) was up-regulated compared with control (0.97±0.10, t=35.70, P<0.01) and YAP-NC group (1.13±0.09, t=36.28, P<0.01), while the expression of vimentin protein in cells of YAP-shRNA group (1.08±0.09) was down-regulated compared with control (5.67±0.12, t=29.91, P<0.01) and YAP-NC group (5.51±0.12, t=29.04, P<0.01).
Conclusions:The down-regulation of YAP in Hep-2 inhibits the migration and invasion of cells via suppressing Warburg and EMT program.
