RESUMEN
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.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/metabolismo , Factores de Transcripción/genética , Proteínas Adaptadoras Transductoras de Señales/biosíntesis , Cadherinas/biosíntesis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , Humanos , Neoplasias Laríngeas/patología , Invasividad Neoplásica , ARN Interferente Pequeño/genética , Factores de Transcripción/biosíntesis , Vimentina/biosíntesis , Proteínas Señalizadoras YAPRESUMEN
From 1978 through 1981, three teams of ophthalmologists from the United States have traveled to the People's Republic of China under the auspices of the World Eye Foundation. The purpose of these visits has been to open an avenue of exchange between American and Chinese ophthalmologists. The US ophthalmologists delivered lectures and participated in discussion groups with the Chinese ophthalmologists. They had the opportunity to participate in surgical teaching cases, including some using acupuncture anesthesia. They also had the opportunity to develop ongoing working and research relationships with many Chinese ophthalmologists. As a result of these working relationships, some of these Chinese ophthalmologists have had the opportunity to visit and study in the United States.
