BACKGROUND Uncoupling protein 1(UCP1)plays a pivotal role in modulating energy expen-diture and maintaining metabolic homeostasis within brown and beige adipo-cytes.It has also been implicated in tumorigenesis.AIM To ...BACKGROUND Uncoupling protein 1(UCP1)plays a pivotal role in modulating energy expen-diture and maintaining metabolic homeostasis within brown and beige adipo-cytes.It has also been implicated in tumorigenesis.AIM To investigate the expression and function of UCP1 in gastric cancer(GC).METHODS UCP1 protein expression in 211 GC tissues was examined using immunohisto-chemistry.Bisulfite sequencing PCR(BSP)was used to detect the methylation status of the UCP1 promoter in GC cell lines and tissues.The relationship between UCP1 expression and clinicopathological parameters was analyzed.CCK8,scratch,transwell,and flow cytometry assays were carried out to analyze the proliferation,migration,invasion,and apoptosis of GC cell lines after knockdown or overexpression of UCP1 in vitro.A nude mouse tumor xenograft model was used to investigate the function of UCP1 in vivo.RNA sequencing,Kyoto Ency-clopedia of Genes and Genomes analysis,and Rap1 pull-down assays were performed to identify the pathway associated with UCP1.RESULTS Loss of UCP1 was significantly associated with gender,poor differentiation,and advanced TNM stage of GC.Hypermethylation of UCP1 was confirmed in GC cells and tumor tissues by BSP.Overexpression of UCP1 suppressed GC cell proliferation,migration,and invasion,and it promoted apoptosis in vitro.UCP1 overexpression also suppressed GC tumor growth in vivo.Moreover,overexpression of UCP1 in GC cells resulted in a significant decrease in active Rap1 protein levels,whereas downregulation of UCP1 markedly enhanced Rap1 activity.CONCLUSION UCP1 downregulation in GC through promoter hypermethylation is related to the progression of GC,indicating that UCP1 plays a role as a tumor suppressor in GC.It regulates Rap1 signaling and may be a potential therapeutic target in GC.展开更多
目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;...目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。展开更多
基金supported by the National Natural Science Foundation of China(No.82160051,32100908)the Jiangxi Provincial Natural Science Foundation(No.20232BAB206018)+2 种基金the Science and Technology Research Project in the Education Department of Jiangxi Province,China(No.GJJ2200904)the Ph.D.Start-up Research Fund in Jiangxi University of Chinese Medicine(No.2020BSZR009)the Discipline of Chinese and Western Integrative Medicine in Jiangxi University of Chinese Medicine(No.zxyylxk20220103)。
基金Supported by the Nanjing Health Science and Technology Development Fund,No.YKK24223.
文摘BACKGROUND Uncoupling protein 1(UCP1)plays a pivotal role in modulating energy expen-diture and maintaining metabolic homeostasis within brown and beige adipo-cytes.It has also been implicated in tumorigenesis.AIM To investigate the expression and function of UCP1 in gastric cancer(GC).METHODS UCP1 protein expression in 211 GC tissues was examined using immunohisto-chemistry.Bisulfite sequencing PCR(BSP)was used to detect the methylation status of the UCP1 promoter in GC cell lines and tissues.The relationship between UCP1 expression and clinicopathological parameters was analyzed.CCK8,scratch,transwell,and flow cytometry assays were carried out to analyze the proliferation,migration,invasion,and apoptosis of GC cell lines after knockdown or overexpression of UCP1 in vitro.A nude mouse tumor xenograft model was used to investigate the function of UCP1 in vivo.RNA sequencing,Kyoto Ency-clopedia of Genes and Genomes analysis,and Rap1 pull-down assays were performed to identify the pathway associated with UCP1.RESULTS Loss of UCP1 was significantly associated with gender,poor differentiation,and advanced TNM stage of GC.Hypermethylation of UCP1 was confirmed in GC cells and tumor tissues by BSP.Overexpression of UCP1 suppressed GC cell proliferation,migration,and invasion,and it promoted apoptosis in vitro.UCP1 overexpression also suppressed GC tumor growth in vivo.Moreover,overexpression of UCP1 in GC cells resulted in a significant decrease in active Rap1 protein levels,whereas downregulation of UCP1 markedly enhanced Rap1 activity.CONCLUSION UCP1 downregulation in GC through promoter hypermethylation is related to the progression of GC,indicating that UCP1 plays a role as a tumor suppressor in GC.It regulates Rap1 signaling and may be a potential therapeutic target in GC.
文摘目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。
