目的探讨中心体蛋白78(centrosomal protein 78,CEP78)对内皮细胞生物学行为的影响及其与冠心病(coronary heart disease,CHD)的关系。方法通过Gene Expression Omnibus(GEO)数据库检索CHD相关数据集,利用StataSE 15求CEP78的总标准化...目的探讨中心体蛋白78(centrosomal protein 78,CEP78)对内皮细胞生物学行为的影响及其与冠心病(coronary heart disease,CHD)的关系。方法通过Gene Expression Omnibus(GEO)数据库检索CHD相关数据集,利用StataSE 15求CEP78的总标准化平均差(standardized mean difference,SMD)并绘制总受试者操作特征(summary receiver operating characteristic,SROC)曲线。利用单细胞RNA测序分析CEP78在不同细胞中的表达情况。在人脐静脉内皮细胞株(EA.hy926)中构建过表达CEP78(OE_CEP78)和过表达空载质粒(OE_NC)模型,通过细胞划痕、Transwell、CCK8和细胞凋亡实验验证CEP78和CHD之间的相关性。利用SMD和Spearman相关性分析求CEP78差异共表达基因,通过Metascape数据库对CEP78差异共表达基因进行基因本体(gene ontology,GO)和京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析。结果CEP78在CHD中低表达(总SMD=-0.99,95%CI:-1.79~-0.20,P=0.015),SROC曲线下面积(area under the curve,AUC)为0.77(0.73~0.81),证明CEP78对CHD具有中等诊断能力。单细胞RNA测序分析结果显示,CEP78在正常外周血自然杀伤细胞中高表达。OE_CEP78可以抑制EA.hy926细胞的增殖、迁移和凋亡。KEGG通路富集分析显示CEP78差异正相关基因富集在FOXO信号通路。结论CEP78在CHD中低表达,对CHD具有中等诊断能力,并通过抑制内皮细胞的增殖、迁移和凋亡来抑制CHD的发生发展。展开更多
Introduction:Acral melanoma(AM)is the predominant subtype of cutaneous melanoma in Asian populations,characterized by more aggressive clinical features and limited neoadjuvant therapy response.Centrosomal protein 55 k...Introduction:Acral melanoma(AM)is the predominant subtype of cutaneous melanoma in Asian populations,characterized by more aggressive clinical features and limited neoadjuvant therapy response.Centrosomal protein 55 kDa(CEP55)has been implicated in the pathogenesis of various malignancies,but its role in AM remains undefined.Methods:CEP55 expression in melanoma tissues and cell lines was analyzed by RT-qPCR,Western blotting,and immunohistochemistry(IHC).Databases(GEPIA,Sangerbox,Kaplan-Meier plotter,and TIMER)were used to analyze the expression of CEP55 and its correlation with clinical data of melanoma patients.Functional assays were conducted in vitro and in vivo.RNA sequencing(RNA-seq)and rescue experiments were used to explore underlying mechanisms.Tissue microarrays were used to verify the relationship between CEP55 and immune checkpoints.Results:CEP55 overexpression is associated with Breslow thickness and TNM stage in melanoma tissues and cell lines.CEP55 knockdown inhibited melanoma cell proliferation,migration,and invasion.And CEP55 activated mitogen-activated protein kinase(MAPK)signaling,leading to BRAF inhibitor resistance.Besides,CEP55 overexpression was associated with more favorable responses to immunotherapy in melanoma patients.Conclusions:CEP55 plays a critical role in AM progression and immunotherapy.Targeting CEP55 may be a promising therapeutic strategy for AM.展开更多
The aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with the development of various carcinomas,especially colorectal cancers(CRCs),where adenomatous colorectal polyposis(APC)mutations...The aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with the development of various carcinomas,especially colorectal cancers(CRCs),where adenomatous colorectal polyposis(APC)mutations are the most frequently observed,which limits the anti-tumor efficiency of inhibitors targeting the upstream of Wnt/β-catenin pathway.The anti-tumor activity of the naturally occurring alkaloid cepharanthine(CEP)extracted from the plant Stephania cepharantha Hayata has been reported in various types of tumors.We previously observed that its derivatives inhibited the Wnt/β-catenin signaling in liver cancer;however,the specific mechanism remains unknown.In this study,we confirmed CEP can effectively inhibit APC-mutant CRC cell lines(SW480,SW620,LoVo)through disturbing of the Wnt/β-catenin signaling and elucidated the underlying mechanisms.Here,we demonstrate that CEP attenuates the Wnt/β-catenin signaling by decreasing theβ-catenin,subsequently impeding the proliferation of APC-mutant CRCs.Moreover,CEP inducedβ-catenin transcription inhibition rather than the instability ofβ-catenin protein and mRNA contributes to reduction ofβ-catenin.Taken together,our findings identify CEP as the firstβ-catenin transcriptional inhibitor in the modulation of Wnt/β-catenin signaling and indicate CEP as a potential therapeutic option for the treatment of APC-mutated CRCs.展开更多
文摘目的探讨中心体蛋白78(centrosomal protein 78,CEP78)对内皮细胞生物学行为的影响及其与冠心病(coronary heart disease,CHD)的关系。方法通过Gene Expression Omnibus(GEO)数据库检索CHD相关数据集,利用StataSE 15求CEP78的总标准化平均差(standardized mean difference,SMD)并绘制总受试者操作特征(summary receiver operating characteristic,SROC)曲线。利用单细胞RNA测序分析CEP78在不同细胞中的表达情况。在人脐静脉内皮细胞株(EA.hy926)中构建过表达CEP78(OE_CEP78)和过表达空载质粒(OE_NC)模型,通过细胞划痕、Transwell、CCK8和细胞凋亡实验验证CEP78和CHD之间的相关性。利用SMD和Spearman相关性分析求CEP78差异共表达基因,通过Metascape数据库对CEP78差异共表达基因进行基因本体(gene ontology,GO)和京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析。结果CEP78在CHD中低表达(总SMD=-0.99,95%CI:-1.79~-0.20,P=0.015),SROC曲线下面积(area under the curve,AUC)为0.77(0.73~0.81),证明CEP78对CHD具有中等诊断能力。单细胞RNA测序分析结果显示,CEP78在正常外周血自然杀伤细胞中高表达。OE_CEP78可以抑制EA.hy926细胞的增殖、迁移和凋亡。KEGG通路富集分析显示CEP78差异正相关基因富集在FOXO信号通路。结论CEP78在CHD中低表达,对CHD具有中等诊断能力,并通过抑制内皮细胞的增殖、迁移和凋亡来抑制CHD的发生发展。
基金supported by CAMS Innovation Fund for Medical Sciences(CIFMS)(2024-I2M-C&T-B-089)National Key Research and Development Program(2022YFC2504700,2022YFC2504701,2022YFC2504705)National Natural Science Foundation of China(NSFC81872216).
文摘Introduction:Acral melanoma(AM)is the predominant subtype of cutaneous melanoma in Asian populations,characterized by more aggressive clinical features and limited neoadjuvant therapy response.Centrosomal protein 55 kDa(CEP55)has been implicated in the pathogenesis of various malignancies,but its role in AM remains undefined.Methods:CEP55 expression in melanoma tissues and cell lines was analyzed by RT-qPCR,Western blotting,and immunohistochemistry(IHC).Databases(GEPIA,Sangerbox,Kaplan-Meier plotter,and TIMER)were used to analyze the expression of CEP55 and its correlation with clinical data of melanoma patients.Functional assays were conducted in vitro and in vivo.RNA sequencing(RNA-seq)and rescue experiments were used to explore underlying mechanisms.Tissue microarrays were used to verify the relationship between CEP55 and immune checkpoints.Results:CEP55 overexpression is associated with Breslow thickness and TNM stage in melanoma tissues and cell lines.CEP55 knockdown inhibited melanoma cell proliferation,migration,and invasion.And CEP55 activated mitogen-activated protein kinase(MAPK)signaling,leading to BRAF inhibitor resistance.Besides,CEP55 overexpression was associated with more favorable responses to immunotherapy in melanoma patients.Conclusions:CEP55 plays a critical role in AM progression and immunotherapy.Targeting CEP55 may be a promising therapeutic strategy for AM.
基金funded by the National Natural Science Foundation of China(No.32260159,82360725)the Applied Basic Research Foundation of Yunnan Provincet(202301AS070022)+4 种基金Yunnan University(start-up grant to Y.L.and L.K.)the Yunnan Young&Elite Talents Project(YNWR-QNBJ-2020-084)the Central Guidance on Local Science and Technology Development Fund of Yunnan Province(202207AB110002)National Key R&D Program of China(2019YFE0109200)the central government guides local science and technology development fund(202207AA110007).
文摘The aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with the development of various carcinomas,especially colorectal cancers(CRCs),where adenomatous colorectal polyposis(APC)mutations are the most frequently observed,which limits the anti-tumor efficiency of inhibitors targeting the upstream of Wnt/β-catenin pathway.The anti-tumor activity of the naturally occurring alkaloid cepharanthine(CEP)extracted from the plant Stephania cepharantha Hayata has been reported in various types of tumors.We previously observed that its derivatives inhibited the Wnt/β-catenin signaling in liver cancer;however,the specific mechanism remains unknown.In this study,we confirmed CEP can effectively inhibit APC-mutant CRC cell lines(SW480,SW620,LoVo)through disturbing of the Wnt/β-catenin signaling and elucidated the underlying mechanisms.Here,we demonstrate that CEP attenuates the Wnt/β-catenin signaling by decreasing theβ-catenin,subsequently impeding the proliferation of APC-mutant CRCs.Moreover,CEP inducedβ-catenin transcription inhibition rather than the instability ofβ-catenin protein and mRNA contributes to reduction ofβ-catenin.Taken together,our findings identify CEP as the firstβ-catenin transcriptional inhibitor in the modulation of Wnt/β-catenin signaling and indicate CEP as a potential therapeutic option for the treatment of APC-mutated CRCs.
