Rab7a通过Hippo信号通路促进食管癌细胞演进OA
Rab7a promoted the progression of esophageal cancer cells through the Hippo sig-naling pathway
目的 探讨Ras相关结合蛋白7A(Ras-associated binding protein 7A,Rab7a)对食管癌细胞增殖、迁移能力的影响及与上皮-间质转化(epithelial-mesenchymal transition,EMT)之间的关系,并解析其可能的机制.方法 通过生物信息学分析Rab7a在食管癌中的表达和预后,并利用免疫组化染色对其在食管癌及正常食管黏膜组织中的表达差异加以验证.采用小分子干扰RNA(small interfering RNA,siRNA)技术沉默食管癌TE-1细胞Rab7a后,通过实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)和蛋白免疫印迹(Western blot,WB)法验证敲减效率.将食管癌TE-1细胞分为以下几组:空白对照组(control 组)、空载体对照组(si-NC)、Rab7a 沉默组(si-Rab7a).CCK-8、EdU、克隆形成实验评估Rab7a表达对TE-1细胞增殖能力的影响,细胞划痕、Transwell、Hoechst33342实验分析敲减Rab7a后对TE-1细胞迁移、侵袭及凋亡的影响.通过基因富集分析筛选Rab7a调控食管癌及EMT的关键信号通路,并利用WB实验进行验证.结果 生物信息学分析显示,Rab7a在食管癌中高表达且与患者的预后密切相关.免疫组化染色结果进一步证实,在食管癌组织中,Rab7a表达上调.细胞学实验结果显示,沉默Rab7a表达抑制TE-1细胞增殖、迁移和侵袭,并促进细胞凋亡.富集分析显示Hippo信号通路是Rab7a在食管癌中调控的关键信号通路之一,WB实验进一步证实敲减Rab7a能够抑制Hippo信号通路p-YAP1、YAP1、CTGF和survivin的表达(P<0.01).此外,沉默Rab7a后食管癌TE-1细胞EMT相关蛋白E-cadherin表达上调,而vimentin和Snail表达水平显著降低(P<0.01);Rab7a敲减组凋亡相关蛋白BCL2表达降低而Bax表达上调(P<0.01).结论 Rab7a通过Hippo信号通路调控EMT进程,进而促进食管癌细胞的增殖、迁移及侵袭.
Objective To investigate the effects of Ras-associated binding protein 7A(Rab7a)on the prolifera-tion and migration of esophageal cancer cells,its relationship with epithelial-mesenchymal transition(EMT),and the underlying mechanisms.Methods Bioinformatics analyses were performed to evaluate Rab7a expression and its prog-nostic value in esophageal cancer,followed by immunohistochemical validation in cancer tissues and normal esopha-geal mucosa.Rab7a was silenced in TE-1 cells using small interfering RNA(siRNA),and knockdown efficiency was verified by real-time quantitative PCR(RT-qPCR)and Western blotting(WB).Cells were divided into a control group,a negative control group(si-NC),and a Rab7a-silenced group(si-Rab7a).CCK-8,EdU,and colony forma-tion assays were conducted to assess cell proliferation.Wound-healing,Transwell,and Hoechst 33342 assays were performed to evaluate migration,invasion,and apoptosis.Gene set enrichment analysis was used to identify key sig-naling pathways regulated by Rab7a in esophageal cancer and EMT,and the results were validated by WB.Results Bioinformatics analyses showed that Rab7a was highly expressed in esophageal cancer and was closely associated with patient prognosis.Immunohistochemistry further confirmed upregulated Rab7a expression in esophageal cancer tis-sues.Functional assays demonstrated that Rab7a silencing inhibited proliferation,migration,and invasion of TE-1 cells and promoted apoptosis.Enrichment analysis identified the Hippo signaling pathway as one of the key pathways regulated by Rab7a.WB further demonstrated that Rab7a knockdown reduced the expression of p-YAP1,YAP1,CTGF,and survivin(P<0.01).In addition,Rab7a silencing upregulated E-cadherin expression while significantly downregulating vimentin and Snail(P<0.01).The expression of the anti-apoptotic protein BCL2 decreased,whereas that of the pro-apoptotic protein Bax increased(P<0.01).Conclusion Rab7a regulated EMT through the Hippo sig-naling pathway,thereby promoting the proliferation,migration,and invasion of esophageal cancer cells.
田燕玲;刘博;吕洋
河北北方学院研究生学院,张家口 075000河北北方学院附属第一医院病理科,张家口 075000河北北方学院病理教研室,张家口 075000
医药卫生
食管癌Rab7a生物信息学Hippo信号通路
esophageal cancerRab7abioinformaticsHippo signaling pathway
《临床与实验病理学杂志》 2026 (3)
325-333,341,10
河北省重点研发计划项目卫生健康创新专项(22377765D)、河北省2024年度医学科学研究指导性课题(20240286) Key Research and Development Program of Hebei Province,Health Innovation Special Project(22377765D)2024 Guiding Project for Medical Science Research in Hebei Province(20240286)
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