摘要
目的探讨泛素结合酶E2B(UBE2B)在食管癌中的表达及其与上皮-间质转化和转移的关系。方法利用生物信息分析癌症基因组图谱数据库中食管癌UBE2B mRNA表达水平及其对患者生存的影响。采用免疫组化法检测食管癌组织和正常组织中UBE2B蛋白的表达水平。将Eca-109细胞分别转染特异性靶向UBE2B的siRNA以敲减UBE2B的表达(si-UBE2B组)和非靶向对照siRNA(si-control组)。采用Transwell小室实验检测细胞迁移、侵袭能力。采用Western blot法检测细胞UBE2B及上皮-间质转化标志蛋白[上皮性钙黏附蛋白(E-cadherin)、神经性钙黏附蛋白(N-cadherin)、Slug和Snail]表达水平。结果生物信息分析结果显示,与正常组织比较,食管癌组织中UBE2B mRNA表达水平较高(P<0.01)。生存分析结果显示UBE2B高表达组患者总体生存率明显低于UBE2B低表达组(P<0.01)。组织芯片免疫组化结果显示,食管癌组织中UBE2B蛋白阳性表达率为76.47%,高于正常组织的29.41%,差异有统计学意义(P<0.01)。Transwell小室实验提示与si-control组比较,si-UBE2B组迁移细胞数和侵袭细胞数均明显降低(均P<0.01)。与si-control组比较,si-UBE2B组细胞上皮标志物E-cadherin蛋白明显升高,而间质标志物N-cadherin、Slug和Snail均明显降低,差异均有统计学意义(均P<0.01)。结论UBE2B在食管癌中异常高表达,并且与不良预后有关。敲低UBE2B通过抑制食管癌细胞上皮-间质转化抑制细胞迁移、侵袭能力。
Objective To explore the expression of UBE2B in esophageal cancer and its relationship with epithelialmesenchymal transition(EMT)and metastasis.Methods The data of UBE2B mRNA expression levels in esophageal cancer and its impact on survival prognosis were obtained from the Cancer Genome Atlas(TCGA)database.Immunohistochemistry was used to detect the protein expression level of UBE2B in esophageal cancer and adjacent normal tissues.Eca-109 cells were transfected with specific UBE2B targeting siRNA(si-UBE2B group)or untargeted control siRNA(si-control group).Transwell assay was performed to detect cell migration and invasion ability;Western blot was conducted to assess the effect of low expression of UBE2B on EMT-related marker proteins.Results Bioinformatics analysis showed that the expression level of UBE2B mRNA in esophageal cancer tissues was higher than that in normal tissues(P<0.01).Survival analysis results showed that the overall survival rate of patients with high UBE2B expression group was significantly lower than that of patients with low UBE2B expression group(P<0.01).Immunohistochemical results of tissue microarray showed that the positive expression rate of UBE2B protein in esophageal cancer tissues was 76.47%,higher than in normal tissues(29.41%,P<0.01).Transwell chamber assay showed that the number of migrating cells and invading cells in si-UBE2B group were significantly lower than those in si-control group(both P<0.01).Compared with si-control group,the epithelial marker Ecadherin protein was significantly increased in si-UBE2B group,while the interstitial markers N-cadherin,Slug and Snail were significantly decreased(all P<0.01).Conclusion UBE2B is abnormally highly expressed in esophageal cancer and is significantly related to poor prognosis.Knockdown of UBE2B inhibits cell migration and invasion by inhibiting the EMT of esophageal cancer cells.
作者
施谷平
孙宇
SHI Guping;SUN Yu(The Second Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310053,China;不详)
出处
《浙江医学》
CAS
2022年第14期1486-1490,I0006,共6页
Zhejiang Medical Journal
基金
嘉兴市科技计划项目(2019AD32204)。
关键词
食管癌
泛素结合酶E2B
上皮-间质转化
迁移
侵袭
Esophageal cancer
Ubiquitin conjugating enzyme E2B
Epithelial-mesenchymal transition
Migration
Invasion