IKBKE沉默对食管鳞癌细胞生物学行为的影响  

Effects of IKBKE silencing on the biological behavior of esophageal squamous cell carcinoma cells

作　　者：王佳佳[1] 王军[2] 姜黄 WANG Jiajia;WANG Jun;JIANG Huang(Department of Pathology,The Fifth Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou People′s Hospital,Zhengzhou 450003,China;Department of Gynecology,The Fifth Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou People′s Hospital,Zhengzhou 450003,China)

机构地区：[1]河南中医药大学第五临床医学院郑州人民医院病理科,郑州450003 [2]河南中医药大学第五临床医学院郑州人民医院妇科,郑州450003

出　　处：《天津医科大学学报》2024年第4期316-322,共7页Journal of Tianjin Medical University

基　　金：河南省医学科技攻关计划项目(LHGJ20210706)。

摘　　要：目的:探讨核因子(NF)κB激酶亚单位ε抑制剂(IKBKE)在食管鳞状细胞癌(ESCC)中的表达及其沉默对KYSE-30细胞生物学行为的影响。方法:收集自2020年6月至2023年6月开展ESCC根治术的30例患者的癌组织以及癌旁组织,并分为ESCC组(n=30)与对照组(n=30)。运用实时荧光定量聚合酶链反应(qRT-PCR)与蛋白质印迹法(WB)检测ESCC组及对照组中的IKBKE mRNA与蛋白水平,并分析ESCC组中的IKBKE mRNA以及蛋白水平与ESCC患者临床病理特征(性别、年龄与肿瘤的TNM分期、分化、浸润深度及是否转移)的关系。将培养的ESCC细胞系KYSE-30细胞随机分为空白对照组(无任何处理)、NC-IKBKE组(转染NC-siRNA)、si-IKBKE组(转染IKBKE-siRNA)。CCK-8法检测KYSE-30细胞增殖能力;流式细胞术检测KYSE-30细胞凋亡率;划痕法检测KYSE-30细胞迁移能力;Transwell法检测KYSE-30细胞侵袭能力;采用qRT-PCR及WB检测KYSE-30细胞中IKBKE mRNA及蛋白水平;采用WB检测KYSE-30细胞中NF-κB通路关键蛋白NF-κB p65、磷酸化核因子κB抑制蛋白α(p-IκBα)表达水平。结果:ESCC组IKBKE的mRNA及蛋白表达水平均显著高于对照组(t=9.769、12.960,均P=0.000);IKBKE在ESCC组的mRNA及蛋白水平与ESCC的肿瘤的分化程度、TNM分期及转移有关(t=2.070~2.829,均P<0.05),而其与性别、年龄、浸润深度无关(均P>0.05);与NC-IKBKE组相比,si-IKBKE组KYSE-30细胞中的IKBKE的mRNA以及蛋白水平均显著降低(F=77.244、78.436,均P=0.000);成功沉默IKBKE表达的KYSE-30细胞系后,与NC-IKBKE组相比,si-IKBKE组细胞的48、72 h细胞增殖能力及24、48 h细胞迁移率均显著下降(F=12.355~44.755,均P<0.05),与NC-IKBKE组比较,si-IKBKE组48 h细胞侵袭抑制率、细胞凋亡率均明显增加(F=155.436、46.360,均P=0.000);si-IKBKE组的NF-κB p65、p-IκBα的蛋白表达水平均显著低于NC-IKBKE组(F=139.206、55.551,均P=0.000)。结论:ESCC组织中的IKBKE表达水平显著高于癌旁组织,且与肿瘤的分化程度、TNM�Objective:To investigate the expression of nuclear factor(NF)-κB kinase subunitεinhibitor(IKBKE)in esophageal squamous cell carcinoma(ESCC)and the effect of IKBKE silencing on the biological behavior of KYSE-30 cell.Methods:The cancer tissues and adjacent tissues of 30 patients who underwent radical ESCC operation in our hospital from June 2020 to June 2023 were collected and divided into ESCC group(n=30)and control group(n=30).Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)and Western blotting(WB)were used to detect IKBKE mRNA and protein levels in ESCC group and control group.The relationship between IKBKE mRNA and protein levels and clinicopathological characteristics of ESCC patients(gender,age,TNM stage,differentiation,depth of invasion and metastasis)was analyzed.The cultured ESCC cell line KYSE-30 cells were randomly divided into blank control group(without any treatment),NC-IKBKE group(transfected with NC-siRNA)and si-IKBKE group(transfected with IKBKE-siRNA).CCK-8 assay was used to detect the proliferation of KYSE-30 cells.The apoptosis rate of KYSE-30 cells was detected by flow cytometry.The migration ability of KYSE-30 cells was detected by scratch assay.Transwell assay was used to detect the invasion ability of KYSE-30 cells.QRT-PCR and WB were used to detect the mRNA and protein levels of IKBKE in KYSE-30 cells.WB was used to detect the expression levels of NF-κB pathway key protein including NF-κB p65 and phosphorylated nuclear factor-κb inhibitor proteinα(p-IκBα)in KYSE-30 cells.Results:The expression levels of IKBKE mRNA and protein in ESCC group were significantly higher than those in control group(t=9.769,12.960,all P=0.000).The mRNA and protein levels of IKBKE in ESCC group were related to the degree of tumor differentiation,TNM stage and metastasis(t=2.070-2.829,all P<0.05),but not related to gender,age and depth of invasion(all P>0.05).Compared with the NC-IKBKE group,the mRNA and protein levels of IKBKE in KYSE-30 cells of the si-IKBKE group were significantly d

关 键 词：核因子κB激酶亚单位ε抑制剂 食管鳞癌 KYSE-30细胞 增殖 迁移 侵袭 

分 类 号：R735.1[医药卫生—肿瘤]