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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年04期

页码:

316-322

栏目:

肿瘤疾病专题

出版日期:

2024-07-10

文章信息/Info

Title:

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

文章编号:

1006-8147(2024)044-0316-07

作者:

王佳佳¹; 王军²; 姜黄¹

（河南中医药大学第五临床医学院郑州人民医院 1.病理科；2.妇科，郑州 450003）

Author(s):

WANG Jiajia¹; WANG Jun²; JIANG Huang¹

（1.Department of Pathology；2.Department of Gynecology，The Fifth Clinical Medical College of Henan University of Chinese Medicine， Zhengzhou People′s Hospital，Zhengzhou 450003，China）

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

Keywords:

nuclear factor-κB kinase subunit ε inhibitor; esophageal squamous cell carcinoma; KYSE-30 cell; proliferation; migration; invasion

分类号:

R735.1

DOI:

10.20135/j.issn.1006-8147.2024.04.0316

文献标志码:

A

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分期及转移有关，沉默IKBKE可显著抑制ESCC细胞的增殖、迁移以及侵袭能力，促进细胞凋亡，其机制可能与IKBKE调控NF－κB通路有关。

Abstract:

Objective： To investigate the expression of nuclear factor（NF）-κB kinase subunit ε inhibitor（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 protein α（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 decreased（F=77.244，78.436，all P=0.000）. After successfully silencing IKBKE expression in KYSE-30 cell line，compared with NC-IKBKE group the 48 h and 72 h cell proliferation ability and 24 h and 48 h cell migration rate in the si-IKBKE group were significantly decreased（F=12.355-44.755，all P<0.05）. Compared with the NC-IKBKE group the cell invasion inhibition rate and apoptosis rate in the si-IKBKE group were significantly increased at 48 h（F=155.436， 46.360，all P=0.000）. The protein expression levels of NF-κB p65 and p-IκBα in the si-IKBKE group were significantly lower than those in the NC-IKBKE group（F=139.206，55.551，all P=0.000）. Conclusion：The expression level of IKBKE in ESCC tissues is significantly higher than that in adjacent tissues，which is related to the degree of tumor differentiation，TNM stage and metastasis. IKBKE silencing can significantly inhibit the proliferation，migration and invasion of ESCC cells and promote cell apoptosis，which may be related to IKBKE regulating the NF-κB pathway.

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相似文献/References:

备注/Memo

备注/Memo:

基金项目：河南省医学科技攻关计划项目（LHGJ20210706）
作者简介：王佳佳（1987-），女，学士，主治医师，研究方向：消化道肿瘤；
通信作者：姜黄，E-mail：jiaahuang@126.com。

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更新日期/Last Update: 2024-07-10