IPO5基因表达对食管癌恶性进展的促进作用

李美玉 李晓飞 张天爱 郭莲怡

李美玉, 李晓飞, 张天爱, 郭莲怡. IPO5基因表达对食管癌恶性进展的促进作用[J]. 中国肿瘤临床. doi: 10.12354/j.issn.1000-8179.2022.20220590
引用本文: 李美玉, 李晓飞, 张天爱, 郭莲怡. IPO5基因表达对食管癌恶性进展的促进作用[J]. 中国肿瘤临床. doi: 10.12354/j.issn.1000-8179.2022.20220590
Meiyu Li, Xiaofei Li, Tianai Zhang, Lianyi Guo. Importin5 gene expression promotes malignant progression of esophageal cancer[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY. doi: 10.12354/j.issn.1000-8179.2022.20220590
Citation: Meiyu Li, Xiaofei Li, Tianai Zhang, Lianyi Guo. Importin5 gene expression promotes malignant progression of esophageal cancer[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY. doi: 10.12354/j.issn.1000-8179.2022.20220590

IPO5基因表达对食管癌恶性进展的促进作用

doi: 10.12354/j.issn.1000-8179.2022.20220590
基金项目: 本文课题受辽宁省教育厅基金青年项目(编号:JYTQN2020031)资助
详细信息
    作者简介:

    李美玉:专业方向为消化内科

    通讯作者:

    郭莲怡 angel_gly@163.com

Importin5 gene expression promotes malignant progression of esophageal cancer

Funds: This work was supported by the Foundation of Liaoning Education Department Youth Project (No.JYTQN2020031)
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  • 摘要:   目的  探讨importin5(IPO5)基因的表达对食管癌(esophageal cancer, EC)进展的影响以及相关作用机制。  方法  收集2020年9 月至2022 年4 月50 例于锦州医科大学附属第一医院确诊为EC的患者的资料及手术标本,免疫组化验证IPO5在癌组织与癌旁组织中的表达情况,评估IPO5与患者临床信息的相关性;通过慢病毒转染构建IPO5基因沉默EC细胞模型;通过 CCK-8实验、Transwell侵袭实验、细胞划痕实验检测细胞增殖、侵袭及迁移能力;应用流式细胞术检测细胞周期,Western blot实验检测细胞周期相关蛋白的表达及RAS通路相关蛋白的表达。构建裸鼠皮下成瘤模型,免疫组化验证Ki-67表达。  结果  临床实验中,IPO5在癌组织的表达明显高于癌旁组织(P<0.01),并与肿瘤大小、分期及分化程度呈正相关(P<0.05)。IPO5在EC细胞中的表达高于正常食管细胞,尤其是ECA109及OE33细胞(P<0.01)。与阴性对照组相比,IPO5干扰组细胞增殖能力减弱(P<0.05)、侵袭能力减弱(P<0.01)。IPO5基因沉默后EC细胞生长周期阻滞于G2期,细胞周期相关蛋白表达减少。sh-IPO5干扰后RAS-ERK通路下游蛋白表达水平下降(P<0.05)。裸鼠成瘤实验证实敲除IPO5后肿瘤缩小(P<0.05),Ki-67表达减少(P<0.01)。  结论  IPO5在EC组织中高表达,抑制IPO5的表达对于EC细胞的生长和迁移存在显著影响,IPO5通过激活RAS-ERK信号通路促进EC的进展。

     

  • 图  1  EC中IPO5的表达情况

    A~C:EC组织与正常食管组织之间差异表达基因的火山图,IPO5在EC组织中呈现高表达。D:EC组织中IPO5高表达;E:EC癌旁组织中IPO5低表达,F:依据免疫组化评分计算出IPO5在癌组织中表达升高。G:qRT-PCR分析IPO5在ECA109和OE33细胞中表达高于HEEC细胞;H:Western blot实验显示sh-IPO5可以明显下调EC细胞中IPO5的表达。注:*:P<0.05;**:P<0.01

    图  2  IPO5影响EC细胞的侵袭、迁移、增殖能力

    A:细胞划痕实验显示在24h时sh-IPO5组闭合率低于sh-NC组;B:Transwell侵袭实验证明干扰IPO5表达可使细胞侵袭能力减弱;C:CCK-8分析sh-IPO5可以明显减低ECA109和OE33细胞活力。注:*:P<0.05;**:P<0.01

    图  3  IPO5影响食管癌的细胞周期及肿瘤大小

    A、B:流式细胞术检测ECA109细胞转染sh-IPO5后G2期细胞比例增多;C、D:流式细胞术检测OE33细胞转染sh-IPO5后G2期细胞比例增多。E:Western blot分析显示敲低IPO5基因可使CDK4、CDK6、CyclinD1表达减少。 F:裸鼠成瘤实验显示sh-IPO5组裸鼠生长速度明显小于对照组;G:免疫组化实验显示阳性组与阴性对照组相比Ki67表达明显降低。*:P<0.05;**:P<0.01

    表  1  临床数据与IPO5表达之间的关系 例

    项目例数IPO5低表达IPO5高表达χ2P
    性别0.1090.741
     男271413
     女231310
    年龄(岁)0.2160.642
     ≤6022148
     >60281612
    肿瘤大小(cm)5.5470.019
     ≤3.9281711
     >3.922616
    TNM分期4.2760.039
     Ⅰ~Ⅱ362214
     Ⅲ~Ⅳ14410
    淋巴结转移2.8030.094
     有1239
     无382018
    分化程度6.3880.041
     低817
     中291415
     高1394
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  • 收稿日期:  2022-04-22
  • 录用日期:  2022-07-19
  • 修回日期:  2022-07-06
  • 网络出版日期:  2022-08-17

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