解剖学报

解剖学报 ›› 2017, Vol. 48 ›› Issue (6): 704-709.doi: 10.16098/j.issn.0529-1356.2017.06.012

• 生物工程学 • 上一篇    下一篇

人附睾蛋白4过表达对子宫内膜癌细胞增殖、侵袭能力及肿瘤形成的影响

崔彭华1 张玉娟1* 邵雪斋1 王会民2 邢恩鸿3   

  1. 1.承德医学院附属医院妇科; 2.产科; 3.检验科，河北 承德 067000
  • 收稿日期:2017-07-24 修回日期:2017-08-17 出版日期:2017-12-06 发布日期:2017-12-06
  • 通讯作者: 张玉娟 E-mail:cuipenghua47@163.com
  • 基金资助:
    河北省人口和计划生育委员会科技研究计划项目;承德市科技支撑计划项目（2013-2044）。

Effect of overexpression of human epididymal protein 4 on proliferation,invasion and tumorigenesis of endometrial carcinoma cells

CUI Peng-hua1 ZHANG Yu-juan 1* SHAO Xue-zhai1 WANG Hui-min2 XING En-hong3   

  1. 1.Department of Gynaecology; 2.Department of Obstetrics; 3.Department of Clinical Laboratory, Affiliated Hospital of Chengde Medical College, Hebei Chengde 067000, China
  • Received:2017-07-24 Revised:2017-08-17 Online:2017-12-06 Published:2017-12-06
  • Contact: ZHANG Yu-juan E-mail:cuipenghua47@163.com

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摘要：

目的 探讨人附睾蛋白4（HE4）过表达对子宫内膜癌（EC）细胞增殖、侵袭能力及肿瘤形成的影响。 方法 构建HE4过表达质粒载体（pcDNA 3.1/Myc-His-HE4）和HE4特异性小干扰RNA（siRNA）表达质粒载体（siRNA-HE4），分别转染HEC-1B（HE4过表达组）和Ark2（HE4低表达组）两个EC细胞系；以空载体pcDNA 3.1/Myc-His转染的HEC-1B细胞为HE4过表达组的阴性对照，以非特异性序列siRNA转染的Ark2细胞为HE4低表达组的阴性对照；不进行转染处理、正常培养的EC细胞系为正常对照组。转染后，采用实时定量聚合酶链反应（Real-time PCR）法检测HE4 mRNA表达水平，MTT比色法测定细胞增殖活性，体外迁移和侵袭实验检测细胞的迁移和侵袭能力。构建移植性肿瘤小鼠模型，比较各组动物的形成瘤体积。 结果 与正常对照组和阴性对照组的HEC-1B细胞比，转染pcDNA 3.1/Myc-His-HE4后HEC-1B细胞中HE4 mRNA表达水平显著升高（P＜0.05），细胞增殖活性、穿膜细胞数明显升高（P＜0.05）；与正常对照组和阴性对照组的Ark2细胞比，转染siRNA-HE4后Ark2细胞中HE4 mRNA表达水平显著降低（P＜0.05），细胞增殖活性、穿膜细胞数明显降低（P＜0.05）；而阴性对照组与正常对照组之间比较差异无统计学意义（P＞0.05）；HE4过表达组的移植性肿瘤体积和重量明显大于其对照组（P＜0.05），而HE4低表达组的移植性肿瘤体积和重量明显小于其对照组（P＜0.05）。 结论 HE4过表达可导致EC细胞的增殖、迁移及侵袭能力增强，促进肿瘤形成；下调HE4基因表达可明显抑制EC细胞的增殖、迁移和侵袭，并抑制肿瘤生长。

关键词: 子宫内膜癌, 人附睾蛋白4, 侵袭, 转染, 实时定量聚合酶链反应, 人

Abstract:

Objective To investigate the effect of overexpression of human epididymal protein 4 (HE4) on proliferation, invasion and tumorigenesis of endometrial carcinoma (EC) cells. Methods HE4 overexpression vector (pcDNA 3.1/Myc-His-HE4) and HE4 specific small interfering RNA (siRNA-HE4) were constructed and respectively transfected into HEC-1B (HE4 overexpression group) and Ark2 (HE4 low expression group), the two EC cell lines. HEC-1B cells transfected with empty vector pcDNA 3.1/Myc-His were used as negative control for the HE4 overexpression group. Ark2 cells transfected with nonspecific sequence siRNA were used as negative control for the HE4 low expression group. The EC cell line without transfection and normal culture served as a normal control group. After transfection, the expression levels of HE4 mRNA were detected by quantitative real-time PCR (Real-time PCR), and cell proliferation was measured by methylthiazoletrazolium (MTT) assay. Migration and invasion of cells were examined by migration and invasion experiments in vitro. The transplanted tumor mice model were constructed and the tumor volume were compared. Results Compared with the HEC-1B cells in the normal control group and negative control group, the expression level of HE4 mRNA in the HEC-1B cells increased significantly after transfection with pcDNA 3.1/Myc-His-HE4 (P<0.05), and the cell proliferation activity, transmembrane cell number increased obviously (P<0.05). Compared with the Ark2 cells in the negative control group and the normal control group, the expression of HE4 mRNA in the Ark2 cells decreased significantly after transfection with siRNA-HE4 (P<0.05), the cell proliferation activity and transmembrane cell number also significantly decreased (P<0.05). While, no significant difference between the negative control group and the normal control group was found (P> 0.05). The volume and weight of transplanted tumors in the HE4 overexpression group were significantly higher than those in the other two control groups (P<0.05). And the volume and weight of transplanted tumors in the HE4 low expression group were significantly smaller than those in the other two control groups (P<0.05). Conclusion Overexpression of HE4 could lead to the proliferation, migration and invasion of EC cells, and promote tumor formation. Downregulation of HE4 gene expression could inhibit the proliferation, migration and invasion of EC cells and inhibit the tumor growth.

Key words: Endometrial cancer, Human epididymal protein 4, Invasion, Transfection, Real-time PCR, Human