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

发布时间：2018-01-27 15:33

本文关键词： 子宫内膜癌人附睾蛋白侵袭转染实时定量聚合酶链反应人　出处：《解剖学报》2017年06期 　论文类型：期刊论文

【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(pc DNA 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pc DNA 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pc DNA 3.1/Myc-His-HE4后HEC-1B细胞中HE4 mRNA表达水平显著升高(P0.05),细胞增殖活性、穿膜细胞数明显升高(P0.05);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(P0.05),细胞增殖活性、穿膜细胞数明显降低(P0.05);而阴性对照组与正常对照组之间比较差异无统计学意义(P0.05);HE4过表达组的移植性肿瘤体积和重量明显大于其对照组(P0.05),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(P0.05)。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。
[Abstract]:Objective to investigate the effect of overexpression of human epididymal protein 4 (HE4) on the proliferation of endometrial carcinoma (EC) cells. Methods the overexpression plasmid of HE4 was constructed to construct the pc DNA 3.1% Myc-His-HE4 and HE4 specific small interference RNAs. SiRNA-HE4). Two EC cell lines were transfected into HEC-1B(HE4 overexpression group and Ark2(HE4 low expression group, respectively. HEC-1B cells transfected with empty vector PC DNA 3.1% Myc-His were used as negative control in HE4 overexpression group. Ark2 cells transfected with nonspecific sequence siRNA were used as negative control in HE4 low expression group. Without transfection treatment, the normal cultured EC cell line was the normal control group. The expression level of HE4 mRNA was detected by real-time quantitative polymerase chain reaction real-time PCR. In vitro migration and invasion experiments were used to detect the migration and invasion ability of cells. The volume of tumor formation in each group was compared with that of normal control group and negative control group. The results were compared with those of normal control group and negative control group. After transfection of PC DNA 3.1% Myc-His-HE4, the expression of HE4 mRNA in HEC-1B cells increased significantly. The number of perforating cells increased significantly (P 0.05). Compared with the normal control group and the negative control group, the expression of HE4 mRNA in Ark2 cells after siRNA-HE4 transfection was significantly lower than that in the normal control group and negative control group (P 0.05). The cell proliferation activity and the number of transmembrane cells decreased significantly (P 0.05). There was no significant difference between the negative control group and the normal control group (P 0.05). The volume and weight of transplanted tumor in HE4 overexpression group was significantly larger than that in control group (P0.05). However, the volume and weight of transplanted tumor in the low expression group of HE4 were significantly smaller than that in the control group (P 0.05). Conclusion overexpression of HE4 can increase the proliferation, migration and invasion of EC cells. Promoting tumor formation; Down-regulation of HE4 gene expression can significantly inhibit the proliferation, migration and invasion of EC cells, and inhibit tumor growth.
【作者单位】：承德医学院附属医院妇科;承德医学院附属医院产科;承德医学院附属医院检验科;
【基金】：河北省人口和计划生育委员会科技研究计划项目(2012-A25) 承德市科技支撑计划项目(2013-2044)
【分类号】：R737.33
【正文快照】： 子宫内膜癌(endometrial cancer,EC)是发生于子宫内膜的一组上皮恶性肿瘤,为女性3大生殖系统恶性肿瘤之一,具有较高的发病率和死亡率。前期临床研究表明,近年发现的新型肿瘤标志物人附睾蛋白4(human epididymal protein 4,HE4)对EC的诊断、疾病进展及预后评估中具有重要作用[1

【相似文献】