TRIM14在子宫内膜癌中的表达及其对子宫内膜癌细胞侵袭和转移的影响
| 作者: |
1贺敏,
2陈勇伟,
1曾康康,
1郑蓉
1 十堰市太和医院(湖北医药学院附属医院)妇产科，湖北 十堰 442700 2 十堰市太和医院(湖北医药学院附属医院)泌尿外科，湖北 十堰 442700 |
| 通讯: |
贺敏
Email: 3036420578@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2019.10.006 |
摘要
目的：研究三结构域蛋白14(tripartite motif-containing 14，TRIM14)在子宫内膜癌组织中的表达与子宫内膜癌患者临床病理特征及预后的关系，并探讨TRIM14对子宫内膜癌细胞侵袭和转移的影响及作用机制。方法：采用实时荧光定量PCR(quantitative real-time polymerase chain reaction， qRT-PCR)和免疫组织化学检测TRIM14在80例子宫内膜癌组织和30例正常子宫内膜组织中的表达水平，χ2检验分析其表达与临床病理特征的关系，Kaplan-Meier生存曲线分析其表达对患者预后的影响。qRT-PCR检测TRIM14 mRNA在子宫内膜癌细胞系和人正常子宫内膜细胞系ESC中的表达；Ishikawa经脂质体分别转染TRIM14 siRNA(si-TRIM14)与对照(si-NC)48 h后，采用Boyden实验检测各组细胞侵袭能力，Transwell实验检测各组细胞转移能力，蛋白质印迹法检测各组细胞中波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)的影响。结果：qRT-PCR和免疫组织化学结果显示，TRIM14在子宫内膜癌组织中的表达显著高于正常子宫内膜组织，其高表达与肌层浸润程度、组织学分级、淋巴结转移显著相关(均P<0.05)，子宫内膜癌组织中TRIM14高表达患者生存期较短。TRIM14 mRNA在子宫内膜癌细胞系Ishikawa，ECC-1，RL95-2中的表达均显著高于人正常子宫内膜细胞系ESC，且在Ishikawa细胞中的表达最高(P<0.05)；转染TRIM14 siRNA后，Ishikawa细胞侵袭和转移能力下降，Vimentin，N-cadherin蛋白的表达减少，E-cadherin蛋白的表达增加(均P<0.05)。结论：TRIM14在子宫内膜癌组织和细胞系中均高表达，且其高表达与肌层浸润程度、组织学分级、淋巴结转移及不良预后相关；同时，TRIM14可能通过上皮-间充质转化促进子宫内膜癌细胞侵袭和转移，TRIM14可以作为侵袭和转移子宫内膜癌患者的不良预后分子及治疗子宫内膜癌的潜在靶点。
关键词：
子宫内膜癌；三结构域蛋白14；免疫组织化学；侵袭；转移
Expression of TRIM14 in endometrial cancer and its effect on invasion and metastasis of endometrial cancer cells
CorrespondingAuthor: HE Min Email: 3036420578@qq.com
DOI: 10.3978/j.issn.2095-6959.2019.10.006
Abstract
Objective: To investigate the relationship between the expression of tripartite motif-containing 14 (TRIM14) in endometrial cancer and the clinical features and prognosis of patients with endometrial cancer, and to investigate its effect on invasion and metastasis of endometrial cancer. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to detect the expression of TRIM14 in 80 cases of endometrial carcinoma and 30 cases of normal endometrium. The relationship between expression and clinical case characteristics was analyzed by χ2 test. Kaplan-Meier survival curve analysis was performed on prognosis. qRT-PCR was used to detect the expression of TRIM14 mRNA in endometrial cancer cell lines and human normal endometrial cell line ESC; Ishikawa was transfected with TRIM14 siRNA (si-TRIM14) and control (si-NC) for 48 h. The Boyden assay was used to detect the invasive ability of each group cells. Transwell assay was used to detect the metastatic ability of each group cells. Western blot was used to detect the effects of Vimentin, N-cadherin and E-cadherin proteins. Results: qRT-PCR and immunohistochemistry showed that the expression of TRIM14 in endometrial cancer tissues was significantly higher than that in normal endometrial tissues. The high expression was significantly correlated with the degree of myometrial invasion, histological grade, and lymph node metastasis (P<0.05), Patients with high expression of TRIM14 have a shorter survival period. The expression of TRIM14 mRNA in endometrial cancer cell lines Ishikawa, ECC-1, RL95-2 was significantly higher than that in human normal endometrial cell line ESC, and the expression of TRIM14 was the highest in Ishikawa cells (P<0.05). the invasiveness and metastasis ability of Ishikawa cells and the expression of Vimentin, N-cadherin protein was decreased after transfected with TRIM14 siRNA, and the expression of E-cadherin protein increased (P<0.05). Conclusion: TRIM14 is highly expressed in endometrial cancer tissues and cell lines, and high expression is associated with myometrial invasion, histological grade, lymph node metastasis and poor prognosis. TRIM14 may promote cell invasion and metastasis by acting on epithelial-mesenchymal transition. TRIM14 can be used as a poor prognostic molecule for patients with invasive endometrial cancer and a potential target for the treatment of endometrial cancer.
Keywords：
endometrial cancer; tripartite motif-containing 14; immunohistochemistry; invasion; metastasis