作者：孟圆圆，迟鹏宇，马丽辉，陈敏，孙冬霞，庞岚，李燕

单位：

056000 河北省邯郸市妇幼保健院；通信作者：李燕，E-mail：hdsfyck@163.com

Units：

Handan Maternal and Child Health Hospital，Handan 056000，China；Corresponding Author：LI Yan， E-mail：hdsfyck@163.com

关键词：

子宫内膜癌；微 RNAs；微小 RNA-373；细胞增殖；细胞迁移；大型肿瘤抑制因子 2

Keywords：

Cancer of endometrium；MicroRNAs；MicroRNA-373；Cell proliferation；Cell migration；Largetumor suppressor 2

CLC：

R 737.33

DOI：

DOI：10.3969/j.issn.1008-5971.2020.03.017

Funds：

摘要：

背景　微小 RNA-373（miRNA-373）在多种恶性肿瘤（如肝癌、肺癌）的发生发展过程中具有重要作用，但其在子宫内膜癌（EC）发生发展中的作用尚不完全清楚。目的　探讨 EC 组织 miRNA-373 表达情况及其对EC 细胞增殖、迁移的影响。方法　选取 2012 年 6 月—2013 年 6 月在邯郸市妇幼保健院行手术治疗的 64 例 EC 患者的 EC 组织作为试验组，另选取同期在邯郸市妇幼保健院行子宫切除术的 30 例良性子宫肌瘤患者的正常组织作为对照组。采用实时荧光定量聚合酶链反应（qRT-PCR）检测两组 miRNA-373 表达情况，绘制 Kaplan-Meier 生存曲线以分析不同 miRNA-373 表达情况 EC 患者术后 5 年预后；分别比较转染 miRNA-373 模拟物（miRNA-373 mimic）及其阴性对照物（mimic-NC）、miRNA-373 抑制剂（miRNA-373 inhibitor）及其阴性对照物（inhibitor-NC）的 HEC-1B细胞大型肿瘤抑制基因 2（LATS2）表达情况、转染 96 h 后吸光度值、迁移细胞数量；采用双荧光素酶报告基因系统验证 miRNA-373 与 LATS2 间的靶标关系。结果　（1）试验组 miRNA-373 相对表达量高于对照组（P<0.05）。根据miRNA-373 相对表达量平均值将试验组患者分为低表达组（n=26）和高表达组（n=38），Kaplan-Meier 生存曲线显示，高表达组患者术后 5 年累积生存率为 52.6%，低于低表达组的 84.6%（P<0.05）。（2）转染 miRNA-373 mimic 的HEC-1B 细胞 LATS2 相对表达量低于转染 mimic-NC 的 HEC-1B 细胞（P<0.05）；转染 miRNA-373 inhibitor 的 HEC-1B 细胞 LATS2 相对表达量高于转染 inhibitor-NC 的 HEC-1B 细胞（P<0.05）。转染 96 h 后，转染 miRNA-373 mimic的 HEC-1B 细胞吸光度值高于转染 mimic-NC 的 HEC-1B 细胞（P<0.05）；转染 miRNA-373 inhibitor 的 HEC-1B 细胞吸光度值低于转染 inhibitor-NC 的 HEC-1B 细胞（P<0.05）。转染 miRNA-373 mimic 的 HEC-1B 细胞中迁移细胞数量多于转染 mimic-NC 的 HEC-1B 细胞（P<0.05）；转染 miRNA-373 inhibitor 的 HEC-1B 细胞中迁移细胞数量少于转染inhibitor-NC 的 HEC-1B 细胞（P<0.05）。（3）双荧光素酶报告基因系统检测结果显示，LATS2 含有 miRNA-373 的潜在结合位点。野生型 LATS2 中 miRNA-373 相对荧光强度低于空载对照（P<0.05）；突变型 LATS2 中 miRNA-373相对荧光强度与空载对照比较，差异无统计学意义（P>0.05）。结论　EC 组织 miRNA-373 呈高表达，其可通过直接靶向调节 LATS2 的表达而促进 EC 细胞增殖、迁移，进而影响 EC 患者预后。

Abstract：

Background　It is confirmed that microRNA-373（miRNA-373）plays an important role in theoccurrence and development of kinds of tumors（such as liver cancer and lung cancer），however its role in the occurrenceand development of endometrial cancer（EC）is not yet clear so far. Objective　To analyze the expression of miRNA-373 inEC tissue and its impact on cell proliferation and migration. Methods　EC tissues obtained from 64 patients underwent surgicalresection were selected as experiment group in Handan Maternal and Child Health Hospital from June 2012 to June 2013，meanwhile normal tissues obtained from 30 benign uterine fibroids patients underwent hysterectomy were selected as control group.Real-time fluorescence quantitative polymerase chain reaction（qRT-PCR）was used to detect the expression of miRNA-373 inthe two groups，and Kaplan-Meier survival curve was drawn to analyze the prognosis in EC patients with different expression ofmiRNA-373；comparison of expression of LATS2，optical density 96 hours after transfection and number of migratory cells wasconducted between miRNA-373 mimic and mimic-NC（negative control）transfected HEC-1B cells，between miRNA-373inhibitor and inhibitor-NC（negative control）transfected HEC-1B cells，respectively，moreover dual luciferase reporter genesystem was used to verify the target relationship between miRNA-373 and LATS2. Results　（1）Relative expression quantityof miRNA-373 in experiment group was statistically significantly higher than that in control group（P<0.05）. Of the 64 patientswith EC，26 cases were with low expression of miRNA-373，the other 38 cases were with high expression of miRNA-373；Kaplan-Meier survival curve showed that，the postoperative 5-year cumulative survival rate was 52.6% in EC patients with highexpression of miRNA-373，which was statistically significantly lower than that in EC patients with low expression of miRNA-373of 84.6%（P<0.05）.（2）Relative expression quantity of LATS2 in HEC-1B cells transfected with miRNA-373 mimic wasstatistically significantly lower than that in HEC-1B cells transfected with mimic-NC（P<0.05），while relative expressionquantity of LATS2 in HEC-1B cells transfected with miRNA-373 inhibitor was statistically significantly higher than that inHEC-1B cells transfected with inhibitor-NC（P<0.05）. Optical density in HEC-1B cells transfected with miRNA-373 mimicwas statistically significantly higher than that in HEC-1B cells transfected with mimic-NC 96 hours after transfection，whileoptical density in HEC-1B cells transfected with miRNA-373 inhibitor was statistically significantly lower than that in HEC-1B cells transfected with inhibitor-NC（P<0.05）. Number of migratory cells in HEC-1B cells transfected with miRNA-373mimic was statistically significantly more than that in HEC-1B cells transfected with mimic-NC（P<0.05），while number ofmigratory cells in HEC-1B cells transfected with miRNA-373 inhibitor was statistically significantly less than that in HEC-1Bcells transfected with inhibitor-NC（P<0.05）.（3）Dual luciferase reporter gene system detection results showed that，therewas potential binding site of miRNA-373 in LATS2. Relative intensity of fluorescence of miRNA-373 in wild type of LATS2 wasstatistically significantly lower than that in blank-control（P<0.05），however there was no statistically significant differenceinrelative intensity of fluorescence of miRNA-373 between mutant type of LATS2 and blank-control（P>0.05）. ConclusionExpression of miRNA-373 is high in EC tissue，and it may promote the cell proliferation and migration though direct targetingregulation of LATS2，and then affect the prognosis in patients with EC.

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