天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (2): 155-158.doi: 10.11958/58919

• 细胞与分子生物学 • 上一篇    下一篇

miRNA-7 对食管癌细胞 TE-1 化疗耐药的影响

温爽 1， 杨晓煜 2， 张敏 1， 褚秀峰 1， 钟根深 3， 姬颖华 1△， 路平 1△   

  1. 1新乡医学院第一附属医院肿瘤科(邮编 453100)； 2新乡医学院病理教研室； 3新乡医学院第一附属医院神经病研究所5.
  • 收稿日期:2015-05-11 修回日期:2015-09-19 出版日期:2016-02-15 发布日期:2016-02-15
  • 通讯作者: △通讯作者 E-mail： sunny8441_cn@sina.com； lupingdoctor@126.com E-mail:wenshuang5288@163.com
  • 基金资助:
    新乡医学院科研项目培育基金资助项目 （2013QN127）

The effect of miRNA-7 on chemoresistance in esophageal cancer cell TE-1

WEN Shuang1， YANG Xiaoyu2， ZHANG Min1， CHU Xiufeng1， ZHONG Genshen3， JI Yinghua1△， LU Ping1△   

  1. 1 Department of Oncology, the First Affiliated Hospital of Xinxiang Medical University, Henan 453100, China;2 Department of Pathology, Xinxiang Medical University; 3 Research Institution of Neurology, the First Affiliated Hospital of Xinxiang Medical College
  • Received:2015-05-11 Revised:2015-09-19 Published:2016-02-15 Online:2016-02-15
  • Contact: △Corresponding Author E-mail:sunny8441_cn@sina.com；lupingdoctor@126.com E-mail:wenshuang5288@163.com

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摘要： 目的 探讨微小 RNA-7（miRNA-7）过表达对食管癌细胞 TE-1 顺铂敏感性的影响及其可能机制。方法 用 Lipofectmin 2000 法向食管癌细胞株 TE-1 转染组瞬时转染 miRNA-7 mimic ， 向转染对照组瞬时转染 mimic Negative Control， 通过 RT-PCR 检测以上 2 组及空白对照组的 miRNA-7 以及表皮生长因子受体（EGFR）mRNA 表达情况。Western blot 法分别检测转染组与转染对照组总 EGFR 及细胞浆、 细胞核内的 EGFR 蛋白表达。CCK-8 检测转染组与转染对照组 TE-1 的顺铂半数抑制浓度 （IC50）。免疫荧光共聚焦显微镜观察转染组与转染对照组 EGFR 的表达。结果 转染组较转染对照组及空白对照组的 miRNA-7 表达显著增高， EGFR mRNA 表达下降（均 P＜ 0.001)； 转染组较转染对照组总 EGFR 降低， 核内 EGFR 增高（均 P＜0.01）， 胞浆 EGFR 表达降低（P＜0.05）； CCK-8 结果显示， TE-1 中 miRNA-7 过表达后顺铂（48 h） IC50较转染对照组增高（P＜0.01）； 免疫荧光示转染组较转染对照组核内 EGFR 增高且胞膜与胞浆 EGFR 表达减少。结论 食管癌细胞 TE-1 中 miRNA-7 过表达可通过 EGFR 核转位增多使顺铂产生耐药性。

关键词: 食管肿瘤, 顺铂, 受体, 表皮生长因子, 抗药性, 肿瘤, 微 RNAs, 核转位, 微小 RNA-7

Abstract: Objective To explore the impacts of over-expression of microRNA-7 (miRNA-7) on the sensitivity of cisplatin in esophageal carcinoma cell line TE-1, and the possible mechanism thereof. Methods Lipofectmin 2000 method was used to transient transfect with miRNA-7 mimic into esophageal cancer cell line TE-1, which was taken as transfection group, mimic negative control was taken as transfection conrtol group. The expressions of miRNA- 7 and epidermal growth factor receptor (EGFR) mRNA were detected by RT-PCR in the above two groups and normal control group. The total EGFR and EGFR in cytoplasmic and nucleus were detected with Western blot assay in transfection group and transfection control group. CCK-8 was used to detect IC50 of cisplatin in transfection group and transfection control group. The expression of EGFR was observed with immunofluorescence confocal microscope in two groups. Results The miRNA-7 expression was significantly increased in transfection group than that of transfection conrtol group and control group. The expression of EGFR mRNA was significantly reduced in transfection group (P＜0.001). The total EGFR was significantly decreased in transfection group than that of transfection conrtol group. The level of nuclear EGFR was significantly increased (P＜0.01)， and cytoplasm EGFR expression was significantly decreased in transfection group than that of transfection control group (P＜0.05). CCK-8 results showed that after the over expression of miRNA-7 in TE-1, the IC50 of cisplatin (48 h) increased in transfection group than that of control group (P＜0.01). Immunofluorescence results showed that EGFG in nuclear was higher in transfection group than that of transfection control group but its expressions reduced in cell membrane and cytoplasm. Conclusion The over-expressed miRNA-7 in esophageal cancer cells TE-1 can reduce cisplatin sensitivity by the increased EGFR in nuclear translocation.

Key words: esophageal neoplasms, cisplatin, receptor, epidermal growth factor, drug resistance, neoplasm, microRNAs, nuclear translocation, miRNA-7