基础医学与临床 ›› 2021, Vol. 41 ›› Issue (2): 233-239.

• 研究论文 • 上一篇    下一篇

miR-491-5p参与调节人膀胱癌细胞系EJ对5-氟尿嘧啶的敏感性

杨锋, 徐佳, 仇国辉, 杨志玲*   

  1. 湖南省人民医院 药学二部,湖南 长沙 410002
  • 收稿日期:2020-03-11 修回日期:2020-06-04 出版日期:2021-02-05 发布日期:2021-01-19
  • 通讯作者: *2295718962@qq.com

miR-491-5p regulates the sensitivity of human bladder cancer cell line EJ to 5-fluorouracil

YANG Feng, XU Jia, QIU Guo-hui, YANG Zhi-ling*   

  1. No. 2 Department of Pharmacy, Hunan People's Hospital, Changsha 410002, China
  • Received:2020-03-11 Revised:2020-06-04 Online:2021-02-05 Published:2021-01-19
  • Contact: *2295718962@qq.com

摘要: 目的 探讨miR-491-5p对人膀胱癌细胞系EJ 5-氟尿嘧啶(5-fluorouracil,5-FU)敏感性的影响及初步机制。方法 RT-qPCR检测miR-491-5p在膀胱癌化疗敏感与耐药组织中表达水平。构建5-FU耐药的EJ/5-FU细胞系,并分别用RT-qPCR和Western blot检测miR-491-5p表达和AKT与STAT3磷酸化水平。将miR-491-5p抑制剂转入细胞系EJ或将miR-491-5p模拟物转入EJ/5-FU细胞,并在5-FU存在的条件下分别用CCK-8法、Annexin V-FITC/PI染色法、划痕法、Transwell小室法和Western blot检测细胞活性、凋亡、伤口愈合、迁移、侵袭和AKT与STAT3磷酸化水平。结果 miR-491-5p在化疗耐药组织和EJ/5-FU细胞系中表达显著降低(P<0.01)。下调miR-491-5p表达后,5-FU对细胞系EJ的细胞活性、伤口愈合、迁移和侵袭的抑制作用和凋亡的促进作用均显著减弱(P<0.01);上调miR-491-5p表达后,5-FU对EJ/5-FU细胞系的细胞活性、伤口愈合、迁移和侵袭的抑制作用和凋亡的促进作用均显著增加(P<0.01)。AKT及STAT3磷酸化水平在EJ/5-FU细胞系中显著升高(P<0.01);下调miR-491-5p后,细胞系EJ中AKT及STAT3磷酸化水平显著升高(P<0.01);上调miR-491-5p后,EJ/5-FU细胞系中AKT及STAT3磷酸化水平显著降低(P<0.01)。结论 下调miR-491-5p表达降低细胞系EJ对5-FU的敏感性;上调miR-491-5p表达抑制EJ/5-FU细胞系对5-FU耐药性。miR-491-5p调节细胞系EJ 5-FU耐药性的过程中伴随着AKT及STAT3磷酸化的改变。

关键词: miR-491-5p, 膀胱癌, 5-氟尿嘧啶

Abstract: Objective To investigate the role and mechanism of miR-491-5p on 5-fluorouracil (5-FU) sensitivity in human bladder cancer cell line EJ. Methods The expressions of miR-491-5p in chemotherapy-sensitive and resistant tissues were detected by RT-qPCR. The 5-FU resistant EJ/5-FU cell line was constructed, the miR-491-5p expression and phosphorylation of AKT and STAT3 were detected by RT-qPCR and Western blot, respectively. After miR-491-5p inhibitor was transfected into EJ cells or miR-491-5p mimic was transfected into EJ/5-FU cells, the cell viability, apoptosis, wound healing, migration, invasion, AKT and STAT3 phosphorylation levels were detected by CCK-8 assay, Annexin V-FITC/PI staining, scratch assay and Transwell assay and Western blot under the condition of 5-FU, respectively. Results The expression of miR-491-5p was significantly decreased in chemotherapy-resistant tissues and EJ/5-FU cell line(P<0.01). After down-regulation of miR-491-5p in EJ cells, the inhibition of 5-FU on cell viability, wound healing, migration and invasion, and the promotion of apoptosis were significantly reduced (P<0.01). After up-regulation of miR-491-5p in EJ/5-FU cell line, the inhibition of 5-FU on cell viability, wound healing, migration and invasion, and the promotion of apoptosis were significantly increased(P<0.01). The phosphorylation of AKT and STAT3 was significantly increased in EJ/5-FU cell line(P<0.01); the phosphorylation of AKT and STAT3 in cell line EJ was significantly increased after the down-regulation of miR-491-5p (P<0.01), and the phosphorylation of AKT and STAT3 in EJ/5-FU cell line was significantly decreased after the up-regulation of miR-491-5p (P<0.01). Conclusions Down-regulation of miR-491-5p reduces the sensitivity of cell line EJ to 5-FU, while up-regulation of miR-491-5p inhibits 5-FU resistance in EJ/5-FU cell line. miR-491-5p in regulation of 5-FU resistance of cell line EJ is accompanied by changes of AKT and STAT3 phosphorylation.

Key words: miR-491-5p, bladder cancer, 5-fluorouracil

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