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中国癌症防治杂志

中国癌症防治杂志 ›› 2021, Vol. 13 ›› Issue (1): 34-39.doi: 10.3969/j.issn.1674-5671.2021.01.06

• 消化道肿瘤专栏 • 上一篇    下一篇

苦参碱通过诱导miR-433-3p表达抑制食管鳞癌放射抵抗

  

  1. 西安交通大学第一附属医院肿瘤内科；镇安县人民医院胸外科；西安交通大学第一附属医院胸外科；西安交通大学第一附属医院泌尿外科；西安医学院第一附属医院消化内科
  • 出版日期:2021-02-25 发布日期:2021-03-05
  • 通讯作者: 张明鑫 E-mailzmx3115@163.com； 陈南征 E-mail002958@xjtufh.edu.cn
  • 基金资助:
    西安市科技局计划项目［2019114613YX001SF035(1)；2019114613YX001SF034（6）］；陕西省教育厅专项项目（19JK0765）;陕西省自然科学基础研究计划项目（2018JM7090）；西安交通大学第一附属医院青年培育项目（2020QN-32）

Matrine-inhibiting radiation resistance in esophageal squamous cell carcinoma by inducing miR-433-3p expression

  • Online:2021-02-25 Published:2021-03-05

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摘要：  目的 探讨苦参碱在食管鳞癌中对放射抵抗及miR-433-3p表达的影响。方法 构建miR-433-3p及RAD21过表达和敲减放射抵抗Eca-109R细胞，经不同照射剂量（0 Gy、2 Gy、4 Gy、6 Gy及8 Gy）及不同浓度（0.5 mg/mL、1.0 mg/mL、2.0 mg/mL、3.0 mg/mL、4.0 mg/mL和5.0 mg/mL）苦参碱分别处理转染前后的Eca-109和Eca-109R细胞后，采用qRT-PCR及Western blot检测miR-433-3p及RAD21的表达情况，CCK-8实验检测细胞活性；在Eca-109细胞系中，应用双荧光素酶报告基因实验检测miR-433-3p与RAD21的相互作用。结果 不同浓度苦参碱作用可抑制放疗抵抗细胞Eca-109R的细胞活性，重建其对放射的敏感性（P＜0.05）。miR-433-3p在Eca-109R细胞中低表达（P＜0.05）；敲减miR-433-3p可增强Eca-109R细胞活性，而苦参碱能够解除miR-433-3p敲减对Eca-109R细胞活性的促进作用（F=5.213，P＜0.05）。RAD21在Eca-109R细胞中高表达（P＜0.05）；过表达RAD21可增强Eca-109R细胞活性，并逆转miR-433-3p对Eca-109R细胞活性的抑制作用（F=4.554，P＜0.05）。双荧光素酶报告基因提示miR-433-3p可作用于RAD21的3′UTR区域。结论 苦参碱通过诱导miR-433-3p高表达而抑制RAD21表达，重建食管鳞癌的放射敏感性。

关键词: 食管磷癌, 放射抵抗, 苦参碱, miR-433-3p, RAD21

Abstract:  Objective To investigate the effect of Matrine on radiation resistance and miR-433-3p expression in esophageal squamous cell carcinoma. Methods To construct the overexpression of miR-433-3p and RAD21 and knock down the radiation resistance of Eca-109R cells. The Eca-109 and Eca-109R cells before and after transfection were treated with different radiation doses(0 Gy, 2 Gy, 4 Gy, 6 Gy and 8 Gy) and different concentrations(0.5 mg/mL, 1.0 mg/mL, 2.0 mg/mL, 3.0 mg/mL, 4.0 mg/mL and 5.0 mg/mL) of Matrine;  the expression of miR-433-3p and RAD21 were detected by qRT-PCR and western blot, and the cell viability was detected by CCK-8 assay. Dual luciferase reporter gene assay was used to evaluate the interaction between miR-433-3p and RAD21 in Eca-109 cells. Results Different concentrations of Matrine inhibited the cell viability of radiation resistant Eca-109R cells and reestablished the radiosensitivity(P＜0.05). The miR-433-3p was lowly expressed in Eca-109R cells(P<0.05), the knockdown of miR-433-3p enhanced the cell viability of Eca-109R cells, while the Matrine could relieve the promotion of miR-433-3p knockdown on Eca-109R cell viability effect(F=5.213, P＜0.05). RAD21 was highly expressed in Eca-109R cells(P<0.05). Overexpression of RAD21 could enhance the cell viability of Eca-109R cells and reverse the inhibitory effect of miR-433-3p on Eca-109R cell activity(F=4.554, P＜0.05). Dual luciferase reporter gene assay indicated that miR-433-3p specifically interacted with the 3′UTR region of RAD21. Conclusion Matrine can inhibit the expression of RAD21 by inducing the high expression of miR-433-3p and reestablish the radiosensitivity of esophageal squamous cell carcinoma.

Key words:  , Esophageal squamous cell carcinoma, Radiation resistance, Matrine, miR-433-3p, RAD21

中图分类号: 

  • R735.1