苦参碱通过诱导miR-433-3p表达抑制食管鳞癌放射抵抗  

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

作　　者：刘颖[1] 冯达斌 王绩钊 张丞 张露[4] 张明鑫 陈南征[3] LIU Ying;FENG Dabing;WANG Jizhao;ZHANG Cheng;ZHANG Lu;ZHANG Mingxin;CHEN Nanzheng(Department of Oncology,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Department of Thoracic Surgery,Zhen'an People's Hospital,Shang Luo 711500,China;Department of Thoracic Surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Department of Urology,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Department of Gastroenterology,the First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,China)

机构地区：[1]西安交通大学第一附属医院肿瘤内科,西安710061 [2]镇安县人民医院胸外科,商洛711500 [3]西安交通大学第一附属医院胸外科,西安710061 [4]西安交通大学第一附属医院泌尿外科,西安710061 [5]西安医学院第一附属医院消化内科,西安710077

出　　处：《中国癌症防治杂志》2021年第1期34-39,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：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)。

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q RT-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。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-109 R cells.The Eca-109 and Eca-109 R 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 q RT-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-109 R cells and reestablished the radiosensitivity(P<0.05).The miR-433-3p was lowly expressed in Eca-109 R cells(P<0.05),the knockdown of miR-433-3p enhanced the cell viability of Eca-109 R cells,while the Matrine could relieve the promotion of miR-433-3p knockdown on Eca-109 R cell viability effect(F=5.213,P<0.05).RAD21 was highly expressed in Eca-109 R cells(P<0.05).Overexpression of RAD21 could enhance the cell viability of Eca-109 R cells and reverse the inhibitory effect of miR-433-3p on Eca-109 R 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.

关 键 词：食管磷癌 放射抵抗 苦参碱 miR-433-3p RAD21 

分 类 号：R735.1[医药卫生—肿瘤]