当归红芪超滤物对重离子12C6+辐射肝癌细胞DNA损伤修复的影响

辐射防护 ›› 2019, Vol. 39 ›› Issue (4): 331-337.

当归红芪超滤物对重离子¹²C⁶⁺辐射肝癌细胞DNA损伤修复的影响

寇炜¹, 李应东², 窦春江¹, 顾巧玲¹, 刘凯², 陈红³

1.西北民族大学医学院,兰州 730030;
2.甘肃中医药大学中西医结合系,兰州 730000;
3.中国科学研究院近代物理研究所,兰州 730000

收稿日期:2018-11-29 出版日期:2019-07-15 发布日期:2019-10-29
作者简介:寇炜(1972—), 男,1997年毕业于兰州医学院临床医学专业,2003年毕业于兰州大学临床医学院内科学专业,获医学硕士学位,2015年毕业于兰州大学中西医结合基础与临床专业,获医学博士学位,教授。E-mail: yxkw@xbmu.edu.cn
基金资助:
国家自然科学基金项目(81460677);西北民族大学引进人才科研项目资助(xbmuyjrc201629)。

Effects of Angelica sinensis and Radix Hedysari on heavy ion ¹²C⁶⁺irradiation of hepatoma cells DNA damage repair

KOU Wei¹, LI Yingdong², DOU Chunjiang¹, GU Qiaoling¹, LIU Kai², CHEN Hong³

1.The Medical College of Northwest Minzu University,Lanzhou 730030;
2. The Cooperation of Chinese andWestern Medicine Department,Gansu University of Chinese Medicine, Lanzhou 730000;
3. The Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000

Received:2018-11-29 Online:2019-07-15 Published:2019-10-29

摘要/Abstract

摘要： 将人肝癌H22细胞分成4组,分别为对照组、药物组(100 mg/L)、辐射组(2 Gy)及联合组(100 mg/L药物 + 2 Gy照射),采用CCK-8法、单细胞凝胶电泳、γ-H2AX免疫荧光原位杂交技术以及Western Blotting印迹法,研究当归红芪超滤物(Radix Angelicae Sinensis and Radix Hedysari, RAS-RH)对重离子¹²C⁶⁺辐射引起人肝癌H22细胞DNA损伤修复的影响和其可能的机制。结果表明,在0～72 h和给药剂量为5～200 mg/L范围内,RAS-RH对人肝癌H22细胞的增殖抑制作用具有时间和剂量依赖性,其20%抑制浓度IC₂₀为(117.6±2.15)mg/L;单细胞凝胶电泳显示联合组头部DNA含量低于辐射组,而尾部DNA含量、尾距TM、Olive尾距OTM均高于辐射组;γ-H2AX免疫荧光原位杂交技术发现RAS-RH不增加重离子¹²C⁶⁺辐射引起的DNA损伤,但在2—12 h,DNA双链断裂的γ-H2AX foci修复作用被RAS-RH抑制,DNA损伤持续存在;Western Blotting显示RAS-RH通过下调Ku70/80及Rad51的蛋白表达,抑制γ-H2AX的聚集。以上结果说明RAS-RH对人肝癌H22细胞的辐射增敏作用可能是下调DNA损伤修复相关因子Ku70/80及Rad51的表达。

关键词: 当归红芪超滤物, 重离子¹²C⁶⁺, 肝癌H22细胞, DNA损伤修复

Abstract: The paper is mainly about to observe the effect of Radix Angelicae Sinensis and Radix Hedysari (RAS-RH) on hepatoma cells H22 DNA damage induced by heavy ion ¹²C⁶⁺ irradiation, and to explore its possible mechanism. H22 cells were cultured in DMEM medium.They were then divided into control group, drug group(RAS-RH), radiation group(Gy) and combination group(RAS-RH + Gy). CCK-8 method, single cell gel electrophoresis, γ-H2AX immunofluorescence in situ hybridization and Western Blotting were used to study the correlation. The results showed that RAS-RH inhibited the proliferation of human hepatocellular carcinoma H22 cells in a time-and dose-dependent manner at concentrations of 0-72 h and 5-200 mg/L. The IC20 of RAS-RH was (117.6+2.15) mg/L. Single cell gel electrophoresis showed that the content of Head DNA in the combined group was higher than that in the radiation group, while Tail DNA, TM and OTM were lower than those in the radiation group. RAS-RH did not increase DNA damage induced by heavy ion ¹²C⁶⁺ radiation, but in 2-12 h, the repair of double strand breaks of DNA by γ-H2AX foci was inhibited by RAS-RH, and DNA damage persisted. Western Blotting showed that RAS-RH inhibited the aggregation of γ-H2AX by down-regulating the expression of Ku70/80 and Rad51 proteins. It was suggested that RAS-RH could down-regulate the expression of DNA damage repair related factors Ku70/80 and Rad51 in human hepatocellular carcinoma H22 cells.

Key words: RAS-RH, heavy ion ¹²C⁶⁺, hepatoma cells line H22, DNA damage repair

中图分类号:

R142.4

寇炜, 李应东, 窦春江, 顾巧玲, 刘凯, 陈红. 当归红芪超滤物对重离子¹²C⁶⁺辐射肝癌细胞DNA损伤修复的影响[J]. 辐射防护, 2019, 39(4): 331-337.

KOU Wei, LI Yingdong, DOU Chunjiang, GU Qiaoling, LIU Kai, CHEN Hong. Effects of Angelica sinensis and Radix Hedysari on heavy ion ¹²C⁶⁺irradiation of hepatoma cells DNA damage repair[J]. RADIATION PROTECTION, 2019, 39(4): 331-337.

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当归红芪超滤物对重离子¹²C⁶⁺辐射肝癌细胞DNA损伤修复的影响

Effects of Angelica sinensis and Radix Hedysari on heavy ion ¹²C⁶⁺irradiation of hepatoma cells DNA damage repair

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