[an error occurred while processing this directive]
中华放射肿瘤学杂志
   2025年4月7日 星期一     首 页 |  期刊介绍  |  编 委 会  |  投稿指南  |  期刊订阅  |  广告合作  |  学术影响  |  收录情况  |  联系我们  |  English
中华放射肿瘤学杂志  2021, Vol. 30 Issue (9): 949-955    DOI: 10.3760/cma.j.cn113030-20200209-00051
物理·技术·生物 最新目录| 下期目录| 过刊浏览| 高级检索 [an error occurred while processing this directive]|[an error occurred while processing this directive]
Myosin X对肺癌H1975细胞系放射敏感性调节及机制探讨
沈惠, 欧海斌, 邵晶, 江耀飞, 刘羽, 张俊红, 谢丛华
武汉大学中南医院放化疗科 湖北省肿瘤生物学行为重点实验室 湖北省肿瘤医学临床研究中心 湖北省肿瘤放疗质量控制中心 430071
Regulation and mechanism of Myosin X on radiosensitivity of non-small cell lung cancer cell line H1975 in vitro
Shen Hui, Ou Haibin, Shao Jin, Jiang Yaofei, Liu Yu, Zhang Junhong, Xie Conghua
Department of Radiation and Medical Oncology, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Cancer Clinical Study Center, Hubei Cancer Radiation Therapy Quality Control Center, Zhongnan Hospital of Wuhan University, Wuhan 430071, China
全文: PDF (0 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      背景资料
摘要 目的 探讨Myosin X对肺癌H1975细胞系放射敏感性调节及相关机制。方法 蛋白免疫印迹法检测肺癌细胞系中Myosin X表达量以获取实验对象。运用CRISPR/Cas9技术建立敲除Myosin X的H1975细胞系(KO组)和感染对照病毒的H1975细胞系(NC组),并进行敲除效率验证。克隆形成实验及多靶单击模型检测两组细胞对射线敏感性差异。γ-H2AX焦点形成实验及RNAseq差异分析技术寻找可能参与Myonsin X介导的肺癌细胞系放射敏感性调节机制。结果 Myosin X在H1975细胞中表达量明显高于其他细胞系(P<0.01)。经鉴定成功构建了Myosin X sgRNA-Lenti-CRISPR v2慢病毒载体,嘌呤霉素筛选后得到Myosin X完全敲除的H1975(KO组)及对照组(NC组)细胞系。克隆形成实验证明相较于NC组,KO组对射线的敏感性更强 (D0值从1.28Gy下降至1.03Gy,SF2从0.29下降至0.21,放射敏感性比为1.24)。γ-H2AX焦点形成实验显示照后1、6h KO组形成的损伤焦点数均多于NC组(P<0.05),RNAseq差异分析技术结果显示KO组ISLR蛋白表达明显低于NC组(P<0.05)。结论 敲除Myosin X可以增强肺癌H1975细胞的放射敏感性,其机制可能与干扰DNA双链损伤修复以及降低ISLR表达有关。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
沈惠
欧海斌
邵晶
江耀飞
刘羽
张俊红
谢丛华
关键词 肌球蛋白XCRISPR/Cas9技术放射敏感性H1975细胞系    
AbstractObjective To investigate the effect and mechanism of Myosin X on the radiosensitivity of non-small cell lung cancer (NSCLC) cell line H1975 in vitro. Methods Western blot was applied to detect the expression level of Myosin X expression. The H1975 cell line with stable knockout of Myosin X (KO group) and infected with control virus (NC group) were constucted by using CRISPR/Cas9 technique. The knockout efficiency was validated. The radiosensitivity of two cell lines was measured by colony formation assay and single-hit multi-target model. γ-H2AX focus formation test and RNA sequencing (RNAseq) analysis were employed to identify the regulatory mechanism of the radiosensitivity of lung cancer cell lines mediated by Myosin X. Results The expression level of Myosin X in the H1975 cells was significantly up-regulated than those in other NSCLC cell lines (all P<0.01). The lentiviral vector of Myosin X sgRNA-Lenti-CRISPR v2 was successfully constructed. After the puromycin screening, H1975 cell lines with complete knockout of Myosin X and control cell lines (NC group) were obtained. Colony formation assay demonstrated that compared with the NC group, the radiosensitivity in the KO group was significantly higher (The D0 value was decreased from 1.28Gy to 1.03Gy, SF2decreased from 0.29 to 0.21, and the sensitization ratio was 1.24). The γ-H2AX focus formation test showed that the number of damage focus formed at 1h and 6h after irradiation in the KO group was significantly larger than that in the NC group (P<0.05. RNAseq analysis indicated that the expression level of ISLR in the KO group was significantly down-regulated than that IN the NC group (P<0.05). Conclusion Knockout of Myosin X can increase the radiosensitivity of H1975 cells probably by interfering the repair of DNA double-strand damage and down-regulating the expression level of ISLR.
Key wordsMyosin X    CRISPR/Cas9 technique    Radiosensitivity    H1975 cell line   
收稿日期: 2020-02-09     
基金资助:国家自然科学基金项目(81001099、81370070);武汉大学中南医院科技创新培育基金(cxpy2017027);武汉大学中南医院医学科技创新平台支撑项目(PTXM2019030)
通讯作者: 刘羽,Email:liuyu97@whu.edu.cn   
引用本文:   
沈惠,欧海斌,邵晶等. Myosin X对肺癌H1975细胞系放射敏感性调节及机制探讨[J]. 中华放射肿瘤学杂志, 2021, 30(9): 949-955.
Shen Hui,Ou Haibin,Shao Jin et al. Regulation and mechanism of Myosin X on radiosensitivity of non-small cell lung cancer cell line H1975 in vitro[J]. Chinese Journal of Radiation Oncology, 2021, 30(9): 949-955.
链接本文:  
http://journal12.magtechjournal.com/Jweb_fszlx/CN/10.3760/cma.j.cn113030-20200209-00051     或     http://journal12.magtechjournal.com/Jweb_fszlx/CN/Y2021/V30/I9/949
京公网安备11010502022167号
版权所有 © 2010《中华放射肿瘤学杂志》编辑部
地址:北京朝阳区潘学园南里17号 中国医学科学院肿瘤医院(100021)
电话:010-67700737,87788294 Email: cjron@cmaph.org
本系统由北京玛格泰克科技发展有限公司设计开发 技术支持:Support@magtech.com.cn