circ_0001955靶向miR-149对前列腺癌DU145细胞系放射敏感性的实验研究  

Effect of circ_0001955 targeted regulation of miR-149 on the radiosensitivity of prostate cancer DU145 cells

作　　者：李征[1] 张天标[2] 程双蕾 吴小源[3] Li Zheng;Zhang Tianlbiao;Cheng Shuanglei;Wu Xiaoyuan(Department of Urology,Nanyang Central Hospital,Nanyang 473000,China;Department of Urology,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Radiation Oncology,Henan Cancer Hospital,Zhengzhou 450000,China)

机构地区：[1]南阳市中心医院泌尿外科,473000 [2]郑州大学第一附属医院泌尿外科,450000 [3]河南省肿瘤医院放疗科,郑州450000

出　　处：《中华放射肿瘤学杂志》2021年第9期961-967,共7页Chinese Journal of Radiation Oncology

摘　　要：目的探讨circ0001955对前列腺癌DU145细胞系放射敏感性的影响及分子机制。方法将si-con、si-circ0001955、miR-con、miR-149转染至DU145细胞中,分别记为si-con、si-circ0001955、miR-con、miR-149组;将miR-149与pc-circ0001955共转染至DU145细胞记为miR-149+pc-circ0001955组,以未经任何处理的细胞作为空白对照组。实时荧光定量PCR检测circ0001955和miR-149表达水平;MTT检测细胞活力;流式细胞仪检测细胞凋亡;Transwell检测细胞迁移和侵袭;蛋白质印迹法检测MMP-2、MMP-9、Cleaved caspase-3、Cleaved caspase-9、γ-H2AX蛋白表达;细胞克隆形成实验检测细胞放射敏感性;双荧光素酶报告实验验证circ0001955和miR-149的靶向关系。结果细胞中circ0001955高表达,miR-149低表达。沉默circ0001955或过表达miR-149后细胞活性、迁移和侵袭数降低,MMP-2、MMP-9表达水平降低,Cleaved caspase-3、Cleaved caspase-9表达水平升高,细胞凋亡率升高(P<0.05)。4 Gy X线照射后细胞中γ-H2AX表达水平升高,细胞存活分数降低,敏感性比1.38。circ0001955靶向调控miR-149,同时过表达circ0001955和miR-149后细胞增殖活性、迁移和侵袭细胞数升高,细胞凋亡率、细胞存活分数升高,敏感性比0.72。结论沉默circ0001955靶向上调miR-149抑制DU145细胞增殖、迁移、侵袭,诱导细胞周期阻滞,诱导细胞凋亡和增加细胞放射敏感性。Objective To investigate the effect and molecular mechanism of circ_0001955 on the radiosensitivity of prostate cancer DU145 cells.Methods The si-con,si-circ_0001955,miR-con and miR-149 were transfected into DU145 cells and recorded as the si-con group,si-circ_0001955 group,miR-con group,miR-149 group.The miR-149 and pc-circ_0001955 were co-transfected into DU145 cells and recorded as the miR-149+pc-circ_0001955 group.Untreated cells were used as the blank control(NC)group.Real-time quantitative PCR was employed to detect the expression levels of circ_0001955 and miR-149.MTT assay was performed to detect cell viability.Flow cytometry was carried out to detect cell apoptosis.Transwell chamber assay was conducted to observe cell migration and invasion.Western blot was performed to detect the expression levels of MMP-2,MMP-9,Cleaved caspase-3,Cleaved caspase-9 andγ-H2AX proteins.Colony formation assay was employed to determine the cell radiosensitivity.Dual-luciferase reporter assay was conducted to verify the targeting relationship between circ_0001955 and miR-149.Results The circ_0001955 was highly expressed,whereas the miR-149 was lowly expressed in prostate cancer DU145 cells.Silencing circ_0001955 or over-expressing miR-149 could decrease the cell viability,migration and invasion,down-regulate the expression levels of MMP-2 and MMP-9,up-regulate the expression levels of Cleaved caspase-3 and Cleaved caspase-9,and increase the apoptosis rate(all P<0.05).After 4 Gy dose irradiation,the expression level ofγ-H2AX was up-regulated,the cell survival fraction was decreased,and the sensitivity ratio was 1.38.circ_0001955 could targetedly regulate the expression level of miR-149.After simultaneous overexpression of circ_0001955 and miR-149,cell proliferation activity and the number of migrating and invading cells were increased,cell apoptosis rate was decreased,and cell survival fraction was increased,and the sensitivity ratio was calculated as 0.72.Conclusion Silencing circ_0001955 can targetedly up-regulate the expr

关 键 词：circ_0001955 miR-149 放射敏感性 

分 类 号：R737.25[医药卫生—肿瘤]