miR-199a-5p对宫颈癌CaSki细胞放射敏感性的影响及机制研究  

The effect and mechanism of miR-199a-5p on the radiosensitivity of cervical cancer CaSki cells

作　　者：王光霞[1] 董立新[1] 曹丽艳[1] Wang Guangxia;Dong Lixin;Cao Liyan(Department of Oncology,the First Hospital of Qinhuangdao,Qinhuangdao 066000,China)

机构地区：[1]河北省秦皇岛市第一医院肿瘤科,066000

出　　处：《中国医师杂志》2021年第9期1366-1370,共5页Journal of Chinese Physician

摘　　要：目的探讨miR-199a-5p对宫颈癌CaSki细胞放射敏感性的影响及潜在机制。方法体外培养宫颈癌CaSki细胞。采用脂质体Lipofectamine 2000将miR-199a-5p模拟物(miR-199a-5p mimics)转染至宫颈癌CaSki细胞,记为miR-199a-5p组,以转染模拟物对照(mimics control)的CaSki细胞为阴性对照(NC组),以未转染的CaSki细胞为空白对照(Control组),各组细胞经X射线照射后,采用实时荧光定量PCR(qRT-PCR)检测各组细胞中miR-199a-5p的表达量,通过克隆形成实验和流式细胞凋亡实验检测miR-199a-5p对CaSki细胞放射敏感性和凋亡的影响,使用生物信息学软件预测miR-199a-5p的靶基因,通过双荧光素酶报告基因实验和蛋白免疫印迹(Western blot)验证miR-199a-5p和甲状腺激素受体相互作用因子4(TRIP4)的靶向关系。结果转染miR-199a-5p mimics后,miR-199a-5p组中miR-199a-5p的表达量显著高于Control组(P<0.05),经X射线照射后,miR-199a-5p过表达更明显(P<0.05);过表达miR-199a-5p降低CaSki细胞克隆形成能力(P<0.05),促进CaSki细胞的凋亡;且过表达miR-199a-5p能够进一步降低放射后细胞的克隆形成,并促进放射后细胞凋亡(P<0.05);双荧光素酶报告基因实验和Western blot证实了miR-199a-5p能够靶向负调控TRIP4。结论miR-199a-5p可通过靶向负调控TRIP4的表达提高宫颈癌CaSki细胞的放射敏感性。Objective To investigate the effect and potential mechanism of miR-199a-5p on the radiosensitivity of cervical cancer CaSki cells.Methods Cervical cancer CaSki cells were cultured in vitro.MiR-199a-5p mimics(miR-199a-5p mimics)were transfected into cervical cancer CaSki cells(miR-199a-5p group)with liposome by Lipofectamine 2000.CaSki cells transfected with mimics control were used as negative control(NC group)and non transfected CaSki cells were used as blank control(control group).After X-ray irradiation,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-199a-5p in cells of each group.The effects of miR-199a-5p on radiosensitivity and apoptosis of CaSki cells were detected by clone formation assay and flow cytometry apoptosis assay.Bioinformatics software was used to predict the target gene of miR-199a-5p.Double luciferase reporter gene assay and Western blot were used to verify the targeting relationship between miR-199a-5p and thyroid hormone receptor interaction factor 4(TRIP4).Results The expression of miR-199a-5p in miR-199a-5p group was significantly higher than that in control group(P<0.05).After X-ray irradiation,the expression of miR-199a-5p was more obvious(P<0.05);Overexpression of miR-199a-5p could reduce the clonogenic ability and promote the apoptosis of CaSki cells(P<0.05);Overexpression of miR-199a-5p could further reduce the clonal formation and promote the apoptosis of irradiated cells(P<0.05);Double luciferase reporter gene experiment and Western blot confirmed that miR-199a-5p could target and negatively regulate TRIP4.Conclusions miR-199a-5p can increase the radiosensitivity of cervical cancer CaSki cells by negatively regulating the expression of TRIP4.

关 键 词：宫颈肿瘤 CASKI细胞 miR-199a-5p 放射敏感性 TRIP4 

分 类 号：R737.33[医药卫生—肿瘤]