微RNA-455-3p靶向TMED3基因调控宫颈癌细胞凋亡和放射敏感性分子机制研究  

miR-455-3p regulates apoptosis and radiosensitivity of cervical cancer cells by targeting TMED3:the molecular mechanism

作　　者：王朝 葛兴萍 孔霞[2] Wang Zhao;Ge Xingping;Kong Xia(Second Department of Radiotherapy,Yantaishan Hospital,Yantai,Shandong 264000,China;Oncology Department,Shan County Central Hospital,Heze,Shandong 274300,China)

机构地区：[1]烟台山医院放疗二科,山东烟台264000 [2]单县中心医院肿瘤科,山东菏泽274300

出　　处：《中华生物医学工程杂志》2019年第5期610-614,共5页Chinese Journal of Biomedical Engineering

摘　　要：目的探讨微RNA(miR)-455-3p对宫颈癌C33a细胞凋亡和放射敏感性的影响及作用机制。方法不同剂量射线照射C33a细胞,实时荧光定量PCR(qRT-PCR)检测miR-455-3p和跨膜p24运输蛋白3(TMED3)mRNA表达,免疫印迹检测TMED3和Cleaved caspase-3蛋白水平。分别构建miR-455-3p过表达和TMED3沉默的C33a细胞,qRT-PCR检测miR-455-3p和TMED3 mRNA表达,免疫印迹检测TMED3和Cleaved caspase-3蛋白水平,流式细胞仪检测细胞凋亡率,克隆形成实验检测细胞放射敏感性。双荧光素酶报告基因实验验证miR-455-3p和TMED3之间靶向调控关系。结果辐射可降低C33a细胞中miR-455-3p水平(P<0.05),提高TMED3 mRNA和蛋白水平(P<0.05)。miR-455-3p过表达或沉默TMED3可促进C33a细胞凋亡(P<0.05),增强C33a细胞放射敏感性(P<0.05)。miR-455-3p负调控TMED3表达,过表达TMED3降低了miR-455-3p过表达对C33a细胞凋亡的促进作用和放射敏感性的增强作用。结论miR-455-3p可能通过下调TMED3表达促进C33a细胞凋亡,并增强其放射敏感性。Objective To investigate the effect of miR-455-3p on apoptosis and radiosensitivity of cervical cancer C33a cells and its mechanism.Methods C33a cells were irradiated with different radiation doses.The expression levels of miR-455-3p and TMED3 mRNA were detected by qRT-PCR,and the protein levels of TMED3 and Cleaved caspase-3 were detected by Western blotting.The cell apoptosis was detected by flow cytometry,and cell radiosensitivity by clone formation assay.Dual luciferase reporter gene assay was used to validate the targeted regulatory relationship between miR-455-3p and TMED3.Results Radiation decreased the level of miR-455-3p in C33a cells(P<0.05)and increased TMED3 mRNA and protein levels(P<0.05).Overexpression of miR-455-3 or silencing of TMED3 promoted apoptosis(P<0.05)and enhanced radiosensitivity of C33a cells(P<0.05).miR-455-3p negatively regulated the expression of TMED3.Overexpression of TMED3 reduced the promotion of apoptosis and the radiosensitivity enhancement of C33a cells by overexpression of miR-455-3p.Conclusion miR-455-3p may promote apoptosis of C33a cells and enhance its radiosensitivity by down-regulating the expression of TMED3.

关 键 词：宫颈癌 微RNA-455-3p 跨膜p24运输蛋白3 细胞凋亡 放射敏感性 

分 类 号：R737[医药卫生—肿瘤]