miR-22-3p靶向负调控eIF4E对儿童骨肉瘤细胞增殖影响的研究  

Effect of miR-22-3p targeting negatively regulated eIF4E on the proliferation of osteosarcoma cells in children

作　　者：姚红月 李岩 陈美雪 王金凤 YAO Hongyue;LI Yan;CHEN Meixue(Department of Pediatrics,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000)

机构地区：[1]锦州医科大学附属第一医院儿科,锦州121000 [2]锦州医科大学附属第一医院普外科,锦州121000

出　　处：《陕西医学杂志》2020年第1期16-19,共4页Shaanxi Medical Journal

摘　　要：目的:观察微小RNA-22-3p(miR-22-3p)与真核细胞翻译起始因子4E(eIF4E)的靶向关系,分析其对骨肉瘤细胞增殖的影响。方法:选择骨肉瘤细胞系SAOS-2和成骨细胞系OB-3进行研究,采用双荧光素酶报告基因实验检测miR-22-3p与eIF4E的结合情况。建立空白对照组、空载体转染组、miR-22-3p转染组、miR-22-3p和eIF4E共转染组,采用CCK-8法检测细胞活性,采用Western Blot检测增殖细胞核抗原(PCNA)的表达。应用实时荧光定量PCR法(qPCR)检测miR-22-3p在27例儿童骨肉瘤组织中的表达,应用免疫组化法检测组织中eIF4E和Ki67的表达。结果:骨肉瘤细胞系中miR-22-3p的表达明显低于正常成骨细胞系。双荧光素酶报告基因实验显示miR-22-3p能引起转染pGL3-eIF4E-WT细胞系中荧光素酶活性降低。与空白对照组及空载体转染组相比,miR-22-3p转染组细胞活性下降,PCNA表达下降,miR-22-3p和eIF4E共转染组能逆转上述表达水平。MiR-22-3p在骨肉瘤的不同肿物最大径和病理分级分组的表达中差异有统计学意义(P<0.05)。MiR-22-3p与eIF4E、miR-22-3p与Ki67均具有负相关性,eIF4E与Ki67具有正相关性。结论:miR-22-3p靶向负调控eIF4E调节骨肉瘤细胞的增殖。儿童骨肉瘤中miR-22-3p低表达是肿瘤进展的重要危险因素。Objective:To observe the targeting relationship between microRNA-22-3 p(miR-22-3 p) and eIF4 E,and analyze the effect on the proliferation of osteosarcoma cells. Methods:Osteosarcoma cell line SAOS-2 and osteoblast line OB-3 were selected for research,and the binding of miR-22-3 p to eIF4 E was detected by double luciferase reporter gene assay. The blank control group,empty vector transfection group,miR-22-3 p transfection group,miR-22-3 p and eIF4 E co-transfection group were established. Cell activity was detected by CCK-8 method. The expression of PCNA was detected by Western Blot method. The expression of miR-22-3 p was detected by real-time fluorescent quantitative PCR(qPCR) in 27 children with osteosarcoma,and the expressions of eIF4 E and Ki67 were detected by immunohistochemistry. Results:The expression of miR-22-3 p was significantly lower in osteosarcoma cell line than that in normal osteoblasts cell line. The double luciferase reporter gene experiment showed that miR-22-3 p could reduce the luciferase activity in pGL3-eIF4 E-WT cell line. Compared with the blank control group and the empty vector transfection group,the cell activity and PCNA expression decreased in miR-22-3 p transfection group,miR-22-3 p and eIF4 E co-transfection group could reverse the above expression levels. The expression of miR-22-3 p had statistically significant differences in different tumor maximum diameter and pathological grading of osteosarcoma. Negative correlation was found between miR-22-3 p and eIF4 E,which was also found between miR-22-3 p and Ki67. Positive correlation was found between eIF4 E and Ki67. Conclusion:MiR-22-3 p can negatively regulate eIF4 E to regulate proliferation of osteosarcoma cells. Low expression of miR-22-3 p is an important risk factor for progression of osteosarcoma in children.

关 键 词：骨肉瘤 miR-22-3p 真核细胞翻译起始因子4E 细胞增殖 细胞凋亡 

分 类 号：R738.1[医药卫生—肿瘤]