摘要
目的:研究miR-4306在骨肉瘤细胞中的表达及其对骨肉瘤细胞生物学行为的影响。方法:选取人成骨细胞hFOB 1.19和骨肉瘤细胞Saos-2、MNNG/HOS CI#5,采用qRT-PCR检测miR-4306表达水平。将miR-4306 mimics和阴性对照分别转染至骨肉瘤细胞Saos-2和MNNG/HOS CI#5,采用CCK-8法和克隆形成实验检测细胞增殖,Transwell实验检测细胞迁移及侵袭,蛋白质免疫印迹检测细胞内上皮-间质转化(epithelial-mesenchymal transition,EMT)标志蛋白表达水平。通过在线靶基因预测网站Targetscan、Starbase预测miR-4306靶基因可能为富含AT序列特异性结合蛋白2(special AT-rich sequence-binding protein 2,SATB2),荧光素酶报告系统鉴定靶向关系。将miR-4306 mimics、LV-SATB2共转染至骨肉瘤细胞中,检测细胞增殖、迁移、侵袭以及EMT标志蛋白表达水平变化。结果:与人成骨细胞hFOB 1.19比较,骨肉瘤细胞Saos-2、MNNG/HOS CI#5中miR-4306表达明显降低(P均<0.01)。转染miR-4306 mimics后,骨肉瘤细胞增殖、迁移、侵袭能力下降,N-钙黏蛋白、血管内皮生长因子表达水平显著降低,E-钙黏蛋白表达水平显著升高(P<0.05或<0.01)。SATB2是miR-4306下游的直接靶标;与miR-4306 mimics+LV-NC组比较,miR-4306 mimics+LV-SATB2组部分逆转miR-4306对骨肉瘤细胞增殖、侵袭、迁移和EMT的影响。结论:miR-4306在骨肉瘤细胞中呈低表达,其可通过下调SATB2表达抑制骨肉瘤细胞增殖、迁移、侵袭和EMT。
Objective:To study the expression of miR-4306 in osteosarcoma cells and its effect on the biological behavior of osteosarcoma cells.Methods:Human osteoblast hFOB 1.19 and osteosarcoma cells Saos-2,MNNG/HOS CI#5 were selected,and qRT-PCR was used to detect the expression level of miR-4306.miR-4306 mimics and negative control were transfected into osteosarcoma cells Saos-2 and MNNG/HOS CI#5,respectively;and cell proliferation was detected by CCK-8 assay and colony formation assay,cell migration and invasion were detected by Transwell assay,and Western blotting was used to detect the expression levels of epithelial-mesenchymal transition(EMT)marker proteins.The online target gene prediction websites Targetscan and Starbase predicted that the target gene of miR-4306 might be special AT-rich sequence-binding protein 2(SATB2),and the luciferase reporter system was used to identify the target relationship.miR-4306 mimics and LV-SATB2 were co-transfected into osteosarcoma cells,and the changes in cell proliferation,migration,invasion and EMT marker protein expression levels were detected.Results:Compared with human osteoblast hFOB 1.19,the expression of miR-4306 in osteosarcoma cells Saos-2 and MNNG/HOS CI#5 was significantly decreased(both P<0.01).After transfection with miR-4306 mimics,the proliferation,migration and invasion abilities of osteosarcoma cells were significantly decreased,the expression levels of N-cadherin and vascular endothelial growth factor were significantly decreased,and the expression level of E-cadherin was greatly increased(P<0.05 or<0.01).SATB2 is a direct downstream target of miR-4306;compared with the miR-4306 mimics+LV-NC group,the miR-4306 mimics+LV-SATB2 group partially reversed the effects of miR-4306 on osteosarcoma cell proliferation,invasion,migration and EMT.Conclusion:miR-4306 is lowly expressed in osteosarcoma cells,and it could inhibit osteosarcoma cell proliferation,migration,invasion and EMT by down-regulating the expression of SATB2.
作者
李书林
廖艺琳
周子杰
肖郑伟
苏凌波
李俊杰
蓝青
陈名迪
郭徽灵
颜来鹏
汤发强
LI Shulin;LIAO Yilin;ZHOU Zijie;XIAO Zhengwei;SU Lingbo;LI Junjie;LAN Qing;CHEN Mingdi;GUO Huiling;YAN Laipeng;TANG Faqiang(Provincial Clinical Medical College,Fujian Medical University,Fuzhou Fujian 350001;The First Department of Orthopedics,Fujian Provincial Hospital,Fuzhou Fujian 350001;College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou Fujian 350001;Integrated Chinese and Western Medicine College,Fujian University of Traditional Chinese Medicine,Fuzhou Fujian 350001,China)
出处
《江苏大学学报（医学版）》
CAS
2023年第1期42-48,共7页
Journal of Jiangsu University：Medicine Edition
基金
福建省卫生计生委医学创新课题(2018-CXR-2)。
