摘要
目的:探讨微RNA-29b(microRNA-29b,miR-29b)在骨尤文肉瘤细胞中的表达及其对尤文肉瘤细胞体外侵袭和微血管拟态形成能力的影响。方法:采用实时荧光定量PCR法检测miR-29b在骨尤文肉瘤A673、SKES-1及RD-ES细胞中的表达水平。通过转染外源性miR-29b模拟片段(mimic)使A673和SK-ES-1细胞中miR-29b过表达,然后采用Transwell小室侵袭实验和体外血管拟态形成实验分别检测miR-29b过表达对A673及SK-ES-1细胞侵袭和血管拟态形成能力的影响,并采用蛋白质印迹法检测过表达miR-29b后尤文肉瘤A673和SK-ES-1细胞中信号转导与转录激活因子3(signal transducer and activator factor of transcription 3,STAT3)及其磷酸化STAT3(phosphorylated STAT3,p-STAT3)蛋白表达水平的变化。结果:与人间充质干细胞(对照组)相比,骨尤文肉瘤A673、SK-ES-1和RD-ES细胞中miR-29b表达水平呈下降趋势(P<0.01,P<0.01,P<0.05)。通过转染外源性miR-29b mimic,恢复A673及SK-ES-1细胞中miR-29b的表达后(P值均<0.01),2种细胞的体外侵袭能力均较对照组明显降低(P值均<0.01),体外新生血管形成能力也较对照组明显降低(P<0.05,P<0.01),而且尤文肉瘤细胞中STAT3及p-STAT3的表达水平均明显下调(P值均<0.01)。结论:尤文肉瘤细胞中miR-29b低表达,过表达miR-29b能够明显抑制尤文肉瘤细胞的体外侵袭及新生血管拟态形成,该作用可能与靶向调控STAT3通路有关。
Objective： To investigate the expression of microRNA-29b （miR-29b） in Ewing sarcoma cells, and its effects on the invasion and vascular mimicry formation of Ewing sarcoma cells in vitro.Methods： The expression level of miR-29b in Ewing sarcoma A673, SK-ES-1 and RD-ES cells was detected by real-time fluorescent quantitative PCR. The exogenous miR-29b mimic was transfected into A673 and SK-ES-1 cells to restore the expression of miR-29b in Ewing sarcoma cells. Then the effects of miR-29b overexpression on the invasion and vascular mimicry formation of A673 and SK-ES-1 cells were detected by Transwell chamber invasive assay and in vitro vascular mimicry formation assay, respectively. The expressions of signal transducer and activator factor of transcription 3 （STAT3） and phosphorylated STAT3 （p-STATS） in Ewing sarcoma A673 and SK-ES-1 cells after transfection with miR-29b mimic were detected by Western blotting.Results： Comparing with the human mesenchymal stem cells （as the control group）, the expression level of miR-29b was consistently decreased in Ewing sarcoma A673, SK-ES-1 and RD-ES cells （P 〈 0.01, P 〈 0.01, and P 〈 0.05）. After transfection with exogenous miR- 29b mimic, the expression level of miR-29b was significantly up-regulated in A673 and SK-ES-1 cells （both P 〈 0.01）, the invasive capacity of the two cell lines was significantly suppressed as compared with the control group （both P 〈 0.01）, the in vitro vascular mimicry formation capacity of A673 and SK-ES-1 cells was also significantly suppressed （P 〈 0.05, P 〈 0.01）. Furthermore, the expression levels of STAT3 and p-STAT3 proteins were consistently decreased in Ewing sarcoma A673 and SK-ES-1 cells after transfection with miR-29b mimic （all P 〈 0.01）.Conclusion： miR-29b is lowly expressed in Ewing sarcoma cells. The overexpression of miR- 29b can inhibit the invasion and vascular mimicry formation of Ewing sarcoma ceils in vitro, which may be related to the targeting regulation of STAT3 pathway.
作者
何江孟
陈佳骏
周新
王滕羽
张中卒
HE Jiangmeng;CHEN Jiajunz;ZHOU Xin;WANG Tengyu;ZHANG Zhongzu(Department of Orthopedics, Bishan District Hospital of Traditional Chinese Medicine, Chongqing 402760, China;Department of Orthopedics, Yongchuan Hospital of Chongqing Medical University, Chongqing 402160, China)
出处
《肿瘤》
CAS
CSCD
北大核心
2018年第6期519-525,571,共8页
Tumor
基金
国家自然科学基金资助项目(编号:81502329)~~
