Wang Shuaihua, Wang Xinlei, Guo Qiang, Wang Guilong, Han Xiangzhen, Li Xiao, Shi Zuo-Wei, He Wen
Department of Pediatric Orthopedics and Bone Oncology, Linyi People's Hospital Affiliated to Shandong University, Linyi, Lanshan, Shandong, P.R. China.
Department of Ophthalmology, Linyi People's Hospital Affiliated to Shandong University, Shandong, P.R. China.
Technol Cancer Res Treat. 2016 Oct;15(5):NP49-59. doi: 10.1177/1533034615601563. Epub 2015 Aug 28.
This study investigated the biological effects of microRNA-126 overexpression in human MG63 osteosarcoma cells. A recombinant plasmid expressing microRNA-126, pcDNA6.2-microRNA-126, was constructed and transfected into MG63 cells. Using real-time fluorogenic quantitative polymerase chain reaction, the microRNA-126 expression was measured in microRNA-126-MG63 group, Ctrl-MG63 group, and blank group. Cell proliferation, cell cycle distribution, cell migration, and invasion were analyzed using methyl thiazolyl tetrazolium assay, flow cytometer, wound-healing assay, and transwell assay, respectively. As expected, microRNA-126 expression was higher in microRNA-126-MG63 group than in Ctrl-MG63 group and blank group (both P < .05). After 48/72 hours of transfection, cell proliferation in microRNA-126-MG63 group was significantly reduced compared to blank group (both P < .05). Compared to blank group, cell population in G0/G1 stage was significantly higher in microRNA-126-MG63 group, accompanied by lower cell numbers in the S and G2/M phases and decreased proliferation index (all P < .05). Wound-healing assay showed a wider scratch width in microRNA-126-MG63 group and reduced cell migration than blank group (both P < .05). Cells overexpressing microRNA-126 exhibited reduced ADAM9 expression levels compared to other 2 groups (all P < .05), suggesting ADAM9 is a target of microRNA-126. Cell proliferation, migration, and invasion rates were reduced in microRNA-126 group after 48/72 hours of transfection, compared with blank group (all P < .05). Cotransfection of pcDNA6.2-microRNA-126 and pMIR-ADAM9 into MG63 cells led to higher cell proliferation, invasion, and migration rates, compared with transfection of pcDNA6.2-microRNA-126 alone (all P < .05). In summary, our data show that microRNA-126 inhibits cell proliferation, migration, and invasion in human osteosarcoma cells by targeting ADAM9.
本研究调查了微小RNA-126过表达对人MG63骨肉瘤细胞的生物学效应。构建了表达微小RNA-126的重组质粒pcDNA6.2-微小RNA-126，并将其转染至MG63细胞中。采用实时荧光定量聚合酶链反应检测微小RNA-126-MG63组、Ctrl-MG63组和空白组中微小RNA-126的表达。分别采用甲基噻唑基四氮唑蓝法、流式细胞仪、划痕愈合实验和Transwell实验分析细胞增殖、细胞周期分布、细胞迁移和侵袭情况。正如预期的那样，微小RNA-126-MG63组中微小RNA-126的表达高于Ctrl-MG63组和空白组（均P < 0.05）。转染48/72小时后，微小RNA-126-MG63组的细胞增殖与空白组相比显著降低（均P < 0.05）。与空白组相比，微小RNA-126-MG63组G0/G1期的细胞数量显著增加，同时S期和G2/M期的细胞数量减少，增殖指数降低（均P < 0.05）。划痕愈合实验显示，微小RNA-126-MG63组的划痕宽度更宽，细胞迁移能力低于空白组（均P < 0.05）。与其他两组相比，过表达微小RNA-126的细胞中ADAM9表达水平降低（均P < 0.05），提示ADAM9是微小RNA-126的一个靶点。转染48/72小时后，微小RNA-126组的细胞增殖、迁移和侵袭率与空白组相比降低（均P < 0.05）。将pcDNA6.2-微小RNA-126和pMIR-ADAM9共转染至MG63细胞中，与单独转染pcDNA6.2-微小RNA-126相比，细胞增殖、侵袭和迁移率更高（均P < 0.05）。综上所述，我们的数据表明微小RNA-126通过靶向ADAM9抑制人骨肉瘤细胞的增殖、迁移和侵袭。
