miRNA-95在骨肉瘤组织中的表达及其对MG-63细胞增殖和侵袭的影响

目的：研究miRNA-95在骨肉瘤组织和细胞株中的表达情况及其对骨肉瘤MG-63细胞增殖、凋亡、细胞周期和侵袭 能力的影响。方法：以实时荧光定量PCR检测15例骨肉瘤组织及其癌旁组织（标本收集自2015年1月至2018年1月青岛市海 慈医疗集团外科手术的病例）和骨肉瘤细胞株（MG-63、 U2OS、143B和HOS）与正常人成骨细胞株hFOB1.19中的miRNA-95表 达。利用Lipofectamine 2000将miRNA-95 mimics和miRNA-95 inhibitors分别转染至人骨肉瘤MG-63细胞株中，并设置miRNANC对照组，CCK-8法检测各组细胞增殖活力变化，流式细胞术检测各组细胞周期和凋亡变化，Transwell方法检测各组细胞侵袭 能力变化，双荧光素酶活性实验检测并验证miRNA-95在MG-63细胞中的靶向基因。结果：miRNA-95在人骨肉瘤组织中的表 达水平显著高于癌旁组织（P<0.01）， 在MG-63、 U2OS、143B和HOS细胞中的表达水平显著高于hFOB1.19，且在MG-63细胞中表 达水平最高（P<0.01）。与miRNA-NC对照组相比，miRNA-95 mimics组中MG-63细胞的增殖活力显著上升、细胞凋亡率显著下 降而侵袭率显著上升（均P<0.01）、 而miRNA-95 inhibitors组中MG-63细胞增殖活力显著下降、细胞周期被阻滞、细胞凋亡率显著 上升而侵袭率显著下降（均P<0.01）；miRNA-95在骨肉瘤MG-63细胞中靶向上皮膜蛋白-1（epithelial membrane protein-1，EMP-1） 基因发挥作用。结论：miRNA-95在人骨肉瘤组织和细胞中均呈高表达，抑制骨肉瘤MG-63细胞中miRNA-95表达能够促进细 胞凋亡进而抑制细胞增殖、细胞周期及侵袭能力，该作用可能通过靶向EMP-1基因而发挥。

Expression of miRNA-95 in osteosarcoma tissues and its effect on proliferation and invasion of MG-63 cells

Objective: To study the expression of miRNA-95 in osteosarcoma tissues and cell lines, as well as to reveal its effect on proliferation, apoptosis, cell cycle and invasion ability of osteosarcoma cells. Methods: Real-time fluorescent quantitative PCR was used to detect the expression of miRNA-95 in 15 pairs of osteosarcoma tissues and their adjacent normal tissues (specimens were collected from patients underwent surgery in Qingdao Haici Medical Group from January 2015 to January 2018), osteosarcoma cell lines (MG-63, U2OS, 143B and HOS) and normal human osteoblast hFOB1.19 cell line. miRNA-95 mimics and miRNA-95 inhibitors were respectively transfected into MG-63 cells by Lipofectamine 2000, and miRNA-NC group was set up as control group. CCK-8 method was used to detect the changes in cell proliferation, Flow cytometry was used to detect the changes in cell cycle and apoptosis, Transwell method was used to detect the changes in cell invasion ability, and Dual luciferase enzyme activity assay was used to detect and validate the target gene of miRNA-95 in osteosarcoma cells. Results: The expression level of miRNA-95 in human osteosarcoma tissues and cell lines (MG-63, U2OS, 143B and HOS) was significantly higher than that in adjacent tissues and normal human osteoblast hFOB1.19 cell line (all P<0.01), with the highest expression in MG-63 cells (P<0.01). Compared with the miRNA-NC group, the proliferation and invasion abilities of MG-63 cells in miRNA-95 mimics group increased significantly, while the apoptosis rate decreased significantly (all P<0.01). However, the proliferation and invasion activities of MG-63 cells in miRNA-95 inhibitor group decreased significantly, while the apoptosis rate increased significantly, and the cell cycle was obviously blocked (all P<0.01). miRNA-95 played a role in targeting the gene of epithelialmembraneprotein1 (EMP-1) in human osteosarcoma MG-63 cells. Conclusion: miRNA-95 is highly expressed in human osteosarcoma tissues and cells; inhibitor of miRNA-95 expression can promote apoptosis and inhibit proliferation, cell cycle and invasion of osteosarcoma cells, which may be related with targeting EMP-1 gene.