miR-194-3p靶向调控MMP-11抑制骨肉瘤细胞的增殖、迁移和侵袭

目的:研究miR-194-3p对骨肉瘤细胞增殖、迁移和侵袭的影响，并探讨其机制。方法:实验设置空白对照组（control）、miR-194-3p-mimics组（转染 miR-194-3p-mimics）、miR-194-3p-inhibitor组（转染 miR-194-3p-inhibitor）、siRNA-MMP-11组（转染 siRNA-MMP-11）、ovRNA-MMP-11组（转染ovRNA-MMP-11）、miR-194-3p-mimics+ovRNA-MMP-11组（转染miR-194-3p-mimics+ovRNA-MMP-11）,均以脂质体法转染至人成骨肉瘤细胞Saos-2；MTT法检测细胞增殖率；Transwell实验检测细胞侵袭能力；细胞划痕实验检测细胞迁移能力;蛋白印迹实验（Western blot）检测E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、基质金属蛋白酶 11（MMP-11）蛋白表达；实时荧光定量PCR（qRT-PCR）检测miR-194-3p的表达水平。结果:MTT法检测结果显示:miR-194-3p-mimics组、siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组会降低细胞增殖，并与空白对照组和ovRNA-MMP-11组相比有统计学差异（P＜0.05）。细胞划痕实验检测结果显示:miR-194-3p-mimics组、siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组细胞愈合率明显低于ovRNA-MMP-11组（P＜0.05）。细胞侵袭实验检测结果显示:siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组降低细胞侵袭能力，并与空白对照组和ovRNA-MMP-11组相比有统计学差异（P＜0.05）。qRT-PCR检测结果显示:miR-194-3p-mimics组中miR-194-3p的表达水平与空白对照组、miR-194-3p-inhibitor组、siRNA-MMP-11组、ovRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组相比均有统计学差异（P＜0.05）。结论:miR-194-3p在人成骨肉瘤细胞中高表达，并可抑制骨肉瘤细胞的增殖、迁移和侵袭，其机制可能与靶向调控MMP-11所参与的上皮间充质转化（epithelial mesenchymal transition，EMT）信号通道有关，miR-194-3p将来可能为骨肉瘤的预防和治疗提供新靶点。

MMP-11 by targeted miR-194-3p inhibits the proliferation,migration and invasion of osteosarcoma cells

Objective:To study the effects of miR-194-3p on proliferation，migration and invasion of osteosarcoma cells and explore its mechanism．Methods:The blank control group，miR-194-3p-mimics group （transfected miR-194-3p-mimics），miR-194-3p-inhibitor group （transfected miR-194-3p-inhibitor），siRNA-MMP-11 group （transfected siRNA-MMP-11），ovRNA-MMP-11 group （transfected ovRNA-MMP-11），miR-194-3p-mimics+ovRNA-MMP-11 group （transfected miR-194-3p-mimics+ovRNA-MMP-11），all were transfected to Saos-2 cells by liposome method.MTT assay was used to detect the cell proliferation rate.Cell migration was measured by Transwell chamber.Scratch assay was used to detect the cell migration.The expressions of E-cadherin,N-cadherin and MMP-11 were detected by Western blot.qRT-PCR was used to detect the expression of miR-194-3p.Results:MTT assay showed that miR-194-3p-mimics group,siRNA-MMP-11 group,miR-194-3p-mimics+ovRNA-MMP-11 group could reduce cell proliferation,which was statistically significant compared with the control group and ovRNA-MMP-11 group (P＜0.05).The results of cell scratch showed that the healing rate of miR-194-3p-mimics group,siRNA-MMP-11 group,miR-194-3p-mimics+ovRNA-MMP-11 group was significantly lower than that of ovRNA-MMP-11 group （P＜0.05).Transwell test showed that siRNA-MMP-11 group,miR-194-3p-mimics+ovRNA-MMP-11 group decreased the ability of cell invasion,which was statistically significant compared with the control group and ovRNA-MMP-11 group (P＜0.05).qRT-PCR showed that there were significant differences in miR-194-3p expression between miR-194-3p-mimics group and control group,miR-194-3p-inhibitor group,siRNA-MMP-11 group,ovRNA-MMP-11 group,miR-194-3p-mimics+ovRNA-MMP-11 group (P＜0.05).Conclusion:miR-194-3p is highly expressed in human osteosarcoma cells,which can inhibit the proliferation,invasion and metastasis of osteosarcoma cells.The above effects of miR-194-3p may be realized by targeting the epithelial mesenchymal transition (EMT) signal channel involved in MMP-11.miR-194-3p may provide new targets for the prevention and treatment of osteosarcoma in the future.