RNA干扰靶向沉默COX-2基因对MG-63骨肉瘤细胞生物学特性的研究  

Effect of siRNA Inhibited COX-2 Gene on Biological Characteristics of MG-63 Cells

作　　者：陈秀锦[1] 杨明宣 崔兆辉[1] 胡旭昌[1] 汪静[1] 安丽萍[1] 马靖琳[1] 王栓科[1] 

机构地区：[1]兰州大学第二临床医学院骨科/甘肃省骨关节疾病研究重点实验室,兰州730000

出　　处：《世界科技研究与发展》2014年第6期668-673,共6页World Sci-Tech R&D

基　　金：甘肃省自然科学基金(1010RJZA1240)资助

摘　　要：目的通过构建小干扰RNA(small interfering RNA,siRNA)降低MG-63骨肉瘤细胞环氧合酶-2(COX-2)基因的表达,并进一步研究其对MG-63骨肉瘤细胞增值、侵袭、迁移能力的影响及分子机制。方法设计靶向干扰COX-2基因的siRNA,通过脂质体转染MG-63骨肉瘤细胞,使其抑制MG-63骨肉瘤细胞COX-2基因的表达,后采用噻唑蓝(MTT)比色法、Transwell小室实验研究其对MG-63骨肉瘤细胞增殖、侵袭、迁移能力的影响,采用RFQ-PCR和Western blot分别从基因和蛋白的水平检测MG-63骨肉瘤细胞侵袭性相关因子基质金属酶(MMP-9)的表达及血管内皮生长因子(VEGF)的表达。结果转染MG-63骨肉瘤细胞后,实验组与阴性对照组和空白对照组比较,通过RFQ-PCR和Western blot检测COX-2基因表达降低约90%(P<0.05),MTT检测MG-63骨肉瘤细胞增值能力明显受到抑制(P<0.05),Transwell实验检测MG-63骨肉瘤细胞侵袭、迁移能力明显下降(P<0.05),经RFQ-PCR、Western blot检测侵袭性相关因子MMP-9和血管内皮生长因子VEGF的mRNA及蛋白表达降低(P<0.05)。空白对照组和阴性对照组比较无明显变化,差异无统计学意义(P<0.05)。结论人MG-63骨肉瘤细胞COX-2基因被抑制后,MG-63骨肉瘤细胞增值、侵袭、迁移能力明显下降。Objective Through establishing the small interfering RNA （siRNA）to decrease the expression of MG-63 osteo-sarcoma cell＇s cyclooxygenase-2 （COX-2）gene,and further research the influence and molecular mechanism of the power for proliferation,invasion and migration for MG-63 osteosarcoma cell.Methods Designed the targeted interference COX-2 gene＇s siRNA,transfected MG-63 osteosarcoma cell by the lipidosome,so as to suppress the expression of COX-2 gene in MG-63 osteosarcoma cell,then we used the MTT colorimetric method,and Transwell small room experiment to research the influence to the power of proliferation,invasion and migration for MG-63 osteosarcoma cell.Using RFQ-PCR and Western blot to detect the expression of the MG-63 osteosarcoma cell invasive related factor matrix metal enzyme（MMP-9）expres-sion and the expression of vascular endothelial growth factor（VEGF）from the gene level and the protein level.Results Af-ter transfecting the MG-63 osteosarcoma cells,comparing the experiment group with the negative control group and the blank control group,through the RFQ-PCR and Western blot detecting COX-2 gene＇s expression about 90%（P〈0. 05 ）,MTT de-tecting the MG-63 osteosarcoma cells＇multiplication capacity which is suppressed obviously（P〈0. 05 ）,using Transwell ex-periment to detect MG-63 osteosarcoma cells＇invasion,migration ability decrease significantly（P〈0. 05 ）,through the RFQ-PCR,Western blot detecting the invasive related factor MMP-9 and vascular endothelial growth factor VEGF＇s mRNA and protein expression reduced （P〈0. 05 ）.The blank control group and the negative control group had no significant change, there was no statistically significant difference（P〈0. 05 ）.Conclusion COX-2 siRNA can significant suppress the COX-2 gene＇s expression in people＇s MG-63 osteosarcoma cell,and then further induce MG-63 osteosarcoma cell＇s power of prolifer-ation,invasion and migration.

关 键 词：MG-63骨肉瘤细胞 RNA干扰 迁移 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]