Runx2真核表达载体的构建及其对骨肉瘤细胞增殖的影响  

Construction of RunX2 Expression Plasmid and its Promotive Role on Osteosarcoma Cell Proliferation

作　　者：代建宁[1] 邓彩霞 景调平 姥伟[1] 杨海旭 DAI Jian-Ning;DENG Cai-Xia;JING Diao-Ping;LAO Wei;YANG Hai-Xu(Department of Orthopedics,the 5th Hospital of PLA, Yinchuan 750004, China;Department of Hyperbaric Oxygen,the 5th Hospital of PLA, Yinchuan 750004, China;Department of Anesthesiology,the 5th Hospital of PLA, Yinchuan 750004, China)

机构地区：[1]解放军第五医院骨科中心,宁夏银川750004 [2]解放军第五医院高压氧,宁夏银川750004 [3]解放军第五医院麻醉手术科,宁夏银川750004

出　　处：《生物技术通讯》2017年第5期643-646,共4页Letters in Biotechnology

摘　　要：目的:构建人源Runx2过表达重组载体,探讨Runx2对骨肉瘤细胞增殖能力的影响。方法:以HEK293细胞总RNA为模板,反转录为c DNA,通过PCR技术扩增Runx2全长基因并连接至真核表达质粒pcDNA3.1,筛选阳性克隆进行序列测定后,转染至骨肉瘤细胞Saos-2和U2-OS,通过MTT实验验证Runx2调节细胞增殖的能力。结果:构建了pcDNA3.1-Runx2真核表达载体,并在骨肉瘤细胞Saos-2和U2-OS中高表达Runx2蛋白;Runx2高表达后,骨肉瘤细胞增殖能力显著增加。结论:构建了Runx2基因高效真核表达载体,并证明Runx2可以促进骨肉瘤细胞的增殖。Objective:To investigate the effect of Runx2on the osteosarcoma cells proliferation.Methods:Thetotal RNA was extracted from HEK293cells,from which the Runx2full length cDNA was amplified through RTPCR,and it was cloned into pcDNA3.1vector.The positive plasmid was selected and transfected intoosteosarcoma cells Saos-2and U2-OS,and biological function of Runx2was examined through MTT assay.Results:We constructed Runx2expression plasmid,and confirmed the highly expression of Runx2aftertransfection of pcDNA3.1-Runx2in Saos-2and U2-OS cells.Runx2over-expression induced cell proliferation.Conclusion:Runx2can accelerate osteosarcoma cell proliferation.

关 键 词：RUNX2 真核表达载体 骨肉瘤细胞 细胞增殖 

分 类 号：Q78[生物学—分子生物学] Q25