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

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.