新靶点细胞周期蛋白E干扰RNA对人脑胶质瘤细胞增殖及侵袭的影响

目的探讨新靶点细胞周期蛋白E干扰RNA对人脑胶质瘤细胞的增殖及侵袭转移能力的影响。方法构建4个新靶点的Cyclin E干扰RNA的真核表达载体作为实验组（1至4组），同时设立U251人脑胶质瘤细胞为对照组（U251组）。对实验组用双酶切和碱基序列测定的方法鉴定载体构建成功与否，转染载体到胶质瘤U251细胞株，通过RT-PCR的结果检测转染后Cyclin E mRNA表达量。对实验组及U251组用M1rr法检测细胞增殖能力的变化，用Transwell小室法检测侵袭能力的变化。结果分别构建了4个新靶点的Cyclin E干扰RNA真核表达载体，CyclinEmRNA表达明显受到抑制（P均〈0．05），其中实验1组细胞与U251组相比增殖及侵袭能力减弱（P均〈0．05）。结论新靶点Cyclin E干扰RNA使Cyclin E表达下调，从而导致人脑胶质瘤细胞增殖和侵袭能力减弱。

Effects of Cyclin E RNA interference on the invasive ability and proliferation of U251 human glioma cells

Objective To investigate the effects of Cyclin E RNA interference on theinvasive ability and proliferation of U251 human glioma cells in vitro. Methods Eukaryoticexpression vector of 4 new targets RNA interference specific for Cyclin E were constructed asthe experiment groups （groups 1 to 4） and the U251 human glioma cells group were establishedas control （U251 group）. The experiment groups were tested by restriction enzyme digestion andnucleotide sequence determined and the vectors were transfected into Lipofectamineto U251human glioma cells lines, while the U251 group were untransfected. Then the expression levelsof Cyclin E mRNA were detected by RT-PCR and the invasive ability of U251 human gliomacells was detected by Transwell Migration Assay. Results Eukaryotic expression vectors of newtargets RNA interference specific for Cyclin E were constructed successfully. Cyclin E mRNAexpression in the experiment groups was obviously down-regulated than the U251 group （all P〈0.05）. The proliferative capacity and invasion of U251 human glioma cells in group 1 weresignificantly weakened compared with the U251 group （all P 〈 0.05）. Conclusion Cyclin ERNA interference eukaryotic expression vectors inhibit the expression of Cyclin E, therebyreduce the invasive ability and proliferation of U251 human glioma cells.