摘要
目的建立稳定高表达microRNA-218膀胱癌T24细胞系。方法构建pHBLV-U6-ZsGreen-Puro质粒,制备慢转录病毒载体pHBLV-U6-ZsGreen-Puro-miR-218和pHBLV-U6-ZsGreenPuro-vector转染膀胱癌T24细胞系,用嘌呤霉素筛选并荧光显微镜和qRT-PCR检测。结果转染的细胞在荧光显微镜下呈现出绿色荧光,实时荧光定量PCR(qRT-PCR)显示实验组细胞microRNA-218表达水平比对照组升高250倍。结论该实验成功建立了稳定高表达MicroRNA-218膀胱癌T24细胞系,为进一步研究microRNA-218在膀胱癌中的功能奠定了基础。
Objective To establish stable and high bladder cancer T24 cell line overexpressing microRNA-218 for the further studies on the function of microRNA-218 in bladder cancer. Methods pHBLV-U6-ZsGreen-Puro plasmids and lentivirus of pHBLV-U6-ZsGreen-Puro-vector or pHBLV-U6-ZsGreen-Puro-miR-218 was constructed. Both lentivirus were transfected into bladder cancer T24 cell line respectively. After drug screening with puromycin,the stable cell lines were detected by fluorescence microscope and qRT-PCR. Results Green fluorescence in transfected cell lines was observed by fluorescence microscopy. qRT-PCR showed microRNA-218 expression level of experimental group was 250 folds higher than that of control group. Conclusion The stable and high bladder cancer T24 cell line overexpressing microRNA-218 was established, which provides foundations to further study on the functions of microRNA-218 in bladder cancer.
出处
《中华临床医师杂志（电子版）》
CAS
2014年第15期37-40,共4页
Chinese Journal of Clinicians(Electronic Edition)
基金
国家自然科学基金(81272832)
江苏省自然科学基金(bk2011848)
江苏省六大人才高峰项目(2011-WS-121)
