摘要
目的建立高表达微小RNA145前列腺癌骨转移细胞系,为研究其在骨转移中的作用机制提供依据。方法构建pMSCV-miR-145质粒,制备逆转录病毒pMSCV-vector和pMSCV-miR-145并感染前列腺癌骨转移细胞株PC-3,筛选并鉴定稳定细胞株PC-3/pMSCV-vector与PC-3/pMSCV-miR-145。结果转染pMSCV-miR-145实验组和pMSCV-vector对照组的质粒转染率均为80%~90%。荧光实时定量RT-PCR显示实验组细胞miR-145表达水平与对照组比较差异达2000倍,差异有统计学意义(P<0.05)。表明筛选出的稳定细胞株中,miR-145的表达水平升高。结论高表达微小RNA145前列腺癌细胞系的成功建立,为探索miR-145在前列腺癌骨转移中的作用奠定基础。
Objective To establish stable prostate cancer bone metastasis cell line overexpressing microRNA-145 (miR-145) for the study of the mechanism of miR-145 in bone metastasis. Methods pMSCV-miR-145 plasmids and retroviruses of pMSCV-vector or pMSCV-miR-145 were constructed. Both retroviruses were transfected into bone metastasis PC-3 cell line respectively. After selection and identification, the stable cell lines named PC-3/ pMSCV-vector and PC-3/pMSCV-miR-145 were obtained. Results The proportion of transfected cells were approximately 80%-90% in PC-3/pMSCV-miR-145 cell line group (experimental group) and PC-3/pMSCV-vector cell line group (control group). Real-time fluorescent quantitative RT-PCR showed miR-145 expression level of experimental group was 2 000 folds higher than that of control group, and the difference had statistical signifi-cance between two groups (P〈0.05). Conclusion The establishment of stable prostate cancer bone metastasis cell line overexpressing miR-145 provides foundations to investigate the role of miR-145 in the mechanism of prostate cancer metastasis to bone.
出处
《中国骨科临床与基础研究杂志》
2010年第2期148-152,共5页
Chinese Orthopaedic Journal of Clinical and Basic Research
基金
广东省科技计划项目(2008B030301037)
2009年度珠海市重大科技计划项目
关键词
前列腺肿瘤
骨肿瘤
肿瘤细胞
培养的
Prostatic neoplasms
Bone neoplasms
Tumor cells
cultured