高表达微小RNA145前列腺癌细胞系PC-3的建立

The establishment of a stable PC-3 cell line overexpressing microRNA-145

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.