Shao Ning, Ma Gui, Zhang Jinying, Zhu Wei
Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, China.
Department of Urology, Second People's Hospital of Wuxi, Nanjing Medical University, Wuxi, China.
BMC Urol. 2018 Mar 5;18(1):14. doi: 10.1186/s12894-018-0325-8.
To investigate the effect of miR-221-5p on cell proliferaton and metastasis of human prostate cancer in vitro and vivo.
We established PC3 cell lines with stable overexpression or silencing of miRNA-221-5p via lentivirus infection. miRNA-221-5p and its target gene SOCS1 expression levels in the stable cells were analyzed by real-time polymerase chain reaction (RT-PCR) and western blotting. Using luciferase reporter assays to study the relationship between miR-221-5p and SOCS1. Cell proliferative activity was measured using the MTT assay and colony formation assay. Migration ability was assessed using wound-healing assay and transwell assay. To further study the function of miR-221-5p in human prostate cancer we established nude mice xenograft model in vivo.
miR-221-5p regulates the proliferation, migration of prostate cancer cells in vitro and tumorigenesis in vivo by regulating socs1 expression through targeted its 3'UTR, and miR-221-5p regulates MAPK/ERK signaling pathway and EMT features in prostate cancer cells.
Up-regulation and silencing of miR-221-5p expression in prostate cancer cells are correlated with cell proliferation, migration and tumorigenesis, which suggest that miR-221-5p plays an important role in prostate cancer progression.
研究miR-221-5p在体外和体内对人前列腺癌细胞增殖和转移的影响。
通过慢病毒感染建立miRNA-221-5p稳定过表达或沉默的PC3细胞系。采用实时聚合酶链反应(RT-PCR)和蛋白质免疫印迹法分析稳定细胞中miR-221-5p及其靶基因SOCS1的表达水平。利用荧光素酶报告基因检测法研究miR-221-5p与SOCS1之间的关系。采用MTT法和集落形成试验检测细胞增殖活性。采用伤口愈合试验和Transwell试验评估迁移能力。为进一步研究miR-221-5p在人前列腺癌中的作用,我们建立了体内裸鼠异种移植模型。
miR-221-5p通过靶向SOCS1的3'UTR调控其表达,从而在体外调节前列腺癌细胞的增殖、迁移以及在体内调节肿瘤发生,并且miR-221-5p调节前列腺癌细胞中的MAPK/ERK信号通路和EMT特征。
前列腺癌细胞中miR-221-5p表达的上调和沉默与细胞增殖、迁移和肿瘤发生相关,这表明miR-221-5p在前列腺癌进展中起重要作用。