Niu Yulin, He Jin Hua, Zhang Yinglian, Li Kun, Xing Chungen
Department of Transplantation Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China.
Department of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, Guangdong 511400, P.R. China.
Oncol Lett. 2021 Jun;21(6):469. doi: 10.3892/ol.2021.12730. Epub 2021 Apr 13.
Prostate cancer is one of the most common malignant tumors in men. Patients with local infiltration and distant metastasis often have a poor prognosis. The present study aimed to investigate the expression and regulatory mechanism of the circular RNA cerebellar degeneration-related protein 1, anti-sense (circCDR1as) in prostate cancer cell lines. MicroRNAs (miRNAs) regulated by circCDR1as and target genes regulated by miRNAs were predicted using bioinformatics software. Prostate cancer cell lines (LNCaP, 22Rv1 and PC-3), a normal prostate epithelial cell line (RWPE-1) and a human embryonic kidney cell line (293T) were cultured. Relative gene expression was detected using reverse transcription PCR. Small interfering RNAs (siRNAs) targeting circCDR1as and X-linked inhibitor of apoptosis protein (XIAP) and miRNA mimics were designed and transfected into the cell lines using Lipofectamine 3000. Cell invasion was determined using a Transwell assay, the cell proliferation rate was detected using an MTT assay and cell migration was examined using a scratch assay. Relative protein expression was detected using western blotting. Double fluorescent reporter gene vectors and an anti-Ago2 RNA-binding protein immunoprecipitation assay were used to verify binding. Bioinformatics analyses indicated that there was a binding site between miR-641 and circCDR1as and between miR-641 and XIAP. The expression of circCDR1as and XIAP was higher and the expression of miR-641 was lower in the prostate cancer cell lines compared with the normal prostate epithelial cell line. After effectively reducing the expression of circCDR1as and XIAP and increasing the expression of miR-641 in PC-3 cells, the proliferation, invasion and migration of PC-3 cells were effectively inhibited. circCDR1as could bind to miR-641, which targeted the 3'-untranslated region of XIAP. Reducing the expression of circCDR1 promoted the expression of miR-641 and inhibited the expression of XIAP. Overall, the circCDR1as/miR-641/XIAP regulatory axis plays a role in the invasion and migration of the prostate cancer PC-3 cell line.
前列腺癌是男性最常见的恶性肿瘤之一。出现局部浸润和远处转移的患者预后往往较差。本研究旨在探究环状RNA小脑变性相关蛋白1反义转录本(circCDR1as)在前列腺癌细胞系中的表达及调控机制。利用生物信息学软件预测受circCDR1as调控的微小RNA(miRNA)以及受miRNA调控的靶基因。培养前列腺癌细胞系(LNCaP、22Rv1和PC-3)、正常前列腺上皮细胞系(RWPE-1)和人胚肾细胞系(293T)。采用逆转录PCR检测相关基因表达。设计靶向circCDR1as和X连锁凋亡抑制蛋白(XIAP)的小干扰RNA(siRNA)以及miRNA模拟物,并用Lipofectamine 3000转染细胞系。采用Transwell实验测定细胞侵袭能力,采用MTT实验检测细胞增殖率,采用划痕实验检测细胞迁移能力。采用蛋白质免疫印迹法检测相关蛋白表达。利用双荧光报告基因载体和抗Ago2 RNA结合蛋白免疫沉淀实验验证结合情况。生物信息学分析表明,miR-641与circCDR1as之间以及miR-641与XIAP之间存在结合位点。与正常前列腺上皮细胞系相比,前列腺癌细胞系中circCDR1as和XIAP的表达较高,而miR-641的表达较低。在PC-3细胞中有效降低circCDR1as和XIAP的表达并提高miR-641的表达后,PC-3细胞的增殖、侵袭和迁移能力受到有效抑制。circCDR1as可与miR-641结合,而miR-641靶向XIAP的3'-非翻译区。降低circCDR1的表达可促进miR-641的表达并抑制XIAP的表达。总体而言,circCDR1as/miR-641/XIAP调控轴在前列腺癌PC-3细胞系的侵袭和迁移中发挥作用。