肿瘤干细胞(CSC)是肿瘤发生过程中的关键调节因子。programming,和复发。保持其干性的机制仍然是神秘的,尽管一些长链非编码RNA(lncRNA)的作用已在胰腺癌干细胞(PCSCs)中得到强调。在这项研究中,我们首先确定PCSCs过表达lncRNANORAD,然后研究了NORAD对PCSC干性维持的影响。
lncRNANORAD的表达,在RNA分离后定量PC组织和细胞系中的miR-202-5p和ANP32E。双荧光素酶报告基因测定,进行RNA下拉和RIP测定以验证NORAD之间的相互作用,miR-202-5p和ANP32E。然后我们进行了miR-202-5p的功能增益和丧失,ANP32E和NORAD在PANC-1细胞系中,然后测量醛脱氢酶活性,细胞活力,凋亡,细胞周期分布,菌落形成,PC细胞的自我更新能力和致瘤性。
LncRNANORAD和ANP32E在PC组织和细胞中上调,而miR-202-5p水平下调。LncRNANORAD竞争性结合miR-202-5p,并促进miR-202-5p靶基因ANP32E的表达,从而促进PC细胞的活力,扩散,和体外自我更新能力,以及促进PCSCs在体内的肿瘤发生。
总的来说,lncRNANORAD通过竞争性结合miR-202-5上调ANP32E表达,从而加速PCSCs的增殖和自我更新。
Cancer stem cells (CSCs) are key regulators in the processes of tumor initiation, progression, and recurrence. The mechanism that maintains their stemness remains enigmatic, although the role of several long noncoding RNAs (lncRNAs) has been highlighted in the pancreatic cancer stem cells (PCSCs). In this study, we first established that PCSCs overexpressing lncRNA NORAD, and then investigated the effects of NORAD on the maintenance of PCSC stemness.
Expression of lncRNA NORAD, miR-202-5p and
ANP32E in PC tissues and cell lines was quantified after RNA isolation. Dual-luciferase reporter assay, RNA pull-down and RIP assays were performed to verify the interactions among NORAD, miR-202-5p and
ANP32E. We then carried out gain- and loss-of function of miR-202-5p, ANP32E and NORAD in PANC-1 cell line, followed by measurement of the aldehyde dehydrogenase activity, cell viability, apoptosis, cell cycle distribution, colony formation, self-renewal ability and tumorigenicity of PC cells.
LncRNA NORAD and
ANP32E were upregulated in PC tissues and cells, whereas the miR-202-5p level was down-regulated. LncRNA NORAD competitively bound to miR-202-5p, and promoted the expression of the miR-202-5p target gene
ANP32E thereby promoting PC cell viability, proliferation, and self-renewal ability in vitro, as well as facilitating tumorigenesis of PCSCs in vivo.
Overall, lncRNA NORAD upregulates
ANP32E expression by competitively binding to miR-202-5, which accelerates the proliferation and self-renewal of PCSCs.