ABSTRACT

We intended to investigate the underlying mechanism of action of long noncoding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) in colorectal cancer (CRC) progression, especially in tumor cell stemness. For that purpose, different assays were performed such as real-time PCR and western blotting to determine the expression of target genes. Cell stemness was determined by sphere formation assay, flow cytometry assay, and the analysis of stemness‐related markers. The interplay among target genes was evaluated using bioinformatics analyses, luciferase reporter and biotin-labeled RNA pull down assays. We found that HOTAIR was highly expressed and predicted poor prognosis survival in CRC. Downregulation of HOTAIR repressed tumor malignant behaviors and cancer stemness. Mechanistically, HOTAIR facilitated the expression of the microRNA (miR)-211-5p target gene fms-like tyrosine kinase-1 (FLT-1), thereby modulating cancer stem cell (CSC) properties in CRC. We conclude that HOTAIR/miR-211-5p/FLT-1 axis contributes to CRC cancer stemness.

摘要

我们打算研究长链非编码 RNA (lncRNA) HOX 转录反义 RNA (HOTAIR) 在结直肠癌 (CRC) 进展中的潜在作用机制，尤其是在肿瘤细胞干性中。为此目的，进行了不同的测定，例如实时 PCR 和蛋白质印迹以确定靶基因的表达。通过球体形成测定、流式细胞术测定和干性相关标志物分析确定细胞干性。使用生物信息学分析、荧光素酶报告基因和生物素标记的 RNA 下拉分析评估靶基因之间的相互作用。我们发现 HOTAIR 在 CRC 中高度表达并预测预后不良。HOTAIR的下调抑制了肿瘤恶性行为和癌症干性。机械地，HOTAIR 促进了 microRNA (miR)-211-5p 靶基因 fms 样酪氨酸激酶-1 (FLT-1) 的表达，从而调节 CRC 中的癌症干细胞 (CSC) 特性。我们得出结论，HOTAIR/miR-211-5p/FLT-1 轴有助于 CRC 癌症干性。