Wang Ke, Nie Yuanhua, Wang Shilong, Chang Dongmin
Department of Surgical Oncology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.
J Biochem Mol Toxicol. 2025 Jan;39(1):e70104. doi: 10.1002/jbt.70104.
Increasing long noncoding RNAs (lncRNAs) have been found to participate in regulating the progression of colorectal cancer (CRC), which is a common gastrointestinal malignancy. Here, the specific role and mechanisms of lncRNA LINC00294 were investigated in CRC. The expression levels of LINC00294, miR-499a-5p, and La-related protein 4B (LARP4B) in CRC cells (HCT116 and SW620) and tissues were assessed by RT-qPCR. The viability, proliferation, cell cycle, and apoptosis of HCT116 and SW620 cells were determined by CCK-8, EdU, and flow cytometry assays. Protein levels of cell cycle and cell apoptosis markers were measured by western blot analysis. FISH assay was performed to evaluate the subcellular localization of LINC00294 in CRC cells. Luciferase reporter assay, RNA pull-down assay, as well as RIP assay verified the interactions between miR-499a-5p and LINC00294 (or LARP4B). The xenograft model was established in mice to investigate the function of LINC00294 in vivo. LINC00294 presented a decreased expression level in CRC cells and tissues. LINC00294 overexpression suppressed the cell proliferative capacity, promoted cell cycle arrest, and induced cell apoptosis in CRC HCT116 and SW620 cells. LINC00294 interacted with miR-499a-5p, and miR-499a-5p targeted LARP4B. MiR-499a-5p was upregulated while LARP4B was downregulated in CRC cells. In rescue assays, LARP4B knockdown reversed the inhibition of LINC00294 overexpression on malignant phenotypes of HCT116 and SW620 cells. In the xenograft model, LINC00294 overexpression inhibited tumor growth in vivo. LINC00294 exerted an antitumor function in CRC by forming a LINC00294/miR-499a-5p/LARP4B regulatory network.
越来越多的长链非编码RNA（lncRNA）被发现参与调控结直肠癌（CRC）的进展，结直肠癌是一种常见的胃肠道恶性肿瘤。在此，研究了lncRNA LINC00294在结直肠癌中的具体作用及机制。通过RT-qPCR评估结直肠癌细胞（HCT116和SW620）及组织中LINC00294、miR-499a-5p和La相关蛋白4B（LARP4B）的表达水平。通过CCK-8、EdU和流式细胞术检测法测定HCT116和SW620细胞的活力、增殖、细胞周期及凋亡情况。通过蛋白质印迹分析检测细胞周期和细胞凋亡标志物的蛋白质水平。进行荧光原位杂交检测法以评估LINC00294在结直肠癌细胞中的亚细胞定位。荧光素酶报告基因检测法、RNA下拉检测法以及RNA免疫沉淀检测法验证了miR-499a-5p与LINC00294（或LARP4B）之间的相互作用。在小鼠体内建立异种移植模型以研究LINC00294在体内的功能。LINC00294在结直肠癌细胞及组织中的表达水平降低。LINC00294过表达抑制了结直肠癌HCT116和SW620细胞的增殖能力，促进细胞周期停滞并诱导细胞凋亡。LINC00294与miR-499a-5p相互作用，且miR-499a-5p靶向LARP4B。在结直肠癌细胞中，miR-499a-5p上调而LARP4B下调。在拯救实验中，敲低LARP4B可逆转LINC00294过表达对HCT116和SW620细胞恶性表型的抑制作用。在异种移植模型中，LINC00294过表达在体内抑制肿瘤生长。LINC00294通过形成LINC00294/miR-499a-5p/LARP4B调控网络在结直肠癌中发挥抗肿瘤作用。
