Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China.
Department of General Surgery, Bazhong Central Hospital, Bazhong 636000, China.
Acta Biochim Biophys Sin (Shanghai). 2021 May 21;53(6):652-662. doi: 10.1093/abbs/gmab040.
Cancer stem cells (CSCs) are major contributors to tumor initiation, recurrence, and metastasis of hepatocellular carcinoma (HCC). Some long non-coding RNAs have been reported as modulators of stem-like properties in cancer cells. However, the role of LINC01013 in liver CSCs has not yet been clarified. In this study, we aimed to elucidate the expression pattern and functions of LINC01013 in HCC. HCC tissues and normal controls were collected, and the expression pattern of LINC01013 and miR-6795-5p was identified by quick real-time polymerase chain reaction. Cell counting kit-8 assay, colony formation, and spheroid formation were performed to measure cell viability, proliferation, and self-renewal of HCC cell lines. The expression of stem markers was detected by western blot analysis. The effect of LINC01013 on viability, proliferation, and stem-like properties was detected through gain-of-function and loss-of-function experiments. The direct interaction among LINC01013, miR-6795-5p, and FMNL3 was testified by dual-luciferase reporter gene assay. Tumor-bearing mice were constructed to ascertain the functions of LINC01013 in vivo. HCC tissues showed increased LINC01013 and FMNL3 expression, while it showed a decreased miR-6795-5p expression as compared to the relative controls. Moreover, the high level of LINC01013 was closely related to the poor prognosis of HCC patients. LINC01013 directly binds to miR-6795-5p and subsequently relieves FMNL3. Silencing LINC01013, FMNL3, or overexpression of miR-6795-5p could suppress spheroid and colony formation, proliferation, as well as expression of stemness markers in HepG2 and SNU-182 cells. LINC01013 knockdown suppressed growth and stem-like traits of HCC cells in vivo by reducing FMNL3 expression. LINC01013/miR-6795-5p/FMNL3 axis may be a novel therapeutic target for HCC.
癌症干细胞 (CSCs) 是肿瘤起始、复发和转移的主要贡献者。一些长非编码 RNA 已被报道为癌细胞干性特征的调节剂。然而，LINC01013 在肝 CSCs 中的作用尚未阐明。在这项研究中，我们旨在阐明 LINC01013 在 HCC 中的表达模式和功能。收集 HCC 组织和正常对照，通过快速实时聚合酶链反应鉴定 LINC01013 和 miR-6795-5p 的表达模式。通过细胞计数试剂盒-8 测定、集落形成和球体形成来测量 HCC 细胞系的细胞活力、增殖和自我更新。通过 Western blot 分析检测干细胞标志物的表达。通过功能获得和功能丧失实验检测 LINC01013 对活力、增殖和干性特征的影响。通过双荧光素酶报告基因测定验证 LINC01013、miR-6795-5p 和 FMNL3 之间的直接相互作用。构建荷瘤小鼠以确定 LINC01013 在体内的功能。与相对对照相比，HCC 组织中 LINC01013 和 FMNL3 的表达增加，而 miR-6795-5p 的表达降低。此外，LINC01013 的高水平与 HCC 患者的不良预后密切相关。LINC01013 直接与 miR-6795-5p 结合，随后缓解 FMNL3。沉默 LINC01013、FMNL3 或过表达 miR-6795-5p 可抑制 HepG2 和 SNU-182 细胞的球体和集落形成、增殖以及干性标志物的表达。LINC01013 敲低通过降低 FMNL3 表达抑制 HCC 细胞在体内的生长和干性特征。LINC01013/miR-6795-5p/FMNL3 轴可能是 HCC 的一种新的治疗靶点。
