Novel role of LINC01013/miR-6795-5p/FMNL3 axis in the regulation of hepatocellular carcinoma stem cell features
Novel role of LINC01013/miR-6795-5p/FMNL3 axis in the regulation of hepatocellular carcinoma stem cell features
复制标题
LINC01013/miR-6795-5p/FMNL3轴在调控肝细胞癌干细胞特征中的新作用
DOI：
10.1093/abbs/gmab040
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发表时间：
2021
影响因子：
3.7
通讯作者：
Wang Bo
中科院分区：
文献类型：
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作者：
Wang Wanli;Xu Shicheng;Di Ying;Zhang Zhiyong;Li Qingshan;Guo Kun;Lv Yi;Wang Bo
肿瘤干细胞（cancer stem cells，CSCs）是肝细胞癌（hepatocellular carcinoma，HCC）发生、复发和转移的主要因素。一些长的非编码RNA已被报道为癌细胞中干细胞样性质的调节剂。然而，LINC 01013在肝CSC中的作用尚未阐明。本研究旨在阐明LINC 01013在肝癌中的表达模式和功能。收集肝癌组织和正常对照，采用快速实时荧光定量PCR方法检测LINC 01013和miR-6795- 5 p的表达情况。进行细胞计数试剂盒-8测定、集落形成和球体形成以测量HCC细胞系的细胞活力、增殖和自我更新。Western blot分析茎标记的表达。通过功能获得和功能丧失实验检测LINC 01013对活力、增殖和干细胞样性质的影响。通过双荧光素酶报告基因分析证实LINC 01013、miR-6795- 5 p和FMNL 3之间的直接相互作用。构建荷瘤小鼠以确定LINC 01013在体内的功能。与相对对照相比，HCC组织显示出增加的LINC 01013和FMNL 3表达，而其显示出降低的miR-6795- 5 p表达。此外，LINC 01013的高表达与HCC患者的不良预后密切相关。LINC 01013直接与miR-6795- 5 p结合，随后缓解FMNL 3。沉默LINC 01013、FMNL 3或过表达miR-6795- 5 p可以抑制HepG 2和SNU-182细胞中球状体和集落的形成、增殖以及干性标志物的表达。LINC 01013敲低通过降低FMNL 3表达抑制体内HCC细胞的生长和干细胞样特征。LINC 01013/miR-6795- 5 p/FMNL 3轴可能成为肝癌治疗的新靶点。
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.