Silencing of long noncoding RNA HOXA11-AS inhibits the Wnt signaling pathway via the upregulation of HOXA11 and thereby inhibits the proliferation, invasion, and self-renewal of hepatocellular carcinoma stem cells
Silencing of long noncoding RNA HOXA11-AS inhibits the Wnt signaling pathway via the upregulation of HOXA11 and thereby inhibits the proliferation, invasion, and self-renewal of hepatocellular carcinoma stem cells
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DOI：
10.1038/s12276-019-0328-x
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发表时间：
2019-11-22
影响因子：
12.8
通讯作者：
Zhuo, Liu
中科院分区：
文献类型：
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作者：
Guo, Jun-Cheng;Yang, Yi-Jun;Zhuo, Liu
肝细胞癌（HCC）是癌症相关死亡的主要原因，但其分子机制尚未得到很好的表征。长链非编码RNA（lncRNA）在肿瘤发生中起着重要作用，包括肝癌的发生。然而，同源框A11反义（HOXA 11-AS）在决定肝癌干细胞特性中的作用仍有待解释，因此，本研究旨在研究HOXA 11-AS对肝癌干细胞特性的影响。首先，确定了HCC组织、细胞和干细胞中HOXA 11-AS和HOXA 11的表达模式。从Hep 3B和Huh 7细胞中成功分选出肝癌干细胞，分别转染短发夹状或过表达质粒HOXA 11-AS或HOXA 11过表达和缺失，研究这些介质对肝癌干细胞自我更新、增殖、迁移和体内致瘤性的影响。此外，通过Dickkopf-1（Wnt信号通路抑制剂）治疗，探索了HOXA 11-AS、HOXA 11和Wnt信号通路之间的潜在关系和调控机制。肝癌干细胞高表达HOXA 11-AS，低表达HOXA 11。HOXA 11-AS沉默和HOXA 11过表达均抑制体内HCC干细胞的自我更新、增殖、迁移和致瘤性，如癌症干细胞表面标志物（CD 133和CD 44）和干细胞相关转录因子（Nanog、Sox 2和Oct 4）表达降低所证明。此外，沉默HOXA 11-AS通过降低HOXA 11启动子的甲基化水平而使Wnt信号通路失活，从而抑制HCC干细胞特征。总的来说，这项研究表明，HOXA 11-AS沉默发挥抗肿瘤作用，通过降低HOXA 11启动子的甲基化水平，通过Wnt信号通路失活抑制HCC的发展。
Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths, but its molecular mechanisms are not yet well characterized. Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis, including that of HCC. However, the role of homeobox A11 antisense (HOXA11-AS) in determining HCC stem cell characteristics remains to be explained; hence, this study aimed to investigate the effects of HOXA11-AS on HCC stem cell characteristics. Initially, the expression patterns of HOXA11-AS and HOXA11 in HCC tissues, cells, and stem cells were determined. HCC stem cells, successfully sorted from Hep3B and Huh7 cells, were transfected with short hairpin or overexpression plasmids for HOXA11-AS or HOXA11 overexpression and depletion, with an aim to study the influences of these mediators on the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo. Additionally, the potential relationship and the regulatory mechanisms that link HOXA11-AS, HOXA11, and the Wnt signaling pathway were explored through treatment with Dickkopf-1 (a Wnt signaling pathway inhibitor). HCC stem cells showed high expression of HOXA11-AS and low expression of HOXA11. Both HOXA11-AS silencing and HOXA11 overexpression suppressed the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo, as evidenced by the decreased expression of cancer stem cell surface markers (CD133 and CD44) and stemness-related transcription factors (Nanog, Sox2, and Oct4). Moreover, silencing HOXA11-AS inactivated the Wnt signaling pathway by decreasing the methylation level of the HOXA11 promoter, thereby inhibiting HCC stem cell characteristics. Collectively, this study suggested that HOXA11-AS silencing exerts an antitumor effect, suppressing HCC development via Wnt signaling pathway inactivation by decreasing the methylation level of the HOXA11 promoter.