Digestive System Department, Yankuang new journey general hospital, Jining City, China.
Qingdao central hospital, Qingedao City, China.
J Gene Med. 2020 Sep;22(9):e3202. doi: 10.1002/jgm.3202. Epub 2020 May 28.
Long non-coding RNAs (lncRNAs) function as oncogenes or tumor suppressor genes in several cancers. The present study aimed to determine the functions of lncRNA HOXC-AS1 in gastric cancer (GC) in vitro.
A quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to measure the expression of lncRNA HOXC-AS1 in GC cell lines and normal cells. After silencing HOXC-AS1 in GC cells, a cell counting kit-8 assay monitored the viability of the cells. qRT-PCR and western blot documented the EMT key genes in response to HOXC-AS1 change. qRT-PCR detected mRNA expression for eIF4AIII in GC and normal cell lines and cell viability was measured after an increase and decrease of eIF4AIII. RNA pull-down and qRT-PCR confirmed the binding in between. Apoptosis was compared by flow cytometry. The interplay between the two genes was surveyed by introduction of the sh-HOXC-AS1 and sh-eIF4AIII and by assessing cell viability, EMT and Wnt/β-catenin signaling.
lncRNA HOXC-AS1 expression is up-regulated in GC cells and a decrease of lncRNA HOXC-AS1 inhibited cell viability. Binding was validated by RNA pull-down. Additionally, inhibition of eIF4AIII induced an increase of lncRNA HOXC-AS1, thus promoting cell proliferation and the EMT process but deterring apoptosis of gastric cancer cells. Wnt/β-catenin signaling was impeded by HOXC-AS1 inhibition but restored by suppression of eIF4AIII.
HOXC-AS1 may promote the proliferation and the EMT process and inhibit apoptosis by binding eIF4AIII via Wnt/β-catenin signaling, which indicates that HOXC-AS1/eIF4AIII might be an axis that could be further used as a biomarker to help with the diagnosis of GC.
长链非编码 RNA(lncRNA)在几种癌症中作为癌基因或肿瘤抑制基因发挥作用。本研究旨在确定 lncRNA HOXC-AS1 在体外胃癌(GC)中的功能。
采用实时定量逆转录聚合酶链反应(qRT-PCR)检测 lncRNA HOXC-AS1 在 GC 细胞系和正常细胞中的表达。沉默 GC 细胞中的 HOXC-AS1 后,细胞计数试剂盒-8 检测细胞活力。qRT-PCR 和 Western blot 记录 EMT 关键基因对 HOXC-AS1 变化的反应。qRT-PCR 检测 GC 和正常细胞系中 eIF4AIII 的 mRNA 表达,然后增加和减少 eIF4AIII 后测量细胞活力。RNA 下拉和 qRT-PCR 证实了两者之间的结合。通过流式细胞术比较细胞凋亡。通过引入 sh-HOXC-AS1 和 sh-eIF4AIII 并评估细胞活力、EMT 和 Wnt/β-catenin 信号转导来研究这两个基因之间的相互作用。
lncRNA HOXC-AS1 在 GC 细胞中表达上调,lncRNA HOXC-AS1 减少抑制细胞活力。RNA 下拉验证了结合。此外,抑制 eIF4AIII 会增加 lncRNA HOXC-AS1,从而促进胃癌细胞的增殖和 EMT 过程,但抑制细胞凋亡。Wnt/β-catenin 信号转导被 HOXC-AS1 抑制抑制,但被 eIF4AIII 抑制恢复。
HOXC-AS1 可能通过 Wnt/β-catenin 信号转导与 eIF4AIII 结合促进增殖和 EMT 过程并抑制凋亡,这表明 HOXC-AS1/eIF4AIII 可能是一个轴,可进一步用作帮助诊断 GC 的生物标志物。