Chen Ping-Ping, Zhang Zhen-Sheng, Wu Jin-Cai, Zheng Jin-Fang, Lin Fan
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Jinan University, Guangzhou 510630, Guangdong Province, PR China; Department of Hepatobiliary Surgery, Hainan Provincial People's Hospital, Haikou 570311, Hainan Province, PR China.
Department of Hepatobiliary Surgery, Hainan Provincial People's Hospital, Haikou 570311, Hainan Province, PR China.
Cell Signal. 2021 Aug;84:109992. doi: 10.1016/j.cellsig.2021.109992. Epub 2021 Mar 24.
Hepatocellular carcinoma (HCC) is the most common cancer and its prognosis is poor due to metastasis and recurrence. EMT is associated with metastasis. A deep understanding of regulatory mechanism of EMT is critical. LncRNA is involved in regulation of various biological processes including EMT. This study aimed to investigate the regulatory signal axis among lncRNA SNHG12, miR-516a-5p and the target gene HEG1 during EMT. Cell cycle and apoptosis were analyzed by flow cytometry. Tumorigenesis was analyzed by clone formation assay. Wound healing assay and transwell assay was performed to detect migration and invasion, respectively. Interaction among SNHG12, miR-516a-5p and HEG1 were analyzed by dual luciferase assay and RIP assay. We also detected expression of RNA and protein by QPCR and western blotting. Finally, tumor growth was analyzed by tumorigenesis assay in vivo. Ki-67 and HEG1 level in tumor tissues was analyzed by IHC. SNHG12 and HEG1 were upregulated, miR-516a-5p was downregulated in HCC cell lines. SNHG12 could interact with and inhibit miR-516a-5p. MiR-516a-5p could interact with HEG1 and inhibit HEG1 expression. Knock down SNHG12 inhibited proliferation, migration, invasion, EMT and promoted apoptosis of HCC cells. Such effects were antagonized by inhibiting miR-516a-5p. SNHG12 overexpression lead to opposite results. Similar results were observed in mice. SNHG12 could promote EMT in HCC through targeting and inhibiting miR-516a-5p, which eventually upregulated HEG1 expression, in both cell and mice.
肝细胞癌（HCC）是最常见的癌症，由于转移和复发，其预后较差。上皮-间质转化（EMT）与转移有关。深入了解EMT的调控机制至关重要。长链非编码RNA（lncRNA）参与包括EMT在内的各种生物学过程的调控。本研究旨在探讨lncRNA SNHG12、miR-516a-5p与靶基因HEG1在EMT过程中的调控信号轴。通过流式细胞术分析细胞周期和凋亡。通过克隆形成试验分析肿瘤发生。分别进行伤口愈合试验和Transwell试验以检测迁移和侵袭。通过双荧光素酶试验和RNA免疫沉淀试验（RIP试验）分析SNHG12、miR-516a-5p和HEG1之间的相互作用。我们还通过定量聚合酶链反应（QPCR）和蛋白质印迹法检测RNA和蛋白质的表达。最后，通过体内肿瘤发生试验分析肿瘤生长。通过免疫组织化学（IHC）分析肿瘤组织中的Ki-67和HEG1水平。在肝癌细胞系中，SNHG12和HEG1上调，miR-516a-5p下调。SNHG12可与miR-516a-5p相互作用并抑制其表达。miR-516a-5p可与HEG1相互作用并抑制HEG1表达。敲低SNHG12可抑制肝癌细胞的增殖、迁移、侵袭、EMT并促进其凋亡。抑制miR-516a-5p可拮抗这些作用。SNHG12过表达导致相反结果。在小鼠中观察到类似结果。在细胞和小鼠中，SNHG12均可通过靶向并抑制miR-516a-5p促进肝癌中的EMT，最终上调HEG1表达。
