Department of Pathology, Tangshan Gongren Hospital, Tangshan, Hebei, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5391-5403. doi: 10.26355/eurrev_202005_21323.
Esophageal squamous cell carcinoma (ESCC) is a common malignant epithelial tumor in the elderly, and the cause is very complicated. Therefore, the study of the pathogenesis of ESCC is conducive to the effective treatment of ESCC. Many studies indicated that lncRNAs were important regulatory factors in tumor formation and disease development. However, the regulatory network of lncRNA in ESCC has not been fully explored.
The expression of miR-574-3p, ZEB2-AS1, and HMGA2 was measured using qRT-PCR. The protein expression of PCNA, Cleaved-caspase3, MMP9, and HMGA2 was detected through Western blot. Cell proliferation or apoptosis of transfected cells was calculated via CKK-8 assay or flow cytometry. Transwell assay was applied to detect cell migration and invasion of ESCC cells. Luciferase reporter assay and RNA pull-down were used to determine the relationship among miR-574-3p, ZEB2-AS1, and HMGA2 in ESCC. Moreover, the regulatory network of ZEB2-AS1 has been verified in vivo in this study.
We found that ZEB2-AS1 was upregulated in ESCC tissues and cells. The knockdown of ZEB2-AS1 could inhibit cell proliferation, invasion, and migration, as well as promoted cell apoptosis in ESCC. Interestingly, miR-574-3p deficiency or HMGA2 promotion could reverse the effects of si-ZEB2-AS1 on ESCC cell progression. Luciferase reporter assay indicated that miR-574-3p was a target miRNA of ZEB2-AS1 and HMGA2 was a target gene of miR-574-3p in ESCC.
In this paper, we first verified the novel regulatory mechanism of lncRNA ZEB2-AS1 in ESCC cellular process. LncRNA ZEB2-AS1 promoted the proliferation, migration, and invasion of ESCC by modulating miR-574-3p/HMGA2 axis, indicating that ZEB2-AS1 played essential roles in cell progression in ESCC and providing a new therapeutic target of ESCC.
食管鳞状细胞癌（ESCC）是老年人常见的恶性上皮肿瘤，其病因非常复杂。因此，研究 ESCC 的发病机制有助于 ESCC 的有效治疗。许多研究表明，lncRNAs 是肿瘤形成和疾病发展的重要调节因子。然而，lncRNA 在 ESCC 中的调控网络尚未得到充分探索。
采用 qRT-PCR 检测 miR-574-3p、ZEB2-AS1 和 HMGA2 的表达。通过 Western blot 检测 PCNA、Cleaved-caspase3、MMP9 和 HMGA2 的蛋白表达。通过 CCK-8 测定或流式细胞术计算转染细胞的增殖或凋亡。采用 Transwell 测定法检测 ESCC 细胞的迁移和侵袭。利用荧光素酶报告基因和 RNA 下拉实验来确定 miR-574-3p、ZEB2-AS1 和 HMGA2 之间在 ESCC 中的关系。此外，本研究还在体内验证了 ZEB2-AS1 的调控网络。
我们发现 ZEB2-AS1 在 ESCC 组织和细胞中上调。ZEB2-AS1 的敲低可抑制 ESCC 细胞的增殖、侵袭和迁移，并促进细胞凋亡。有趣的是，miR-574-3p 的缺失或 HMGA2 的促进作用可逆转 si-ZEB2-AS1 对 ESCC 细胞进展的影响。荧光素酶报告基因实验表明，miR-574-3p 是 ZEB2-AS1 的靶 miRNA，HMGA2 是 ESCC 中的靶基因。
本文首次验证了 lncRNA ZEB2-AS1 在 ESCC 细胞过程中的新的调控机制。LncRNA ZEB2-AS1 通过调节 miR-574-3p/HMGA2 轴促进 ESCC 的增殖、迁移和侵袭，表明 ZEB2-AS1 在 ESCC 细胞进展中发挥重要作用，并为 ESCC 提供了新的治疗靶点。
