Zheng Xinzi, Cui Renyin, Jiao Yan, Chu Dongxia, Wang Bingrong, Li Na
Department of Pathophysiology, College of Basic Medical Sciences, Jilin University, Changchun, Jilin, P.R. China.
Department of Hepatobiliary and Pancreatic Surgery, The First Hospital of Jilin University, Changchun, Jilin, P.R. China.
Folia Histochem Cytobiol. 2024;62(4):165-179. doi: 10.5603/fhc.100686. Epub 2024 Nov 26.
Increasing evidence shows that Hox transcript antisense RNA (HOTAIR) plays a vital role in liver cancer initiation and progression by affecting the proliferation, invasion, migration, and apoptosis of liver cancer cells. However, the underlying mechanism of how HOTAIR exerts its functions in liver cancer cells remains unclear. Previous studies have shown that HOTAIR affects the invasion and migration of liver cancer cells by regulating the expression of E-cadherin. Snail2, a transcription factor involved in epithelial-mesenchymal transition, directly binds to the E-boxes of the E-cadherin promoter to repress its transcription. The aim of the study was to examine the correlation between HOTAIR and Snail2 in the HOTAIR/Snail2/E-cadherin signal pathway and explore the role of HOTAIR in the proliferation, invasion, migration, and apoptosis of liver cancer cells.
Fifty matched normal liver tissues and 373 liver cancer tissues were analysed and evaluated. HepG2 and SNU-387 cells were cultured and transfected with plasmids knocking down HOTAIR to disrupt HOTAIR expression. Cell scratch and transwell assays were performed to examine the migration and invasion of HepG2 and SNU-387 cells; in addition, the expression of MMP2 and MMP9 was detected by immunoblotting analysis, RT-qPCR analysis, immunofluorescence analysis, and bioinformatics analysis, which elucidated the regulatory relationship between HOTAIR and Snail2. We used flow cytometry and JC-1 probe analysis assays to clarify the function of HOTAIR inliver cancer cell apoptosis.
The HOTAIR mRNA was upregulated in liver cancer tissues, which was related to worse overall survival. HOTAIR induced the expression of matrix metalloproteinase-9 (MMP9) and metalloproteinase-2 (MMP2), leading to degradation of extracellular matrix. HOTAIR knockdown significantly reduced the doubling time and inhibited cell migration and invasion of liver cancer cells. Furthermore, HOTAIR depletion induced mitochondrial-related apoptosis in HepG2 and SNU-387 cell lines.
In this study, we propose a novel mechanism in which HOTAIR promotes invasion and migration of liver cancer cells by regulating the nuclear localisation of Snail2.
越来越多的证据表明，Hox转录本反义RNA（HOTAIR）通过影响肝癌细胞的增殖、侵袭、迁移和凋亡，在肝癌的发生和发展中起着至关重要的作用。然而，HOTAIR在肝癌细胞中发挥功能的潜在机制仍不清楚。先前的研究表明，HOTAIR通过调节E-钙黏蛋白的表达来影响肝癌细胞的侵袭和迁移。Snail2是一种参与上皮-间质转化的转录因子，它直接与E-钙黏蛋白启动子的E盒结合，从而抑制其转录。本研究的目的是检测HOTAIR/Snail2/E-钙黏蛋白信号通路中HOTAIR与Snail2之间的相关性，并探讨HOTAIR在肝癌细胞增殖、侵袭、迁移和凋亡中的作用。
对50对匹配的正常肝组织和373例肝癌组织进行分析和评估。培养HepG2和SNU-387细胞，并用敲低HOTAIR的质粒进行转染，以破坏HOTAIR的表达。进行细胞划痕和Transwell实验，检测HepG2和SNU-387细胞的迁移和侵袭能力；此外，通过免疫印迹分析、RT-qPCR分析、免疫荧光分析和生物信息学分析检测MMP2和MMP9的表达，以阐明HOTAIR与Snail2之间的调控关系。我们使用流式细胞术和JC-1探针分析实验来阐明HOTAIR在肝癌细胞凋亡中的作用。
肝癌组织中HOTAIR mRNA表达上调，这与较差的总生存期相关。HOTAIR诱导基质金属蛋白酶-9（MMP9）和金属蛋白酶-2（MMP2）的表达，导致细胞外基质降解。敲低HOTAIR显著延长了肝癌细胞的倍增时间，并抑制了其迁移和侵袭能力。此外，敲低HOTAIR可诱导HepG2和SNU-387细胞系发生线粒体相关凋亡。
在本研究中，我们提出了一种新的机制，即HOTAIR通过调节Snail2的核定位来促进肝癌细胞的侵袭和迁移。
