Department of Cardiology, First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Eur Rev Med Pharmacol Sci. 2018 Oct;22(20):6983-6990. doi: 10.26355/eurrev_201810_16169.
To investigate whether HOX transcript antisense RNA (HOTAIR) regulates myocardial fibrosis via promoting proliferation of cardiac fibroblasts (CFs) and upregulating the expression levels of fibrotic proteins through activating Wnt signaling pathway.
The expression level of HOTAIR in Ang II-induced cardiac fibroblasts was detected by quantitative Real-time-polymerase chain reaction (qRT-PCR). Overexpression or knockdown of HOTAIR expression was achieved by lentivirus transfection. The effects of HOTAIR on regulating cell proliferation, migration and apoptosis were measured by cell counting kit-8 (CCK-8) test, transwell assay and flow cytometry, respectively. Western blot and qRT-PCR experiments were performed to detect expressions of fibrosis-related genes and Wnt pathway-related genes. Target gene of HOTAIR was predicted by bioinformatics analysis. Rescue assays were conducted to assess whether HOTAIR could regulate cell proliferation and fibrosis by activating Wnt signaling pathway via URI1.
QRT-PCR results showed that HOTAIR expression in Ang II-induced CF cells was significantly higher than that in control. HOTAIR overexpression in CF cells can promote cell proliferation and migration, inhibit apoptosis, and promote the expressions of fibrosis-related genes. Western blot results indicated that HOTAIR could upregulate URI1 expression and activate Wnt signaling pathway. In addition, rescue assay demonstrated that overexpression of URI1 reversed the inhibitory effect of HOTAIR knockdown on Wnt pathway.
Highly expressed HOTAIR promoted proliferation and migration of cardiac fibroblasts. HOTAIR remarkably upregulated fibrosis-related genes in CF cells. The mechanism of HOTAIR in regulating myocardial fibrosis might be related to the activation of Wnt signaling pathway through targeting URI1 expression.
探讨 HOX 转录反义 RNA（HOTAIR）是否通过促进心脏成纤维细胞（CFs）增殖和上调纤维化蛋白表达水平来调节心肌纤维化，其机制与激活 Wnt 信号通路有关。
采用实时定量聚合酶链反应（qRT-PCR）检测 Ang II 诱导的 CFs 中 HOTAIR 的表达水平。通过慢病毒转染过表达或敲低 HOTAIR 的表达。采用细胞计数试剂盒（CCK-8）检测、Transwell 检测和流式细胞术分别检测 HOTAIR 对细胞增殖、迁移和凋亡的影响。Western blot 和 qRT-PCR 实验检测纤维化相关基因和 Wnt 通路相关基因的表达。采用生物信息学分析预测 HOTAIR 的靶基因。通过 URI1 评估 HOTAIR 是否可以通过激活 Wnt 信号通路来调节细胞增殖和纤维化。
qRT-PCR 结果显示，Ang II 诱导的 CFs 中 HOTAIR 的表达明显高于对照组。CFs 中 HOTAIR 的过表达可促进细胞增殖和迁移，抑制细胞凋亡，并促进纤维化相关基因的表达。Western blot 结果表明，HOTAIR 可上调 URI1 的表达并激活 Wnt 信号通路。此外，挽救实验表明，URI1 的过表达逆转了 HOTAIR 敲低对 Wnt 通路的抑制作用。
高表达的 HOTAIR 促进了心脏成纤维细胞的增殖和迁移。HOTAIR 显著上调 CFs 中的纤维化相关基因。HOTAIR 调节心肌纤维化的机制可能与通过靶向 URI1 表达激活 Wnt 信号通路有关。
