Hsa_circ_0072765 敲低通过 miR-197-3p/TRPV3 轴抑制转化生长因子-β (TGF-β)诱导的肝星状细胞 (HSCs)的增殖、激活和迁移。
Hsa_circ_0072765 knockdown inhibits proliferation, activation and migration in transforming growth factor-beta (TGF-β)-induced hepatic stellate cells (HSCs) by the miR-197-3p/TRPV3 axis.
机构信息
Department of Gastroenterology, Shaanxi Provincial People's Hospital, Xi'an City, Shaanxi, China.
出版信息
Histol Histopathol. 2023 Nov;38(11):1295-1306. doi: 10.14670/HH-18-586. Epub 2023 Jan 17.
BACKGROUND
Circular RNAs (circRNAs) participate in the progression of diverse human diseases. However, the effects of circRNAs on liver fibrosis are limited. In this study, we aimed to investigate the functions of hsa_circ_0072765 in liver fibrosis.
METHODS
Transforming growth factor-beta (TGF-β)-treated hepatic stellate cells (HSCs) were used as the cell model of liver fibrosis. Quantitative real-time polymerase chain reaction (qRT-PCR) or western blot was performed to determine the expression of hsa_circ_0072765, microRNA-197-3p (miR-197-3p) and transient receptor potential cation channel subfamily V member 3 (TRPV3). 5'-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry analysis and wound-healing assay were conducted to evaluate cell proliferation, cell cycle and migration. HSC activation was assessed by determining the expression of alpha-smooth muscle actin (α-SMA) and type I collagen alpha 1 (Col1A1). Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) were manipulated to analyze the relationship of hsa_circ_0072765, miR-197-3p and TRPV3. The exosome morphology was observed under transmission electron microscopy (TEM).
RESULTS
Hsa_circ_0072765 level was increased in TGF-β-induced HSCs. Hsa_circ_0072765 knockdown inhibited cell proliferation, cell cycle, activation and migration in TGF-β-induced HSCs. Hsa_circ_0072765 sponged miR-197-3p and negatively regulated miR-197-3p expression. MiR-197-3p inhibition reversed the effects of hsa_circ_0072765 knockdown on TGF-β-induced HSC proliferation, cell cycle, activation and migration. In addition, TRPV3 was the target gene of miR-197-3p and miR-197-3p overexpression inhibited TGF-β-treated HSC proliferation, cell cycle, activation and migration by targeting TRPV3. Besides, we found that exosomal hsa_circ_0072765 was increased in TGF-β-treated HSCs.
CONCLUSION
Hsa_circ_0072765 promoted the progression of TGF-β-treated HSCs by decoying miR-197-3p and upregulating TRPV3.
背景
环状 RNA（circRNAs）参与多种人类疾病的进展。然而，circRNAs 对肝纤维化的影响有限。在这项研究中，我们旨在研究 hsa_circ_0072765 在肝纤维化中的功能。
方法
转化生长因子-β（TGF-β）处理的肝星状细胞（HSCs）用作肝纤维化的细胞模型。实时定量聚合酶链反应（qRT-PCR）或 Western blot 用于测定 hsa_circ_0072765、微小 RNA-197-3p（miR-197-3p）和瞬时受体电位阳离子通道亚家族 V 成员 3（TRPV3）的表达。5'-乙炔基-2'-脱氧尿苷（EdU）测定、流式细胞术分析和划痕愈合试验用于评估细胞增殖、细胞周期和迁移。通过测定α-平滑肌肌动蛋白（α-SMA）和 I 型胶原α1（Col1A1）的表达来评估 HSC 激活。双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）用于分析 hsa_circ_0072765、miR-197-3p 和 TRPV3 之间的关系。透射电子显微镜（TEM）观察外体形态。
结果
TGF-β诱导的 HSCs 中 hsa_circ_0072765 水平升高。hsa_circ_0072765 敲低抑制 TGF-β 诱导的 HSCs 中的细胞增殖、细胞周期、激活和迁移。hsa_circ_0072765 海绵 miR-197-3p 并负调控 miR-197-3p 表达。miR-197-3p 抑制逆转了 hsa_circ_0072765 敲低对 TGF-β 诱导的 HSC 增殖、细胞周期、激活和迁移的影响。此外，TRPV3 是 miR-197-3p 的靶基因，miR-197-3p 过表达通过靶向 TRPV3 抑制 TGF-β 处理的 HSC 增殖、细胞周期、激活和迁移。此外，我们发现 TGF-β 处理的 HSCs 中 exosomal hsa_circ_0072765 增加。
结论
hsa_circ_0072765 通过诱饵 miR-197-3p 和上调 TRPV3 促进 TGF-β 处理的 HSCs 的进展。
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