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Hsa_circ_0005230 上调并通过调节 miR-1299/RHOT1 轴促进胃癌细胞侵袭和迁移。

Hsa_circ_0005230 is up-regulated and promotes gastric cancer cell invasion and migration via regulating the miR-1299/RHOT1 axis.

机构信息

Laboratory of Gastrointestinal Onco-Pathology, Cancer Institute & General Surgery Institute, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning Province, China.

出版信息

Bioengineered. 2022 Mar;13(3):5046-5063. doi: 10.1080/21655979.2022.2036514.

Abstract

Gastric cancer (GC) is one of the most common cancers in the world. Circular RNAs (circRNAs) are a class of non-coding RNAs that are widely expressed in eukaryotic cells. However, their role has been poorly understood in GC. This report aimed to explore the biological functions of hsa_circ_0005230 and its action mechanism in GC. This study validated that hsa_circ_0005230 was significantly up-regulated in 130 cases of GC tissues using qRT-PCR, and clinicopathological feature analysis revealed that its high expression was positively associated with histological grade, lymph node metastasis, TNM stages, and poor prognosis. In vitro, functional experiments showed that silencing hsa_circ_0005230 significantly decreased GC cell proliferation, invasion and migration capabilities. In addition, the major proteins of EMT (epithelial-mesenchymal transition) relevance have changed. In mechanism studies, bioinformatics analyses were used to predict the hsa_circ_0005230/miR-1299/RHOT1 axis and hsa_circ_0005230 may serve as a sponge for miR-1299 and indirectly regulate the expression of RHOT1. The regulated relationships between the molecules on the axis were verified using qRT-PCR and correlation analysis. Dual-luciferase reporter gene assay has been used to verify the binding site between miR-1299 and RHOT1. WB (Western blotting) and IHC (Immunohistochemical) were used to verify that RHOT1 may play the role of oncoprotein and affect the biological behavior of GC. Overall, hsa_circ_0005230 could enhance the EMT phenotype by promoting RHOT1 expression through sponging miR-1299, thus affecting the biological behavior of GC. Hsa_circ_0005230 can be easily identified as a potential diagnostic biomarker and assessment prognosis target for GC.

摘要

胃癌(GC)是世界上最常见的癌症之一。环状 RNA(circRNAs)是一类广泛存在于真核细胞中的非编码 RNA。然而,它们在 GC 中的作用还知之甚少。本报告旨在探讨 hsa_circ_0005230 的生物学功能及其在 GC 中的作用机制。本研究通过 qRT-PCR 验证了 hsa_circ_0005230 在 130 例 GC 组织中显著上调,临床病理特征分析表明其高表达与组织学分级、淋巴结转移、TNM 分期和预后不良呈正相关。在体外,功能实验表明,沉默 hsa_circ_0005230 显著降低 GC 细胞的增殖、侵袭和迁移能力。此外,EMT(上皮-间质转化)相关的主要蛋白发生变化。在机制研究中,利用生物信息学分析预测了 hsa_circ_0005230/miR-1299/RHOT1 轴,hsa_circ_0005230 可能作为 miR-1299 的海绵,并间接调节 RHOT1 的表达。通过 qRT-PCR 和相关性分析验证了轴上分子的调节关系。双荧光素酶报告基因实验验证了 miR-1299 和 RHOT1 之间的结合位点。WB(Western blotting)和 IHC(免疫组化)用于验证 RHOT1 可能作为癌蛋白发挥作用并影响 GC 的生物学行为。总的来说,hsa_circ_0005230 通过海绵吸附 miR-1299 促进 RHOT1 表达,从而增强 EMT 表型,影响 GC 的生物学行为。hsa_circ_0005230 可以很容易地被识别为 GC 的潜在诊断生物标志物和评估预后的靶标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6bb/8973856/35f87ea80e66/KBIE_A_2036514_UF0001_OC.jpg

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