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长链非编码 RNA CCAT2 通过调控 miR-200b/VEGF 在骨肉瘤中发挥癌基因作用。

Long non-coding RNA CCAT2 acts as an oncogene in osteosarcoma through regulation of miR-200b/VEGF.

机构信息

a Department of Orthopedics, Qilu Hospital of Shandong University , Jinan , PR China.

b Department of Orthopedic Trauma, Yantai Affiliated Hospital of Binzhou Medical University , Yantai , PR China.

出版信息

Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2994-3003. doi: 10.1080/21691401.2019.1640229.

Abstract

Colon cancer-associated transcript 2 (CCAT2) is a new lncRNA, which is closely associated with risk of several cancers. The aim of this study was to explore the regulatory mechanism of CCAT2 in osteosarcoma (OSA). Cells were transfected with si-CCAT2, microRNA (miR)-200b inhibitor and the corresponding controls. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure the expression of CCAT2 and miR-200b in OSA tissues and cell lines. CCK8 and bromodeoxyuridine (BrdU) were conducted to examine cell proliferation. Apoptosis was detected by PI/FITC-Annexin V combining with flow cytometric analysis. Migration and invasion were respectively measured through transwell chambers assays. Western blot was used to examine expressions of relative proteins. CCAT2 was highly expressed and miR-200b was lowly expressed in OSA tissues and cell lines. Knockdown of CCAT2 suppressed cell proliferation, migration and invasion but induced apoptosis and up-regulation of miR-200b. miR-200b inhibitor weakened the effect of si-CCAT2 on cell progression and cell mobility. Besides, knockdown of CCAT2 blocked the PI3K/Akt and AMPK pathways through up-regulating miR-200b. The CCAT2/miR-200b/vascular endothelial growth factor (VEGF) axis plays important regulating effect in OSA through the PI3K/Akt and AMPK pathways.

摘要

结肠癌相关转录物 2(CCAT2)是一种新的长链非编码 RNA,与多种癌症的风险密切相关。本研究旨在探讨 CCAT2 在骨肉瘤(OSA)中的调控机制。将 si-CCAT2、microRNA(miR)-200b 抑制剂及其相应对照转染细胞。采用实时定量逆转录聚合酶链反应(qRT-PCR)检测 OSA 组织和细胞系中 CCAT2 和 miR-200b 的表达。CCK8 和溴脱氧尿苷(BrdU)检测细胞增殖。PI/FITC-Annexin V 结合流式细胞术检测细胞凋亡。Transwell 室测定分别检测细胞迁移和侵袭。Western blot 检测相关蛋白的表达。CCAT2 在 OSA 组织和细胞系中高表达,miR-200b 低表达。CCAT2 敲低抑制细胞增殖、迁移和侵袭,但诱导细胞凋亡和 miR-200b 上调。miR-200b 抑制剂减弱了 si-CCAT2 对细胞进展和细胞迁移能力的影响。此外,CCAT2 敲低通过上调 miR-200b 阻断了 PI3K/Akt 和 AMPK 通路。CCAT2/miR-200b/血管内皮生长因子(VEGF)轴通过 PI3K/Akt 和 AMPK 通路在 OSA 中发挥重要调节作用。

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