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长链非编码 RNA DANCR 通过调节 Twist1 海绵吸附 miR-345-5p 影响胆管癌细胞的生长、EMT 和血管生成。

LncRNA DANCR affected cell growth, EMT and angiogenesis by sponging miR-345-5p through modulating Twist1 in cholangiocarcinoma.

机构信息

Department of Abdominal Surgery, Linyi Cancer Hospital, Linyi, Shandong, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Mar;24(5):2321-2334. doi: 10.26355/eurrev_202003_20498.

Abstract

OBJECTIVE

LncRNA DANCR has been reported to play an important role in various cancers. Therefore, this study aimed at exploring the function and regulatory mechanism of DANCR in Cholangiocarcinoma (CCA).

PATIENTS AND METHODS

qRT-PCR was used to measure the expression of DANCR, miR-345-5p in tissues and cells. Western blot was applied to measure the protein expression of Twist, N-cadherin, Vimentin, E-cadherin, VEGF-A, VEGF-C, PCNA and C-caspase 3. The relationship between DANCR and miR-345-5p was determined by luciferase reporter assay. MTT assay and flow cytometry were used to assess cell proliferation and apoptosis, respectively. Transwell assay was performed to detect cell invasion and migration.

RESULTS

We found that the expression of DANCR was significantly induced in CCA tissues and cells. Inhibition of DANCR remarkably suppressed CCA cell proliferation, migration, invasion, EMT and angiogenesis as well as induced cell apoptosis in vitro and in vivo. Luciferase reporter assay determined that DANCR directly targeted miR-345-5p and Twist1 was a target mRNA of miR-345-5p. Otherwise, miR-345-5p down-expression partially reversed the effect induced by the suppression of DANCR in CCA. Moreover, the suppressive effects of high miR-345-5p expression on CCA cells were reversed by improving Twist1 expression.

CONCLUSIONS

In this study, we verified that LncRNA DANCR affected cell proliferation, migration, invasion, angiogenesis, epithelial-mesenchymal transition (EMT) and induced apoptosis through modulating miR-345-5p/Twist1 axis in Cholangiocarcinoma.

摘要

目的

长链非编码 RNA DANCR 已被报道在多种癌症中发挥重要作用。因此,本研究旨在探讨 DANCR 在胆管癌(CCA)中的功能和调控机制。

患者和方法

qRT-PCR 用于测量组织和细胞中 DANCR 和 miR-345-5p 的表达。Western blot 用于测量 Twist、N-钙黏蛋白、波形蛋白、E-钙黏蛋白、VEGF-A、VEGF-C、PCNA 和 C-caspase 3 的蛋白表达。通过荧光素酶报告基因检测确定 DANCR 和 miR-345-5p 之间的关系。MTT assay 和流式细胞术分别用于评估细胞增殖和凋亡。Transwell assay 用于检测细胞侵袭和迁移。

结果

我们发现 DANCR 在 CCA 组织和细胞中的表达明显上调。抑制 DANCR 可显著抑制 CCA 细胞的增殖、迁移、侵袭、上皮间质转化(EMT)和血管生成,并在体内外诱导细胞凋亡。荧光素酶报告基因检测确定 DANCR 直接靶向 miR-345-5p,Twist1 是 miR-345-5p 的靶 mRNA。此外,miR-345-5p 的下调表达部分逆转了 DANCR 抑制对 CCA 产生的影响。此外,高表达 miR-345-5p 对 CCA 细胞的抑制作用通过提高 Twist1 表达而逆转。

结论

在本研究中,我们通过调节 miR-345-5p/Twist1 轴证实,长链非编码 RNA DANCR 通过调节 miR-345-5p/Twist1 轴影响胆管癌细胞的增殖、迁移、侵袭、血管生成、上皮间质转化(EMT)和诱导凋亡。

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