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环状RNA_0058063通过miR-377-3p/HOXA1轴调控食管癌的发展。

Circ_0058063 regulates the development of esophageal cancer through miR-377-3p/HOXA1 axis.

作者信息

Chen Lisha, Luo Cheng, Xu Yongcheng, Hu Jianjun, Chen Huixin

机构信息

Department of Gastroenterology, Huizhou Municipal Central Hospital, Huizhou Central People's Hospital, Huizhou, China.

出版信息

Anticancer Drugs. 2023 Apr 1;34(4):495-506. doi: 10.1097/CAD.0000000000001454. Epub 2022 Dec 2.

Abstract

Esophageal cancer is one of the deadliest cancers. Circular RNA (CircRNA) can be used as a tumor marker. Therefore, this provides an important idea for our research. Real-time quantitative PCR (RT-qPCR) was used to analyze the expression of circ_0058063, miR-377-3p and homeobox protein Hox-A1 (HOXA1), western blot was used to analyze the protein levels of HOXA1 and cyclinD1, B cell leukemia/lymphoma 2 associated X (Bax). Cell Counting Kit-8 (CCK-8) assay, colony formation assay and wound healing assay were used to analyze cell proliferation and migration; apoptosis was analyzed by flow cytometry. Dual-luciferase reporter assays were performed to analyze the luciferase activities. Transwell assay was used to analyze the cell invasion. A glycolysis metabolism assay was used to analyze cell glycolysis ability. Xenograft models were used to validate the effect of circ_0009035 in the growth of esophageal cancer in vivo . Circ_0009035 and HOXA1 were upregulated, while miR-377 was downregulated in esophageal cancer.. Circ_0058063 targeted miR-377-3p, and HOX4 was a target of miR-377-3p. Knockdown of circ_0058063 inhibited migration, invasion and proliferation and promoted apoptosis of esophageal cancer cells. MiR-377-3p inhibition or HOXA1 overexpression could restore the effect of si-circ_0058063 on esophageal cancer cells. Knockdown of circ_0058063 repressed the growth of esophageal cancer tumors in vivo. Our study found that circ_0058063 could regulate the expression of HOXA1 by targeting miR-377-3p, thereby affecting the progress of esophageal cancer.

摘要

食管癌是最致命的癌症之一。环状RNA(CircRNA)可作为肿瘤标志物。因此,这为我们的研究提供了重要思路。采用实时定量聚合酶链反应(RT-qPCR)分析环状RNA_0058063、微小RNA-377-3p(miR-377-3p)和同源框蛋白Hox-A1(HOXA1)的表达,采用蛋白质免疫印迹法分析HOXA1、细胞周期蛋白D1(cyclinD1)、B细胞淋巴瘤/白血病-2相关X蛋白(Bax)的蛋白水平。采用细胞计数试剂盒-8(CCK-8)法、集落形成试验和伤口愈合试验分析细胞增殖和迁移;通过流式细胞术分析细胞凋亡。进行双荧光素酶报告基因试验以分析荧光素酶活性。采用Transwell试验分析细胞侵袭。采用糖酵解代谢试验分析细胞糖酵解能力。采用异种移植模型在体内验证环状RNA_0009035对食管癌生长的影响。环状RNA_0009035和HOXA1在食管癌中上调,而miR-377下调。环状RNA_0058063靶向miR-377-3p,且HOX4是miR-377-3p的靶标。敲低环状RNA_0058063可抑制食管癌细胞的迁移、侵袭和增殖,并促进其凋亡。抑制miR-377-3p或过表达HOXA1可恢复si-环状RNA_0058063对食管癌细胞的作用。敲低环状RNA_0058063可抑制体内食管癌肿瘤的生长。我们的研究发现,环状RNA_0058063可通过靶向miR-377-3p调节HOXA1的表达,从而影响食管癌的进展。

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