Chen Lisha, Luo Cheng, Xu Yongcheng, Hu Jianjun, Chen Huixin
Department of Gastroenterology, Huizhou Municipal Central Hospital, Huizhou Central People's Hospital, Huizhou, China.
Anticancer Drugs. 2023 Apr 1;34(4):495-506. doi: 10.1097/CAD.0000000000001454. Epub 2022 Dec 2.
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.
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。