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Circ_0001273 下调通过靶向 miR-622/SLC1A5 信号轴抑制食管癌细胞的生长、迁移和谷氨酰胺代谢。

Circ_0001273 downregulation inhibits the growth, migration and glutamine metabolism of esophageal cancer cells via targeting the miR-622/SLC1A5 signaling axis.

机构信息

Department of Thoracic Surgery, Gaozhou People's Hospital, Gaozhou City, Guangdong Province, China.

出版信息

Thorac Cancer. 2022 Jun;13(12):1795-1805. doi: 10.1111/1759-7714.14458. Epub 2022 May 13.

DOI:10.1111/1759-7714.14458
PMID:35567340
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9200876/
Abstract

BACKGROUND

Esophageal cancer is a relatively rare cancer. However, its death rate is not to be taken lightly. Accumulating evidence indicates circular RNA (circRNA) is implicated in cancer development. The objective of this study was to unveil the role of circ_0001273 in esophageal cancer (EC).

METHODS

For expression analysis of circ_0001273, miR-622 and solute carrier family 1 member 5 (SLC1A5), quantitative real-time PCR (qPCR) and Western blot were conducted. Cell proliferation was evaluated by cell counting kit-8 (CCK-8), EdU and colony formation assays. Cell apoptosis and cell migration were investigated using flow cytometry assay and wound healing assay. Glutamine metabolism was assessed by glutamine consumption and glutamate production using matched kits. The predicted binding relationship between miR-622 and circ_0001273 or SLC1A5 was validated by dual-luciferase reporter assay. An in vivo xenograft model was established to determine the role of circ_0001273 on tumor growth.

RESULTS

Circ_0001273 was upregulated in EC tumor tissues and cells. Knockdown of circ_0001273 repressed EC cell proliferation, migration, epithelial-mesenchymal transition (EMT) and glutamine metabolism. Circ_0001273 knockdown also blocked tumor development in animal models. MiR-622 was targeted by circ_0001273, and its inhibition reversed the functional effects of circ_0001273 knockdown. SLC1A5 was a target gene of miR-622, and circ_0001273 targeted miR-622 to positively regulate SLC1A5 expression. The inhibitory effects of miR-622 enrichment on EC cell proliferation, migration, EMT and glutamine metabolism were recovered by SLC1A5 overexpression.

CONCLUSION

Circ_0001273 high expression contributed to EC progression via modulating the miR-622/SLC1A5 signaling axis.

摘要

背景

食管癌是一种相对罕见的癌症，但它的死亡率不容忽视。越来越多的证据表明，环状 RNA（circRNA）与癌症的发展有关。本研究的目的是揭示环状 RNA（circ_0001273）在食管癌（EC）中的作用。

方法

为了分析 circ_0001273、miR-622 和溶质载体家族 1 成员 5（SLC1A5）的表达，进行了定量实时 PCR（qPCR）和 Western blot 检测。通过细胞计数试剂盒-8（CCK-8）、EdU 和集落形成实验评估细胞增殖。通过流式细胞术检测和划痕愈合实验检测细胞凋亡和细胞迁移。使用匹配的试剂盒评估谷氨酰胺代谢，包括谷氨酰胺消耗和谷氨酸产生。通过双荧光素酶报告基因实验验证 miR-622 与 circ_0001273 或 SLC1A5 的预测结合关系。建立体内异种移植模型以确定 circ_0001273 对肿瘤生长的作用。

结果

circ_0001273 在 EC 肿瘤组织和细胞中上调。circ_0001273 敲低抑制 EC 细胞增殖、迁移、上皮-间充质转化（EMT）和谷氨酰胺代谢。circ_0001273 敲低也阻止了动物模型中的肿瘤发展。miR-622 是 circ_0001273 的靶基因，其抑制作用逆转了 circ_0001273 敲低的功能效应。SLC1A5 是 miR-622 的靶基因，circ_0001273 通过靶向 miR-622 正向调节 SLC1A5 表达。SLC1A5 过表达恢复了 miR-622 富集对 EC 细胞增殖、迁移、EMT 和谷氨酰胺代谢的抑制作用。

结论

circ_0001273 的高表达通过调节 miR-622/SLC1A5 信号轴促进 EC 的进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/62321eb95c36/TCA-13-1795-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/303625dfff1b/TCA-13-1795-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/796b75f5a66f/TCA-13-1795-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/6e5fc6d9a494/TCA-13-1795-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/d6f22caf303b/TCA-13-1795-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/065c12d1c674/TCA-13-1795-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/2571aa953780/TCA-13-1795-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/62321eb95c36/TCA-13-1795-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/303625dfff1b/TCA-13-1795-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/796b75f5a66f/TCA-13-1795-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/6e5fc6d9a494/TCA-13-1795-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/d6f22caf303b/TCA-13-1795-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/065c12d1c674/TCA-13-1795-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/2571aa953780/TCA-13-1795-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a1/9200876/62321eb95c36/TCA-13-1795-g001.jpg

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