m6A 修饰的 circ_0124554 通过 miR-1184/LASP1 通路促进结直肠癌的进展和放射抵抗。
m6A-modified circ_0124554 promotes colorectal cancer progression and radioresistance through the miR-1184/LASP1 pathway.
机构信息
Department of Oncology, People's Hospital of Ningxiang, Changsha City, Hunan Province, China.
Department of Oncology, The Affiliated Changsha Central Hospital, Hengyang Medical School, University of South China, Changsha City, Hunan Province, China.
出版信息
Pathol Res Pract. 2024 Jan;253:154950. doi: 10.1016/j.prp.2023.154950. Epub 2023 Nov 14.
BACKGROUND
Circular RNAs (circRNAs) are believed to regulate the progression of various cancers including colorectal cancer (CRC). However, the role and mechanism of circ_0124554 in regulating the sensitivity of CRC to radiation remain unknown.
METHODS
The RNA levels of circ_0124554, LIM and SH3 protein 1 (LASP1), and methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit (METTL3) were detected by quantitative real-time polymerase chain reaction. Protein expression was checked by western blot. Cell proliferation, apoptosis, migration, and invasion were investigated by 5-Ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry analysis, and transwell assay, respectively. The sensitivity of CRC cells to radiation was analyzed by cell colony formation assay. Xenograft mouse model assay was conducted to disclose the role of circ_0001023 in the sensitivity of tumors to radiation in vivo. The binding relationships among circ_0124554, miR-1184 and LASP1 were confirmed by a dual-luciferase reporter assay. m6A RNA immunoprecipitation assay was performed to identify the association of METTL3 with circ_0124554.
RESULTS
Circ_0124554 expression was upregulated in CRC tissues and cells in comparison with normal colorectal tissues and cells. Circ_0124554 knockdown inhibited proliferation, migration and invasion and promoted apoptosis and radiosensitivity of CRC cells. Moreover, circ_0124554 depletion inhibited tumor formation and improved radiosensitivity in vivo. MiR-1184 was identified as a target miRNA of circ_0124554 and targeted LASP1. Additionally, LASP1 overexpression rescued circ_0124554 knockdown-mediated effects in CRC cells. METTL3 mediated m6A methylation of circ_0124554. Further, circ_0124554 overexpression attenuated METTL3 depletion-induced effects in CRC cells.
CONCLUSION
m6A-modified circ_0124554 promoted CRC progression and radioresistance by inducing LASP1 expression through interaction with miR-1184.
背景
环状 RNA（circRNAs）被认为可以调节多种癌症的进展，包括结直肠癌（CRC）。然而，circ_0124554 调节 CRC 对辐射敏感性的作用和机制尚不清楚。
方法
通过实时定量聚合酶链反应检测 circ_0124554、LIM 和 SH3 蛋白 1（LASP1）以及甲基转移酶 3、N6-腺苷-甲基转移酶复合物催化亚基（METTL3）的 RNA 水平。通过蛋白质印迹检测蛋白表达。通过 5-乙炔基-2'-脱氧尿苷（EdU）检测、流式细胞术分析和 Transwell 检测分别检测细胞增殖、凋亡、迁移和侵袭。通过细胞集落形成实验分析 CRC 细胞对辐射的敏感性。通过异种移植小鼠模型实验在体内揭示 circ_0001023 在肿瘤对辐射敏感性中的作用。通过双荧光素酶报告基因实验证实 circ_0124554 与 miR-1184 和 LASP1 之间的结合关系。通过 m6A RNA 免疫沉淀实验鉴定 METTL3 与 circ_0124554 的关联。
结果
与正常结直肠组织和细胞相比，CRC 组织和细胞中 circ_0124554 的表达上调。circ_0124554 敲低抑制 CRC 细胞的增殖、迁移和侵袭，促进凋亡和放射敏感性。此外，circ_0124554 耗竭抑制体内肿瘤形成并提高放射敏感性。MiR-1184 被鉴定为 circ_0124554 的靶 miRNA，并靶向 LASP1。此外，LASP1 的过表达挽救了 CRC 细胞中 circ_0124554 敲低介导的效应。METTL3 介导 circ_0124554 的 m6A 甲基化。此外，circ_0124554 的过表达减弱了 CRC 细胞中 METTL3 耗竭诱导的效应。
结论
m6A 修饰的 circ_0124554 通过与 miR-1184 相互作用诱导 LASP1 表达，促进 CRC 进展和放射抗性。
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