Wang Jingpeng, Li Shuyuan, Zhang Gaofeng, Han Huihua
Department of Anaesthesiology, The Chengyang People's Hospital, No.76 Zhengyang Road, Chengyang District, Qingdao, 266109, Shandong Province, China.
Fever Clinic, The Chengyang People's Hospital, Qingdao, Shandong Province, China.
J Biol Res (Thessalon). 2021 Jun 28;28(1):14. doi: 10.1186/s40709-021-00145-6.
Sevoflurane (Sev), a commonly used volatile anesthetic, has been reported to inhibit the process of colorectal cancer (CRC). Circular RNAs (circRNAs) are revealed to participate in the pathogenesis of CRC. This study aims to reveal the mechanism of hsa_circ_0000231 in Sev-mediated CRC progression.
The expression of hsa_circ_0000231 and microRNA-622 (miR-622) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein level was determined by western blot analysis. Cell proliferation was investigated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), cell colony formation and DNA content quantitation assays. Cell apoptosis was detected by Annexin V-fluorescein isothiocyanate and propidium iodide double staining and caspase 3 activity assays. Cell migration and invasion were investigated by wound-healing and transwell invasion assays, respectively. The putative relationship between hsa_circ_0000231 and miR-622 was predicted by circular RNA Interactome online database, and identified by dual-luciferase reporter and RNA immunoprecipitation assays. The impacts of hsa_circ_0000231 on Sev-mediated tumor formation in vivo were presented by in vivo assay.
Hsa_circ_0000231 expression was upregulated, while miR-622 was downregulated in CRC tissues and cells compared with control groups. Sev treatment decreased hsa_circ_0000231 expression, but increased miR-622 expression in CRC cells. Sev treatment suppressed cell proliferation, migration and invasion, and induced cell apoptosis. Hsa_circ_0000231 overexpression restored Sev-mediated CRC progression in vitro. Additionally, hsa_circ_0000231 acted as a sponge of miR-622, and miR-622 inhibitors reversed the impacts of hsa_circ_0000231 silencing on CRC process. Furthermore, Sev treatment inhibited tumor growth by regulating hsa_circ_0000231 in vivo.
Hsa_circ_0000231 attenuated Sev-aroused repression impacts on CRC development by sponging miR-622. This findings may provide an appropriate anesthetic protocol for CRC sufferers undergoing surgery.
七氟醚(Sev)是一种常用的挥发性麻醉剂,据报道可抑制结直肠癌(CRC)的进程。环状RNA(circRNAs)被发现参与CRC的发病机制。本研究旨在揭示hsa_circ_0000231在Sev介导的CRC进展中的作用机制。
采用定量实时聚合酶链反应(qRT-PCR)检测hsa_circ_0000231和微小RNA-622(miR-622)的表达。通过蛋白质印迹分析测定蛋白质水平。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)、细胞集落形成和DNA含量定量分析研究细胞增殖。通过膜联蛋白V-异硫氰酸荧光素和碘化丙啶双重染色以及半胱天冬酶3活性测定检测细胞凋亡。分别采用伤口愈合和Transwell侵袭试验研究细胞迁移和侵袭。通过环状RNA相互作用组在线数据库预测hsa_circ_0000231与miR-622之间的潜在关系,并通过双荧光素酶报告基因和RNA免疫沉淀试验进行鉴定。通过体内试验呈现hsa_circ_0000231对Sev介导的体内肿瘤形成的影响。
与对照组相比,CRC组织和细胞中hsa_circ_0000231表达上调,而miR-622表达下调。Sev处理降低了CRC细胞中hsa_circ_0000231的表达,但增加了miR-622的表达。Sev处理抑制细胞增殖、迁移和侵袭,并诱导细胞凋亡。hsa_circ_0000231过表达恢复了Sev介导的体外CRC进展。此外,hsa_circ_0000231作为miR-622的海绵,miR-622抑制剂逆转了hsa_circ_0000231沉默对CRC进程的影响。此外,Sev处理通过在体内调节hsa_circ_0000231抑制肿瘤生长。
hsa_circ_0000231通过海绵吸附miR-622减弱了Sev对CRC发展的抑制作用。这一发现可能为接受手术的CRC患者提供合适的麻醉方案。