Sun Suqing, Wang Peng, Ren Lijie, Wang Hongli, Zhan Yanli, Shan Shimin
Department of Anesthesia, Tianjin Fifth Central Hospital, Tianjin, People's Republic of China.
Onco Targets Ther. 2021 May 20;14:3319-3333. doi: 10.2147/OTT.S295552. eCollection 2021.
To explore the effect of SEV on colon cancer cells through circ-PI4KA.
The RNA level of circular RNA_0062389, microRNA-331-3p and LIM and SH3 protein 1 was determined by quantitative real-time polymerase chain reaction. Protein expression was detected by Western blot. Cell proliferation was investigated by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide, cell colony formation and 5-ethynyl-29-deoxyuridine assays. Cell apoptosis was demonstrated using Annexin V-fluorescein isothiocyanate/propidium iodide double staining assay. Cell migration and invasion were detected by transwell assay. The target relationship between miR-331-3p and circ-PI4KA or LASP1 was predicted by starBase v2.0 online database, and identified by a dual-luciferase reporter assay. The effects between SEV treatment and circ-PI4KA knockdown on tumor formation were presented by in vivo tumor formation assay.
Circ-PI4KA and LASP1 expressions were dramatically upregulated, while miR-331-3p was downregulated in colon cancer tissues and cells, respectively. SEV exposure significantly decreased the expression of circ-PI4KA and LASP1, but increased miR-331-3p expression. SEV inhibited cell proliferation, migration and invasion, and induced cell apoptosis by regulating circ-PI4KA. Furthermore, circ-PI4KA interacted with miR-331-3p, and miR-331-3p interacted with LASP1. SEV inhibited tumor growth by controlling circ-PI4KA in vivo.
Circ-PI4KA attenuated SEV-treated colon cancer cell malignancy by upregulating LASP1 through binding to miR-331-3p, which provided a new mechanism for studying surgery-mediated therapy of colon cancer.
通过circ-PI4KA探讨七氟醚(SEV)对结肠癌细胞的影响。
采用定量实时聚合酶链反应测定环状RNA_0062389、微小RNA-331-3p和LIM与SH3蛋白1的RNA水平。通过蛋白质免疫印迹法检测蛋白表达。采用3-(4,5)-二甲基噻唑-2,5-二苯基四氮唑溴盐、细胞集落形成和5-乙炔基-2'-脱氧尿苷检测法研究细胞增殖。使用膜联蛋白V-异硫氰酸荧光素/碘化丙啶双染法证明细胞凋亡。通过Transwell检测法检测细胞迁移和侵袭。通过starBase v2.0在线数据库预测miR-331-3p与circ-PI4KA或LASP1之间的靶标关系,并通过双荧光素酶报告基因检测法进行鉴定。通过体内肿瘤形成试验呈现SEV处理和circ-PI4KA敲低对肿瘤形成的影响。
circ-PI4KA和LASP1的表达在结肠癌组织和细胞中分别显著上调,而miR-331-3p下调。SEV暴露显著降低circ-PI4KA和LASP1的表达,但增加miR-331-3p的表达。SEV通过调节circ-PI4KA抑制细胞增殖、迁移和侵袭,并诱导细胞凋亡。此外,circ-PI4KA与miR-331-3p相互作用,且miR-331-3p与LASP1相互作用。SEV在体内通过控制circ-PI4KA抑制肿瘤生长。
circ-PI4KA通过与miR-331-3p结合上调LASP1,从而减弱SEV处理的结肠癌细胞的恶性程度,这为研究手术介导的结肠癌治疗提供了一种新机制。