摘要
目的探讨circ_0006174靶向调控miR-876-3p对肺癌细胞恶性生物学行为的影响。方法体外培养A549细胞,随机分为si-circ_0006174组、si-NC组、si-circ_0006174+anti-miR-876-3p组、si-circ_0006174+anti-miRNC组,分别转染si-circ_0006174、si-NC、si-circ_0006174+anti-miR-876-3p、si-circ_0006174+anti-miR-NC,采用RTqPCR法检测circ_0006174、miR-876-3p表达。收集各组转染后细胞,采用CCK-8法检测细胞增殖情况,采用流式细胞术检测细胞凋亡情况,采用Transwell小室实验检测细胞侵袭和迁移情况,采用Western blotting法检测Bax、Bcl-2、E-cadherin、N-cadherin蛋白表达。通过Circular RNA Interactome在线软件预测circ_0006174与anti-miR-876-3p的靶向结合位点,并通过双荧光素酶报告基因实验验证二者的靶向调控关系。结果si-circ_0006174组circ_0006174相对表达量低于si-NC组,si-circ_0006174+anti-miR-876-3p组miR-876-3p相对表达量低于si-circ_0006174+anti-miRNC组(P均<0.05)。si-circ_0006174组细胞增殖抑制率和细胞凋亡率均高于si-NC组,而si-circ_0006174+anti-miR-876-3p组细胞增殖抑制率和细胞凋亡率均低于si-circ_0006174+anti-miR-NC组(P均<0.05)。si-circ_0006174组细胞侵袭和迁移能力均低于si-NC组,而si-circ_0006174+anti-miR-876-3p组细胞侵袭和迁移能力均高于si-circ_0006174+anti-miR-NC组(P均<0.05)。si-circ_0006174组Bax、E-cadherin蛋白相对表达量均高于si-NC组,Bcl-2、N-cadherin蛋白相对表达量均低于si-NC组(P均<0.05);si-circ_0006174+anti-miR-876-3p组Bax、E-cadherin蛋白相对表达量均低于si-circ_0006174+anti-miR-NC组,Bcl-2、N-cadherin蛋白相对表达量均高于si-circ_0006174+antimiR-NC组(P均<0.05)。经在线软件预测和双荧光素酶报告基因实验验证,circ_0006174能够靶向调控miR-876-3p。结论敲减circ_0006174可能通过靶向调控miR-876-3p抑制肺癌细胞恶性生物学行为。
Objective To explore the effect of circ006174 regulating miR-876-3 p on malignant biological behavior of lung cancer cells.Methods A549 cells were cultured in vitro,and randomly divided into the si-circ006174 group,si-NC group,si-circ006174+anti-miR-876-3 p group,and si-circ006174+anti-miR-NC group,which were transfected with si-circ006174,si-NC,si-circ006174+anti-miR-876-3 p,and si-circ006174+anti-miR-NC,respectively.The expression of circ006174 and miR-876-3 p was detected by RT-qPCR.The transfected cells in each group were collected,and the cell proliferation,apoptosis,and invasion and migration were detected by CCK-8 assay,flow cytometry,and Trans well chamber experiments,respectively.The protein expression levels of Bax,Bcl-2,E-cadherin,and N-cadherin were detected by Western blotting.The targeted binding sites between circ006174 and anti-miR-876-3 p were predicted by Circular RNA Interactome online software,and double luciferase reporter gene assay was used to verify the targeted regulation relationship between them.Results The relative expression of circ006174 was lower in the si-circ00617 group than in the si-NC group,while the relative expression of miR-876-3 p in the si-circ006174+antimiR-876-3 group was lower than that in the si-circ006174+anti-miR-NC group(both P<0.05).The cell proliferation inhibition and apoptosis rates in the si-circ006174 group were higher than those in the si-NC group,while cell proliferation inhibition and apoptosis rates of the si-circ006174+anti-miR-876-3 p group were lower than those of the si-circ006174+anti-miR-NC group(all P<0.05).The cell invasion and migration abilities of the si-circ006174 group were lower than those of the si-NC group,while the si-circ006174+anti-miR-876-3 p group had higher invasion and migration abilities than the si-circ006174+anti-miR-NC group(all P<0.05).The relative protein expression levels of Bax and E-cadherin in the si-circ006174 group were significantly higher than those in the si-NC group,and the relative expression levels of Bcl-2 and N-cadherin were significantly lower than those in the si-NC group(all P<0.05).The relative protein expression of Bax and E-cadherin in the si-circ006174+anti-miR-876-3 p group was significantly lower than that in the si-circ006174+anti-miR-NC group,while the relative expression of Bcl-2 and N-cadherin was significantly higher than that of the si-circ006174+anti-miR-NC group(all P<0.05).Circular RNA Interactome online software and experimental verification by double luciferase reporter assay revealed that circ006174 could targetedly regulate miR-876-3 p.Conclusion Knocking down circ006174 may inhibit the proliferation,invasion and migration of lung cancer cells and induce apoptosis by targeted regulation of miR-876-3 p.
作者
杨祎明
霍前伦
YANG Yiming;HUO qianlun(Department of Cardiothoracic Surgery,Lu'an Hospital Affiliated to Anhui Medical University,Lu'an 237005,China)
出处
《山东医药》
CAS
2022年第25期43-47,共5页
Shandong Medical Journal
