circ_0004390对口腔鳞癌细胞增殖、凋亡及顺铂敏感性的影响  

Regulatory effects of circ_0004390 on the proliferation and apoptosis of oral squamous cell carcinoma

作　　者：李伟 孟云 李玲 刘文成 吕志军 LI Wei;MENG Yun;LI Ling(Department of Stomatology,People's Hospital Affiliated to Shandong First Medical University,Shandong,Jinan 271199,China)

机构地区：[1]山东第一医科大学附属人民医院口腔科,济南市271199 [2]山东省济南市第二妇幼保健院口腔保健科

出　　处：《河北医药》2025年第3期368-374,共7页Hebei Medical Journal

基　　金：山东省优秀中青年科学家科研奖励基金(编号:BS2020SW407)。

摘　　要：目的探讨circ_0004390对口腔鳞癌细胞增殖、凋亡及顺铂敏感性的调控作用及机制。方法采用实时荧光定量PCR(quantitative real-time pcr,qRT-PCR)检测口腔鳞癌组织和细胞中circ_0004390的表达。采用Cell counting kit-8(CCK8)实验、克隆形成实验和EdU实验检测细胞增殖能力。采用流式细胞术检测细胞凋亡和线粒体膜电位。采用Western blot检测细胞中凋亡相关蛋白表达。采用CCK8实验检测不同浓度顺铂对细胞增殖的影响。通过在线数据库starBase分析circ_0004390的潜在靶miRNA及其下游靶基因。双荧光素酶报告基因实验验证基因的靶向关系。结果circ_0004390在口腔鳞癌组织和细胞中均呈高表达(P<0.05)。与si-NC组比较,si-circ_0004390组细胞的增殖活性和细胞克隆形成能力降低(P<0.05),细胞中EdU阳性率降低(P<0.05),细胞凋亡率升高(P<0.05),细胞线粒体膜电位降低(P<0.05),细胞中凋亡抑制蛋白Bcl2和Survivin表达明显减少(P<0.05),而促凋亡蛋白Bax、Cleaved-PARP和Cleaved-caspase3表达明显增多(P<0.05),且细胞对顺铂的敏感性增强(P<0.05)。过表达circ_0004390则表现出相反的作用。机制上,circ_0004390能够与miR-196b-5p靶向结合,且高迁移率蛋白A2(High mobility group AT-hook protein,HMGA2)是miR-196-5p的一个靶基因。结论沉默circ_0004390可抑制癌细胞的增殖,诱导细胞凋亡,增强细胞的顺铂敏感性。miR-196b-5p/HMGA2轴可能是circ_0004390的下游调控机制。Objective To investigate the regulatory effects of circ_0004390 on the proliferation,apoptosis and cisplatin sensitivity of oral squamous cell carcinoma(OSCC)and the underlying mechanism.Methods Expression level of circ_0004390 in OSCC tissue and cells was examined by quantitative real-time PCR(qRT-PCR).Cell proliferation was detected by cell counting kit-8(CCK-8)assay,colony formation assay and EdU(5-ethynyl-2'-deoxyuridine)assay.Apoptosis and mitochondrial membrane potential(MMP)were examined by flow cytometry.Apoptosis-associated proteins were measured by Western blot.Cholecystokinin-8(CCK-8)assay was further conducted to test the influence of cisplatin at various concentrations on cell proliferation.The potential target miRNAs and downstream of circ_0004390 were predicted by the starBase,and their target relationship was verified by dual-luciferase reporter assay.Results Circ_0004390 was highly expressed in OSCC(P<0.05).Transfection of si-circ_0004390 significantly reduced proliferative ability,colony formation ability,EdU-positive rate,MMP,and protein levels of anti-apoptotic Bcl-2 and Survivin,but elevated apoptotic rate,protein levels of pro-apoptotic Bax,cleaved-PAPR and cleaved-caspase-3,and sensitivity to cisplatin(P<0.05).Overexpression of circ_0004390 yielded the opposite results.Mechanically,circ_0004390 bound to miR-196b-5p,and HMGA2(high mobility group AT-hook protein)was the downstream target of miR-196b-5p.Conclusion Knockdown of circ_0004390 inhibits proliferation,induces apoptosis and enhances cisplatin sensitivity in OSCC by regulating the downstream miR-196b-5p/HMGA2 axis.

关 键 词：口腔鳞癌 circ_0004390 miR-196b-5p HMGA2 增殖 凋亡 

分 类 号：R739.8[医药卫生—肿瘤]