circ_0000326靶向miR-567对口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的调控效应  

Study on the molecular mechanism of circ_0000326 regulating the proliferation,invasion and migration of oral squamous cell carcinoma HSC3 cells by targeting miR-567

作　　者：刘文静 李梦琦 崔祥 汪俊兰[4] LIU Wen-jing;LI Meng-qi;CUI Xiang;WANG Jun-lan(Department of Stomatology,The Second People's Hospital of Hefei,Hefei Hospital Affiliated to Anhui Medical University,Hefei 230011,Anhui Province;Department of Stomatology,Affiliated Hospital of Jiangsu University,Zhenjiang 210031,Jiangsu Province;Kangbeijia Stomatological Hospital,Nanjing 210000,Jiangsu Province;Department of Stomatology,Jinling Hospital,Jinling Clinical College of Nanjing Medical University,Nanjing 210000,Jiangsu Province,China)

机构地区：[1]合肥市第二人民医院(安徽医科大学附属合肥医院)口腔科,安徽合肥230011 [2]江苏大学附属医院口腔科,江苏镇江210031 [3]南京康贝佳口腔医院,江苏南京210000 [4]南京医科大学附属金陵医院口腔科,江苏南京210000

出　　处：《上海口腔医学》2023年第6期590-596,共7页Shanghai Journal of Stomatology

摘　　要：目的:探讨circ_0000326调控口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的分子机制。方法:收集2020年3月—2021年6月安徽医科大学附属合肥医院收治的45例口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)患者的癌组织及癌旁组织标本,采用qRT-PCR检测circ_0000326、miR-567的表达量,CCK-8法、平板克隆形成实验、划痕实验和Transwell实验分别检测细胞增殖、克隆形成、迁移及侵袭能力,双荧光素酶报告实验检测circ_0000326与miR-567的靶向关系,Western印迹法检测E-cadherin、N-cadherin蛋白表达量。采用SPSS 21.0软件包对数据进行统计学分析。结果:circ_0000326在OSCC组织中的平均表达量为4.01±0.29,在癌旁组织的平均表达量为1.00±0.13;miR-567的表达量分别为0.28±0.03和1.00±0.10,差异具有统计学意义。与si-NC组相比,si-circ_0000326组细胞活力显著降低(P<0.05),细胞克隆形成数显著减少(P<0.05)。与si-NC组相比,si-circ_0000326组中侵袭细胞数、划痕愈合率显著降低(P<0.05),E-cadherin蛋白表达量显著升高(P<0.05),N-cadherin蛋白表达量显著降低(P<0.05)。Circ_0000326靶标miR-567。miR-567在pcDNA组的表达量为1.00±0.00,在pcDNA-circ_0000326组的表达量为0.44±0.04,在si-NC组的表达量为0.99±0.06,在si-circ_0000326组的表达量为2.92±0.25,差异具有统计学意义。与miR-NC组相比,miR-567组细胞活力显著降低、划痕愈合率及N-cadherin蛋白水平显著降低(P<0.05),细胞克隆形成数和侵袭细胞数显著减少(P<0.05),E-cadherin蛋白水平显著升高(P<0.05)。与si-circ_0000326+anti-miR-NC组相比,si-circ_0000326+anti-miR-567组细胞活力、划痕愈合率和N-cadherin蛋白水平显著升高(P<0.05),细胞克隆形成数和侵袭细胞数显著增多(P<0.05),E-cadherin蛋白水平显著降低(P<0.05)。结论:干扰circ_0000326表达可通过促进miR-567表达而减弱OSCC细胞增殖、克隆形成、迁移及侵袭能力。PURPOSE:To explore the molecular mechanism of circ_0000326 regulating proliferation,invasion and migration of oral squamous cell carcinoma HSC3 cells.METHODS:Cancerous tissue and adjacent tissue specimens of 45 patients with oral squamous cell carcinoma(OSCC)admitted to the Second People's Hospital of Hefei from March 2020 to June 2021 were collected.qRT-PCR was used to detect the expression levels of circ_0000326 and miR-567.CCK-8,plate clone formation test,scratch test and Transwell test were used to detect cell proliferation,clone formation,migration and invasion.Dual luciferase reporter experiment was used to detect the targeting relationship between circ_0000326 and miR-567.Western blot was used to quantify E-cadherin and N-cadherin protein.SPSS 21.0 software package was used for statistical analysis.RESULTS:circ_0000326 expression was 4.01±0.29 in OSCC and 1.00±0.13 in paracancerous tissues,while miR-567 expression was 0.28±0.03 and 1.00±0.10,respectively,with significant differences.Compared with the si-NC group,the cell viability and the number of cell clones in the si-circ_0000326 group were significantly decreased(P<0.05).Compared with the si-NC group,the number of invasive cells and scratch healing rate in the si-circ_0000326 group were significantly decreased(P<0.05),the expression level of E-cadherin protein was significantly increased(P<0.05),and the expression level of N-cadherin protein was significantly decreased(P<0.05).Additionally,circ_0000326 targeted miR-567.miR-567 expression was 1.00±0.00 in pcDNA group,0.44±0.04 in pcDNA-circ_0000326 group,0.99±0.06 in si-NC group,and 2.92±0.25 in si-circ_0000326 group with significant differences.Compared with miR-NC group,the cell viability,scratch healing rate,the number of cell clones and the number of invasive cells of miR-567 group were decreased,while E-cadherin protein level was increased(P<0.05).Compared with si-circ_0000326+anti-miR-NC group,the cell viability,scratch healing rate,N-cadherin protein level,the number of cell clones and th

关 键 词：口腔鳞状细胞癌 HSC3细胞系 circ_0000326 miR-567 细胞增殖 迁移 侵袭 

分 类 号：R739.8[医药卫生—肿瘤]