摘要
该文旨在探讨circNEIL3对口腔鳞癌细胞生物学行为的影响及其可能的作用机制。采用qRT-PCR法检测口腔鳞癌组织、癌旁组织、人口腔鳞癌细胞(CAL-27、SCC-25、SCC-9、HSC-3)以及正常口腔角质细胞HOK中circNEIL3、miR-218-5p的表达量;以CAL-27细胞为研究对象,si-circNEIL3、miR-218-5p mimics、si-NC、miR-NC分别转染至CAL-27细胞,si-circNEIL3和antimiR-NC、si-circNEIL3和anti-miR-218-5p分别共转染至CAL-27细胞;MTT法检测CAL-27细胞增殖情况;流式细胞术检测CAL-27细胞凋亡情况;Transwell检测CAL-27细胞侵袭和迁移情况。双荧光素酶报告实验检测circNEIL3与miR-218-5p的靶向关系;Western blot检测Bax、Bcl-2、caspase-3、cleaved-caspase-3蛋白表达量。与癌旁组织相比,口腔鳞癌组织中circNEIL3的表达量升高(P<0.01),miR-218-5p的表达量降低(P<0.01);与HOK细胞相比,人口腔鳞癌细胞(CAL-27、SCC-25、SCC-9、HSC-3)中circNEIL3的表达量升高(P<0.05),miR-218-5p的表达量降低(P<0.05)。分别与转染si-NC或miR-NC相比,转染si-circNEIL3或转染mi R-218-5p mimics增加细胞增殖抑制率、凋亡率和Bax、caspase-3、cleaved-caspase-3蛋白表达量,下调Bcl-2蛋白表达量,减少迁移、侵袭细胞数(P<0.01);circNEIL3可负向调控miR-218-5p的表达;与共转染si-circNEIL3和anti-miR-NC相比,共转染si-circNEIL3和anti-miR-218-5p后细胞增殖抑制率、细胞凋亡率和Bax、caspase-3、cleaved-caspase-3蛋白水平降低(P<0.01),迁移及侵袭细胞数增多(P<0.01),Bcl-2蛋白水平升高(P<0.01)。circNEIL3可通过靶向负调控miR-218-5p抑制口腔鳞癌细胞增殖、迁移及侵袭,并促进细胞凋亡。
This study aimed to investigate the effect of circNEIL3 targeting miR-218-5p on the biological behavior of oral squamous cell carcinoma cells.qRT-PCR was used to detect the expression of circNEIL3 and miR-218-5p in oral squamous cell carcinoma tissue,adjacent tissue and human oral squamous cell carcinoma cell(CAL-27,SCC-25,SCC-9,HSC-3)and normal oral keratinocyte HOK.Taking CAL-27 cells as the research object,si-circNEIL3,miR-218-5p mimics,si-NC,miR-NC were transfected into CAL-27 cells,si-circNEIL3 and anti-miR-NC,si-circNEIL3 and anti-miR-218-5p were co-transfected into CAL-27 cells.CAL-27 cell proliferation was detected by MTT assay;CAL-27 cell apoptosis was detected by flow cytometry;the invasion and migration of CAL-27 cells were detected by Transwell.Dual-luciferase reporter assay was used to detect the targeting relationship between circNEIL3 and miR-218-5p.Western blot was used to detect the expression of Bax,Bcl-2,caspase-3,and cleaved-caspase-3 protein.Compared with adjacent tissues,the expression of circNEIL3 in oral squamous cell carcinoma tissues was increased(P<0.01),while the expression of miR-218-5p was decreased(P<0.01);compared with HOK cells,the expression of circNEIL3 in human oral squamous cell carcinoma cells(CAL-27,SCC-25,SCC-9,HSC-3)was increased(P<0.05),and the expression of miR-218-5p was decreased(P<0.05).Compared with transfection of si-NC or miR-NC,transfection of si-circNEIL3 or miR-218-5p mimics increased cell proliferation inhibition rate,apoptosis rate,and Bax,caspase-3,and cleaved-caspase-3 protein levels(P<0.01),decreased Bcl-2 protein level(P<0.01),and reduced the number of migrating and invasive cells(P<0.01).circNEIL3 could negatively regulate the expression of miR-218-5p.Compared with co-transfection of si-circNEIL3 and anti-miR-NC,after co-transfection with si-circNEIL3 and anti-miR-218-5p,the cell proliferation inhibition rate,apoptosis rate and the protein levels of Bax,caspase-3,and cleaved-caspase-3 were reduced(P<0.01),while the number of migrating and invasive cells was increased(P<0.01),and the protein level of Bcl-2 was increased(P<0.01).circNEIL3 can inhibit the proliferation,migration and invasion of oral squamous cell carcinoma cells and promote cell apoptosis by targeting and negatively regulating miR-218-5p.
作者
刘月强
邢红艳
冯江洁
LIU Yueqiang;XING Hongyan;FENG Jiangjie(Department of Stomatology,Xinzhou Second People’s Hospital,Xinzhou 034100,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2023年第4期588-597,共10页
Chinese Journal of Cell Biology
