hsa_circ_0002141对口腔癌细胞生物学行为的调控作用  

Role of hsa_circ_0002141in regulating the biological behaviours of oral cancer cells

作　　者：宿伟鹏[1] 赵化荣[1] 李晨曦[2] 刘攀[1] 张洋[1] 龚忠诚[2] SU Wei-peng;ZHAO Hua-rong;LI Chen-xi;LIU Pan;ZHANG Yang;GONG Zhong-cheng(Cancer Center,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 830011,China;Department of Maxillofacial Oncology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 830011,China)

机构地区：[1]新疆医科大学第一附属医院肿瘤中心,新疆维吾尔自治区乌鲁木齐830011 [2]新疆医科大学第一附属医院颌面肿瘤外科,新疆维吾尔自治区乌鲁木齐830011

出　　处：《中华实用诊断与治疗杂志》2023年第1期28-32,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2021-6)。

摘　　要：目的筛选人口腔癌细胞系显著差异表达circRNA,探讨hsa_circ_0002141对人口腔癌细胞增殖、迁移、侵袭、凋亡及对口腔癌裸鼠肿瘤生长的影响。方法取人口腔癌细胞系HSC3、Sa3、SAS及人正常口腔角质形成细胞系HNOKs,采用高通量测序法检测circRNA表达,筛选显著差异表达的circRNA,取表达上调倍数最高的hsa_circ_0002141和高表达hsa_circ_0002141的SAS细胞进行后续实验。取SAS细胞分为对照组(正常培养)、空白转染组(转染sh-circNC慢病毒)、转染组(转染sh-circ_0002141慢病毒),转染2周,采用实时荧光定量PCR法检测3组细胞hsa_circ_0002141相对表达量,采用CCK-8法检测细胞增殖,采用Transwell小室实验检测细胞迁移及侵袭,采用流式细胞术测定细胞凋亡率。将6只裸鼠随机分为空白转染模型组和转染模型组各3只,分别接种转染sh-circNC慢病毒、sh-circ_0002141慢病毒的SAS细胞,于建模第3、6、9、12、15、18、21天测量肿瘤长径和短径,计算肿瘤体积;饲养21d处死2组裸鼠,测定肿瘤组织质量。结果HSC3、Sa3、SAS细胞与HNOKs细胞比较共发现152个显著差异表达circRNA,其中9个表达上调,143个表达下调,hsa_circ_0002141表达上调倍数最高。转染2周,转染组SAS细胞hsa_circ_0002141相对表达量(0.32±0.03)低于对照组(1.00±0.03)和空白转染组(1.02±0.07)(P<0.05)。转染后培养24、48、72、96h,转染组细胞增殖率[(87.24±8.49)%、(106.33±9.82)%、(130.29±13.06)%、(148.52±14.61)%]均低于对照组[(100.00±9.52)%、(136.17±13.23)%、(170.54±15.92)%、(190.75±18.99)%]和空白转染组[(100.29±9.56)%、(138.58±12.78)%、(171.48±16.51)%、(189.76±18.34)%](P<0.05)。转染后培养24h,转染组迁移细胞数[(120.05±11.54)个]、侵袭细胞数[(93.95±8.43)个]均少于对照组[(249.80±21.73)、(180.54±16.79)个]和空白转染组[(246.73±23.16)、(176.89±15.81)个](P<0.05)。转染2周,转染组细胞凋亡率[(22.68±2.15)%]高于对照组[(5.30±0.51)%]和�Objective To screen the significantly differentially expressed circRNAs in human oral cancer cell lines,and to investigate the influences of hsa_circ_0002141on the proliferation,migration,invasion and apoptosis of human oral cancer cells and the tumor growth in oral cancer nude mice.Methods The expressions of circRNA in human oral cancer cell lines HSC3,Sa3and SAS as well as human normal oral keratinocyte cell lines HNOKs were detected by high-throughput sequencing,the significantly differentially expressed circRNA was screened,and hsa_circ_0002141with the highest up-regulated expression and SAS cells with highly expressed hsa_circ_0002141 were selected for the subsequent experiment.The SAS cells were divided into control group(normal culture for 2weeks),blank transfection group(transfected with sh-circNC lentivirus for 2weeks)and transfection group(transfected with sh-circ_0002141 lentivirus for 2weeks).The relative expressions of hsa_circ_0002141were detected by real-time fluorescence quantitative PCR in three groups,the cell proliferation rates were determined by CCK-8assay,the numbers of migrated cells and invasive cells were determined by Transwell assay,and the apoptotic rates were determined by flow cytometry.Six nude mice were randomly and equally divided into blank transfection model group and transfection model group,inoculated with sh-circNC lentivirus and sh-circ_0002141lentivirus transfected SAS cells,respectively.The tumor volume was calculated after the long diameter(a)and short diameter(b)were measured by day 3,6,9,12,15,18and 21of modeling.After 21days of feeding,all nude mice were sacrificed and the tumor masses were measured.Results Comparing HSC3,Sa3 and SAS with HNOKs,152differentially expressed circRNAs were found,among which 9were up-regulated and 143 were down-regulated.With the highest up-regulated expression fold,hsa_circ_0002141was selected for the subsequent experiment.After 2-week transfection,the relative expression of hsa_circ_0002141of SAS cells was lower in transfection group(0.32

关 键 词：人口腔癌细胞 hsa_circRNA_0002141 增殖 迁移 侵袭 肿瘤生长 裸鼠 

分 类 号：R739.8[医药卫生—肿瘤]