靶向沉默热休克蛋白27对口腔鳞状细胞癌CAL27细胞侵袭和迁移的作用及其机制  

Effects of targeted silencing of heat shock protein 27 on invasion and migration of oral squamous cell carcinoma CAL27 cells and their mechanisms

作　　者：曹娟 李玮柏 郭秀 李波 董春玲[2] CAO Juan;LI Weibo;GUO Xiu;LI Bo;DONG Chunling(Experimental Teaching Center,Stomatology Hospital,Jilin University,Changchun 130021,China;Department of Respiratory Medicine,Second Hospital,Jilin University,Changchun 130041,China;Experimental Teaching Center,Affiliated Stomatological Hospital,School of Stomatology,China Medical University,Key Laboratory of Oral Diseases of Liaoning Province,Shenyang 110002,China)

机构地区：[1]吉林大学口腔医院实验教学中心,吉林长春130021 [2]吉林大学第二医院呼吸内科,吉林长春130041 [3]中国医科大学口腔医学院·附属口腔医院实验教学中心辽宁省口腔疾病重点实验室,辽宁沈阳110002

出　　处：《吉林大学学报（医学版）》2021年第4期971-977,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金项目(20200201329JC);吉林省科技厅国际科技合作项目(20170414057GH);吉林省教育厅“十三五”科学技术研究规划项目(JJKH20201110KJ);吉林省财政厅优秀中青年骨干人才项目(JCSZ2019378-14);吉林省财政厅卫生专项项目(2020SCZT023,3D5177713429);吉林省卫健委青年科技骨干培养计划项目(2016Q020,2015Z007);吉林大学白求恩计划科研项目(2018B27)。

摘　　要：目的:探讨热休克蛋白27(HSP27)对口腔鳞状细胞癌(OSCC)CAL27细胞侵袭和迁移的影响,并阐明其作用机制。方法:CAL27细胞分为对照组[转染阴性对照小干扰RNA(NC-siRNA)]和实验组[转染HSP27小干扰RNA(HSP27-siRNA)],实时荧光定量PCR(RT-qPCR)和Western blotting法检测2组CAL27细胞siRNA干扰效率,细胞划痕实验检测2组CAL27细胞划痕愈合率,Transwell小室实验检测2组CAL27细胞侵袭和迁移细胞数,RT-qPCR和Western blotting法检测2组CAL27细胞中上皮-间质转化(EMT)标记物N-钙黏蛋白(N-cadherin)mRNA和蛋白表达水平。结果:与对照组比较,实验组CLA27细胞中HSP27 mRNA和蛋白表达水平明显降低(P<0.01)。细胞划痕实验,与对照组比较,实验组CAL27细胞划痕愈合率明显降低(P<0.01)。Transwell小室实验,与对照组比较,实验组CAL27细胞中迁移和侵袭细胞数明显减少(P<0.01)。与对照组比较,实验组CAL27细胞中N-cadherin mRNA和蛋白表达水平明显降低(P<0.05)。结论:靶向沉默HSP27可以导致OSCC CAL27细胞迁移和侵袭能力降低,可能与抑制EMT有关。Objective:To investigate the effects of heat shock protein 27(HSP27)on the invasion and migration of oral squamous cell carcinoma(OSCC) CAL27 cells,and to clarify their mechanisms.Methods:The CAL27 cells were divided into control group[transfected with negative control interference RNA(NC-siRNA)]and experiment group[transfected with HSP27 small interference RNA(HSPsiRNA)]. Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the siRNA interference efficiency in the CAL27 cells. Cell scratch assay was used to detect the scratch healing rates of CAL27 cells. Transwell chamber experiment was used to detect the number of invasion and migration CAL27 cells in two groups. RT-qPCR and Western blotting methods were used to detect the mRNA and protein expression levels of epithelial mesenchymal transformation(EMT)marker Ncadherin in the CAL27 cells in two groups. Results:Compared with control group,the expression levels of HSP27 mRNA and protein in the CLA27 cells in experiment group were significantly decreased(P<0. 01). The results of cell scratch experiment showed that compared with control group,the scratch healing rate of the CAL27 cells in experiment group was significantly decreased(P<0. 01). The results of Transwell chamber experiment showed that compared with control group,the number of invasion and migration cells in experiment group was significantly reduced(P<0. 01). Compared with control group,the expression levels of N-cadherin mRNA and protein in the CAL27 cells in expermient group were significantly reduced(P<0. 05). Conclusion:Targeted silencing of HSP27 can lead to the reduced migration and invasion abilities of the OSCC CAL27 cells,which may be related to the inhibition of EMT.

关 键 词：热休克蛋白27 口腔鳞状细胞癌 细胞侵袭 细胞迁移 上皮-间质转化 

分 类 号：R739.8[医药卫生—肿瘤]