钙激活中性蛋白酶1与口腔鳞状细胞癌迁移及侵袭的相关性  

Association of calcium-activated neutral protease 1 with the invasion and migration of oral squamous cell carcinoma

作　　者：谭丹 刘瑶 王忠朝 胡芸 胡妙龄 张梦雪 毛娅林 聂敏海 TAN Dan;LIU Yao;WANG ZhongChao;HU Yun;HU Miaoling;ZHANG Mengxue;MAO Yalin;NIE MinHai(Oral&Maxillofacial Reconstruction and Regeneration Laboratory,Southwest Medical University,Luzhou 646000,China;Department of periodontal mucosal diseases,Hostipal of stomatoloy,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学口腔颌面修复重建与再生实验室,泸州646000 [2]西南医科大学附属口腔医院牙周黏膜病科,泸州646000

出　　处：《西南医科大学学报》2022年第2期118-122,共5页Journal of Southwest Medical University

基　　金：四川省科技厅课题(2018JY041)。

摘　　要：目的在细胞和分子水平中探讨钙激活中性蛋白酶1(calcium activated neutral protease 1,CAPN1)与口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞侵袭、转移等行为是否存在相关性。方法选用永生化正常口腔黏膜上皮HOK细胞株作为对照组(正常组),2个不同来源的口腔鳞癌(OSCC)细胞株HSC3、CAL27作为实验组(肿瘤组),通过蛋白质印迹法(Western blot)探讨CAPN1在正常和肿瘤细胞中的表达情况。以OSCC细胞株中的两组HSC3、CAL27细胞为研究对象,将这两组细胞随机分为siRNA1-1组(将siRNA CAPN1-1转染到两组OSCC细胞中)、siRNA1-2组(将siRNA CAPN1-2转染到两组OSCC细胞中)及siRNA-neg组(将非特异性siRNA(siNC)转染到的两组OSCC细胞中)作为对照组。应用PCR法和Western blot检测转染后细胞内CAPN1mRNA和蛋白水平的变化。应用Transwell小室设计肿瘤细胞迁移、侵袭实验,检测转染后细胞迁移、侵袭能力的变化。结果Western blot表明CAPN1在肿瘤组中的表达高于正常组(P<0.05);通过PCR法和Western blot实验发现两组细胞HSC3、CAL27中,与siRNA-neg组相比,siRNA1-1组、siRNA1-2组中CAPN1mRNA的表达均明显受到抑制,差异均有统计学意义(P<0.05);通过Transwell小室实验发现siRNA1-1组与siRNA1-2组较siRNA-neg组迁移及侵袭能力均减弱,显微镜下穿膜数目均减少,差异均有统计学意义(P<0.01,P<0.05)。结论CAPN1可升高口腔鳞癌细胞迁移、侵袭能力,可能在口腔鳞癌的发生发展中发挥了重要作用。Objective To investigate the association of calcium-activated neutral protease 1(CAPN1)with the invasion and metastasis of oral squamous cell carcinoma(OSCC)cells.Methods The immortalized normal oral epithelial cell line HOK was selected as control(normal group),and the OSCC cell lines HSC3 and CAL27,with different origins,were selected for the experiment(tumor group).Western blot was used to measure the expression of CAPN1 in the normal group and the tumor group.Two groups of HSC3 and CAL27 cells in OSCC cell line were used as research subjects and were randomly divided into siRNA1-1 group(siRNA CAPN1-1 was transfected into the two groups of OSCC cells),siRNA1-2 group(siRNA CAPN1-2 was transfected into the two groups of OSCC cells),and siRNA-neg group(non-specific siRNA[siNC]was transfected into the two groups of OSCC cells as control group).PCR and Western blot were used to measure the changes in the mRNA and protein expression levels of CAPN1 in cells after transfection.Transwell chamber was used to design the migration and invasion experiment of tumor cells to observe the changes in the migration and invasion abilities of cells after transfection.Results Western blot showed that the expression of CAPN1 in the tumor group was significantly higher than that in the normal group(P<0.05).PCR and Western blot showed that for HSC3 and CAL27 cells,the siRNA1-1 group and the siRNA1-2 group had significant inhibition of the mRNA expression of CAPN1 compared with the siRNA-neg group(P<0.05).Transwell chamber assay showed that compared with the siRNA-neg group,the siRNA1-1 group and the siRNA1-2 group had significant reductions in migration and invasion abilities and the number of membrane penetration(P<0.01,P<0.05).Conclusion CAPN1 can increase the migration and invasion abilities of OSCC cells and may play an important role in the development and progression of OSCC.

关 键 词：中性钙蛋白酶 口腔鳞状细胞癌 RNA干扰 迁移 侵袭 相关性 

分 类 号：R78.59[医药卫生—口腔医学]