摘要
目的检测核苷酸结合寡聚化结构域样受体蛋白1(NLRP1)在口腔鳞状细胞癌(OSCC)组织中的表达及其对OSCC细胞系CAL-27增殖、迁移和侵袭的影响。方法收集2021年4月至2022年8月贵州省人民医院口腔颌面外科收治的手术患者中原发性OSCC肿瘤组织样本及癌旁正常组织样本各40例,采用反转录荧光定量聚合酶链反应(RT-qPCR)检测40例OSCC组织及对应癌旁正常组织中NLRP1信使RNA(mRNA)的表达情况,结合患者临床病理参数进行相关性分析。利用RT-qPCR检测NLRP1在OSCC细胞系CAL-27和正常口腔黏膜上皮细胞(OMEC)中的表达。通过瞬时转染方式将pcDNA-NLRP1转染至OSCC细胞系CAL-27中构建体外NLRP1过表达细胞模型,转染pcDNA-NC作为阴性对照组,利用RT-qPCR和Western blot检测细胞转染效率,采用细胞计数试剂-8法检测过表达NLRP1对CAL-27细胞增殖的影响。利用划痕愈合实验和Transwell小室实验检测CAL-27细胞迁移和侵袭能力。结果OSCC组织中NLRP1 mRNA表达显著低于癌旁组织[0.49(0.13,0.99)比1.02(1.00,1.03)](Z=4.224,P<0.001)。肿瘤浸润范围T 1~2期患者NLRP1 mRNA表达量高于T 3~4期[0.64(0.38,1.36)比0.40(0.06,0.65)](P<0.05);临床分期Ⅰ~Ⅱ期患者NLRP1 mRNA表达量高于Ⅲ~Ⅳ期患者[0.78(0.47,2.96)比0.40(0.97,0.82)](P<0.05)。RT-qPCR分析结果显示,CAL-27细胞中NLRP1的表达显著低于OMEC细胞(1.022±0.018比0.389±0.014)(t=47.702,P<0.001)。RT-qPCR结果显示,NLRP1过表达组NLRP1相对表达量高于阴性对照组(6.148±0.418比1.042±0.018)(t=21.137,P<0.001);Western blot结果显示,NLRP1过表达组NLRP1蛋白表达量高于阴性对照组(2.009±0.253比1.000±0.129)(t=6.155,P<0.001)。Transwell小室侵袭实验结果显示,pcDNA-NLRP1转染CAL-27细胞24 h后,NLRP1过表达组细胞侵袭数少于阴性对照组[(51.4±1.1)个比(77.0±5.0)个](t=8.693,P<0.001)。Transwell小室迁移实验结果显示,NLRP1过表达组细胞迁移数显著少于阴性对照组[(61.6±5.2)个比(73.1±3.3)个
Objective To detect the expression of nucleotide-binding oligomerization domain-like receptor protein 1(NLRP1)in oral squamous cell carcinoma(OSCC)tissues and its effect on the proliferation,migration and invasion of OSCC cell line CAL-27.Methods Primary OSCC tumor tissue samples and adjacent normal tissue samples were collected from 40 patients admitted to the Department of Oral and Maxillofacial Surgery,Guizhou Provincial People′s Hospital from Apr.2021 to Aug.2022.NLRP1 messenger RNA(mRNA)expression was detected in the 40 oral squamous carcinoma tissues and corresponding adjacent normal tissues by reverse transcription quantitative polymerase chain reaction(RT-qPCR),and the correlation analysis was conducted combined with patient clinicopathological parameters.The expression of NLRP1 in OSCC cell line CAL-27 and normal oral mucosal epithelial cells(OMEC)was detected by RT-qPCR.In vitro NLRP1 overexpression cell model was constructed by transient transfection of pcDNA-NPRP1 into OSCC CAL-27,and transfection of pcDNA-NC was used as a negative control group;RT-qPCR and Western blot were used to determine the efficiency of cell transfection.The effect of NLRP1 overexpression on cell proliferation of CAL-27 cells was determined by the cell counting kit-8 assay,while the effects of migration and invasion capabilities of CAL-27 were detected by scratch healing assay and Transwell assay.Results The NLRP1 mRNA expression in OSCC tissues was significantly lower than that in the paracanceral tissues[0.49(0.13,0.99)vs 1.02(1.00,1.03)](Z=4.224,P<0.001).The NLRP1 mRNA expression in the patients with tumor invasion stage T 1-2 was higher than that in the patients with tumor invasion stage T 3-4[0.64(0.38,1.36)vs 0.40(0.06,0.65)](P<0.05).The NLRP1 mRNA expression level in the stageⅠtoⅡpatients was higher than that in the stageⅢtoⅣpatients[0.78(0.47,2.96)vs 0.40(0.97,0.82)](P<0.05).RT-qPCR analysis showed that the expression of NLRP1 in the CAL-27 cells was significantly lower than that in the OMEC cells(1.022±0.018
作者
王芳
段晓峰
WANG Fang;DUAN Xiaofeng(Guizhou Medical University,Guiyang 550004,China;Department of Oral and Maxillofacial Surgery,Guizhou Provincial People′s Hospital,Guiyang 550499,China)
出处
《医学综述》
CAS
2023年第18期3731-3738,共8页
Medical Recapitulate
基金
贵阳市科技计划项目(筑科合同〔2018〕1-84号)。
关键词
口腔鳞状细胞癌
核苷酸结合寡聚化结构域样受体蛋白1
增殖
迁移
侵袭
Oral squamous cell carcinoma
Nucleotide-binding oligomerization domain-like receptor protein 1
Proliferation
Migration
Invasion
