荧光定量PCR技术检测喉鳞状细胞癌标本中Skp2基因表达

Detection of Skp2 mRNA gene expression using fluorogenic probe quantitative RT-PCR method in laryngeal squamous cell carcinoma

目的:探讨细胞S相激酶相关蛋白(Skp2)基因在喉鳞状细胞癌发生、发展中的作用。方法:应用荧光定量逆转录PCR(FQ-PCR)技术检测40例喉鳞状细胞癌及10例癌旁正常组织中Skp2 mRNA拷贝数,并分析其与临床相关因素的关系。结果:40例喉鳞状细胞癌Skp2 mRNA中位拷贝数为6 622.54 copy/μg RNA,10例癌旁正常组织为0 copy/μg RNA,两组差异有统计学意义(P<0.01);喉鳞状细胞癌和癌旁正常组织Skp2 mRNA阳性表达率分别为50%和0,差异有统计学意义(P<0.01)。喉鳞状细胞癌中颈淋巴结转移组Skp2 mRNA中位拷贝数为617 138.4 copy/μg RNA,颈淋巴结无转移组为0 copy/μg RNA,两组差异有统计学意义(P<0.05);喉鳞状细胞癌中颈淋巴结转移组和颈淋巴结无转移组Skp2 mRNA阳性表达率分别为100.00%和35.48%,差异有统计学意义(P<0.01)。结论:Skp2基因可能与喉鳞状细胞癌颈淋巴结转移有关,FQ-PCR为一精确检测喉鳞状细胞癌Skp2 mRNA表达的方法,Skp2 mRNA表达水平可能成为预测喉鳞状细胞癌颈淋巴结转移的更加灵敏的指标。

Objective： The current study was designed to examine the expression of Skp2 gene in laryngeal squamous cell carcinoma （LSCC） and to investigate the role of Skp2 gene in tumorigenesis and progression of LSCC. Method..FQPCR method was used to examined the expression of Skp2 gene in 40 LSCC and 10 normal laryngeal mucosa tissues, and relationship between its expression and clinical biological factors of patients with LSCC was analyzed. Result：The median copy number of Skp2 mRNA expression in LSCC was 6 622.54 copy/ ug RNA, the median copy number of Skp2 mRNA expression in normal laryngeal mucosa tissues was 0 copy/ug RNA, there was a very significant difference between them （P〈0.01） ; The positive rate of Skp2 mRNA expression in LSCC and adjacent normal laryngeal tissue were 50 %0, 0, respectively （P〈0.01）. The median copy number of Skp2 RNA expression in LSCC with cervical lymph node metastasis was 617 138.4 copy/ug RNA, the median copy number of Skp2 mRNA expression in LSCC without cervical lymph node metastasis was 0 copy/ug RNA, there was a very significant difference between them （P〈0.05） ; The positive rate of Skp2 mRNA expression in LSCC with and without cervical lymph node metastasis were 100.00%, 35.48%, respectively （P〈0.01）. Conclusion： Skp2 gene might have relation with the cervical lymph node metastasis of LSCC. FQ-PCR is an accurate assay to detecting expression of Skp2 mRNA in patient with LSCC. The level of Skp2 mRNA expression might be a new and more accurate marker, and it can be used to predict cervical lymph node metastasis of LSCC.