口腔颌面鳞状细胞癌相关基因的筛选与定量验证

目的 筛选和验证与口腔颌面鳞状细胞癌密切相关的基因，为口腔鳞癌寡核苷酸功能芯片的制作提供可靠的靶标基因。方法 选用近5年口腔鳞癌基因表达谱芯片检测方面的文献资料，按标准筛选可能与口腔鳞癌相关的基因，用22对临床配对肿瘤与正常黏膜组织，采用实时定量PCR技术验证候选基因的表达情况，结合 内参照3-磷酸甘油醛脱氢酶（GAPDH）管家基因进行相对定量分析。结果 在课题组前期筛选到38个口腔鳞癌密切相关基因的基础上，又筛选出8个相关基因，其中富含半胱氨酸酸性分泌蛋白（SPARC）、血小板源生长因子A（PDGF—A）、丝氨酸蛋白酶抑制剂E（SERPIN口）、转化生长因子Cβ1（TGF—β1）、血管内皮生长因子C（VEGF—C）基因分别在16个以上肿瘤组织标本中过表达，细胞角蛋白15（CK15）基因在19个肿瘤标本中低表达，且肿瘤与正常黏膜组织存在差异，具有统计学意义（P〈0．01）。细胞周期D1蛋白基因（CCND1）和细胞凋亡相关的抑制蛋白（BIRC3）基因在肿瘤组织中过表达，但与正常组织差异无统计学意义（P〉0．05）。结论 SPARC、PDGF—A、SERPIN口、TGF-β、、VEGF—C、CK15是与口腔鳞癌密切相关的基因，可作为口腔颌面鳞癌寡核苷酸功能芯片制作的靶标基因。

Selection and quantitative detection of target genes in oral squamous cell carcinoma

OBJECTIVE: To select and identify the target genes related to oral squamous cell carcinoma (OSCC) and provide target genes for designing oligo-nucleotide functional microarray of OSCC. METHODS: Genes possibly related to oral squamous cell carcinoma were selected from the 5 years' published data of differently expressed profiles with microarray testing in OSCC. Then mRNA expression of selected genes were evaluated by real time quantitative polymerase chain reaction (RT-PCR) in 22 cases of OSCC, including tumor tissues and paried normal mucosas and quantified according to an internal control GAPDH. RESULTS: Eight genes were tested. The overexpression of SPARC, PDGF-A, SERPINE1, TGF-beta(1) and VEGF-C genes were measured in 16, 18, 16, 20, 18 cases of tumor specimens, respectively. The expression of CK15 gene was lower than that of its normal tissue. There were overexpression of CCND1, BIRC3 in tumor tissues, but there was no significant difference of CCND1 and BIRC3 expression between tumor tissue and normal tissue (P > 0.05). CONCLUSIONS: SPARC, PDGF-A, SERPINE1, TGF-beta(1), VEGF-C and CK15 genes were closely related to tumor progress of OSCC. They can be used as the target genes for designing oligo-nucleotide functional microarray of OSCC.