荧光定量RT-PCR检测Rac1 mRNA基因及其与胃癌转移的关系  

The relationship between Rac1 mRNA gene levels detected by FQ-RT-PCR and the metastasis in gastric carcinomas

作　　者：高丹丹[1] 续薇[1] 刘力华[2] 张明威[3] 罗微 

机构地区：[1]吉林大学第一医院检验科,长春130021 [2]吉林大学第一医院中心研究室,长春130021 [3]吉林大学第二医院普外科 [4]吉林市铁路医院

出　　处：《中华检验医学杂志》2010年第6期502-506,共5页Chinese Journal of Laboratory Medicine

基　　金：吉林省科技厅自然科学基金资助项目（200705123）

摘　　要：目的 探讨肿瘤转移相关基因Rac1水平与胃癌转移的相关性,并评价分析Rac1 mRNA基因在胃癌转移和区分胃良、恶性病变中的价值.方法 用荧光定量RT-PCR TaqMan探针技术,选择Rac1基因目的 片段,与pET-20b（＋）载体连接构建重组质粒,建立Rac1 mRNA荧光定量RTPCR标准曲线,并用于检测52份胃癌、癌旁组织及12份胃良性病变组织标本中Rac1 mRNA的含量,分析评价其与胃癌转移的关系.结果 胃癌组织中Rac1 mRNA表达阳性率和含量为100.0%,7.41×103 （3.50×105 ～4.36×106）拷贝/μl,癌旁组织为46.2%,0（0～1.73×104）拷贝/μl,良性病变组织为33.3%,0（0～3.55×103）拷贝/μl,3组间阳性率和Rac1 mRNA含量水平差异均有统计学意义（x2=43.16,x2k-w=64.19,P均＜ 0.01）.当ROC曲线确定荧光定量RT-PCR检测Rac1含量的最佳临界值为1.73×104 拷贝/μl时,诊断胃癌组织标本的敏感度为88.5%,特异度为100.0%;当最佳临界值为7.49×103 拷贝/μl时,诊断胃癌是否发生淋巴结转移的敏感度为57.9%,特异度为78.6%.结论 荧光定量RT-PCR检测Rac1 mRNA对鉴别胃良、恶性病变及评价胃癌转移均有参考价值.Objective To investigate the relationship between tumor metastasis-related Rac1 mRNA expression levels and gastric carcinomas metastasis, and to investigate the significance of Rac1 as a tumor marker for the evaluation of gastric carcinomas metastasis and distinguish between benign and malignant lesions. Methods This experiment used fluorescence quantitative RT-PCR TaqMan probe technology, chose Rac1 target gene fragment, which was cloned into the pET-20b （＋） vector, constructed recombinant plasmid, and established the Rac1 mRNA fluorescence quantitative RT-PCR standard curve. And then the Rac1 mRNA levels were detected in 52 cases of gastric carcinoma tissues,52 cases of para-carcinoma tissues and 12 cases of benign gastric disease tissues. Its association with the metastasis of gastric carcinomas was analyzed. Results The positive rates and levels （median, P25-P75） of Rac1 mRNA were 100. 0% ,7.41 ×105 （3. 50 × 105-4. 36 × 106） copies/μl in gastric carcinomas, 46. 2% ,0（0-1.73 × 104） copies/μl in parscarcinoma tissues, 33. 3%, 0（0-3.55 × 103） copies/μl in benign tissues. The positive rates and leves（x2 =43.16,x2Xk-w = 64. 19, P ＜0. 01）among the three groups were significantly different. When the critical value of Rac1 mRNA level was 1.73 × 104 copies/μl determined by ROC, the sensitivity and specificity were 88. 5% and 100% respectively. When the critical value of Rac1 level was 7.49 × 105 copies/μl, the sensitivity and specificity were 57.9% and 78. 6% respectively. ConclusionThe Rac1 mRNA level detected with fluorescence quantitative RT-PCR has reference value to distinguish between benign and malignant lesions and evaluate gastric carcinoma metastasis.

关 键 词：胃肿瘤 肿瘤转移 Ras相关的C3肉毒素底物1 逆转录聚合酶链反应 

分 类 号：R735.2[医药卫生—肿瘤]