EGFR与CK20 mRNA在卵巢上皮性肿瘤中表达及意义.docVIP

EGFR与CK20 mRNA在卵巢上皮性肿瘤中表达及意义 　　【摘要】 目的研究表皮生长因子受体 (epidermal growth factor receptor, EGFR) 和细胞角质蛋白20(cytokeratin-20,CK20)在卵巢上皮性肿瘤中的表达及相关性,探讨二者在卵巢肿瘤发生、发展和转移中的作用。方法 应用逆转录聚合酶链式反应(RT-PCR)检测12例正常卵巢(对照组)、18例卵巢良性肿瘤(良性组)、33例卵巢癌(恶性组)组织中EGFR 、CK20 mRNA的表达,分析其与临床病理特征的关系。结果 卵巢癌组织中EGFR mRNA和CK20 mRNA的阳性表达率显著高于正常卵巢及卵巢良性肿瘤组织(P0.05); CK20 mRNA的表达与手术病理分期、淋巴结转移有关(P0.05);)EGFR与CK20表达之间有显著相关性(P 　　【关键词】 卵巢癌;表皮生长因子受体;细胞角质蛋白 20;逆转录聚合酶链式反应 　　 　　Expression and Significance of EGFR and CK-20 in Ovarian Carcinoma 　　 　　ZOU Zhong-xiang,XU Jie,SHA Xiu-min,et al. 　　Department of Obstetrics and Gynecology,Yantai City Laiyang Central Hospital,Laiyang 265200, China 　　 　　【Abstract】 Objective To investigate the expression and significance of epidermal growth factor receptor (EGFR) and cytokeratin-20 (CK20) in ovarian epithelial tumor, and to explore their roles in the tumorigenesis, development and metastasis of ovarian tumor. Methods The expression of EGFR and CK20 were detected by reverse transcription polymerase chain reaction (RT-PCR) in 12 cases of normal ovary tissue, 18 cases of benign ovarian tumor and 33 cases of primary ovarian carcinoma. Analyzed the relationship between the two genes and the different clinicopathological features. Results The positive rates of EGFR mRNA mRNA and CK20 mRNA were significantly higher in m alignant ovarian specimens than that in the normal and benign ones (P0.05). There was distinct correlation between EGFR mRNA and CK20 mRNA expression (P 本文为全文原貌 未安装PDF浏览器用户请先下载安装 原版全文 　　1.2.3 总RNA提取 按RNA抽提操作手册(大连宝生物工程公司)的方法提取组织总 RNA。提取的RNA 经琼脂糖凝胶电泳鉴定其完整性,通过紫外分光光度计测定260 nm与280 nm吸光度值而计算RNA的纯度与浓度。 　　1.2.4 逆转录反应,cDNA的合成 反应体系:dNTP Mixture 1ul,Template RNA 1 μg, RNase Free dH2O至10ul,65℃孵育5 min后置于冰上,加入5×Prime Script Buffer 4 μl, RNase Inhibitor 0.5 μl, Prime Script RTase 0.5ul,RNase Free dH2O 5ul, 30℃10 min,42℃20 min,95℃5 min后置于冰上。逆转录出的cDNA用于PCR反应或于-20℃保存备用。 　　1.2. 5 聚合酶链式反应 取上述逆转录反应液2 μl,加入10×PCR Buffer 2 μl,dNTP Mixture 0.8 μl,TaKaRaExTaqHS 0.2 μl,上下游引物各1