膀胱癌尿液脱落细胞角质素mRNA表达检测方法的建立  

Established a method of detecting the expression of CK8 and CK20 mRNA in exfoliated bladder cells

作　　者：燕翔[1] 丁强[1] 张元芳[1] 蓝斐[2] 

机构地区：[1]复旦大学泌尿外科研究所 [2]上海市肿瘤研究所,上海200032

出　　处：《临床肿瘤学杂志》2003年第3期161-163,共3页Chinese Clinical Oncology

摘　　要：目的 :建立采用RT PCR对膀胱癌尿液脱落细胞CK8和CK2 0的mRNA表达进行分析的实验方法。方法 :从 2 2例膀胱癌患者尿液脱落细胞中提取总RNA ,采用RT PCR和特异性引物进行逆转录、扩增 ,并用琼脂糖凝胶电泳进行分析。结果 :从所有标本中提取的总RNA均有较好纯度和均一性 ;2 2例标本均表达CK8mRNA ,1 8例标本表达CK2 0mRNA。结论 :RT PCR是一种敏感、快捷的检测膀胱癌尿液脱落细胞中CK8、CK2 0mRNA表达的方法 ,可采用此方法从分子水平对膀胱癌患者尿液脱落细胞表达产物进行无创性检测 ,为研究膀胱癌的发病机制。Objective:To establish an experimental method for detecting the expression of CK8 (cytokeratin 8) and CK20 (cytokeratin 20) mRNA in exfoliated bladder cells with reverse transcription polymerase chain reaction (RT PCR).Methods:Cells were obtained from urine of 22 patients with bladder cancer confirmed by bladder biopsy. TRIzol reagent was used to isolate total RNA from the exfoliated bladder cells. The Access RT PCR system was performed to reverse transcript the mRNA and amplify the cDNA with special primers of CK8 and CK20 respectively. RT PCR products were analyzed by agarose gel electrophoresis and ethidium bromide staining against PCR size markers. The results were documented with a camera.Results:The total RNA isolated from the exfoliated bladder cells was integral and pure and the yield of total RNA was also ideal. CK8 amplification (244 base pairs) and CK20 amplification (370 base pairs) were obtained with total RNA. All the 22 specimens express the mRNA of CK8 and in which 18 specimens express the mRNA of CK20.Conclusion:RT PCR is a sensitive, rapid and noninvasive method of detecting the expression of CK8 and CK20 mRNA in exfoliated bladder cells. It may provide an experimental method for studying the mechanism, biological behavior and early diagnosis of baldder cancer.

关 键 词：膀胱癌 尿液脱落细胞 MRNA表达 检测 细胞角质素 实验 

分 类 号：R737.14[医药卫生—肿瘤]