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目的 制备DIG标记KAIl RNA探针和分析KAIl基因在胰腺癌中的表达。方法 以线性KAIl基因DNA质粒为模板,采用体外转录法掺入DIG-11-UTP制备RNA探针,并通过化学发光法检测。应用该探针通过Northern blot对12例正常胰腺组织和30例原发性胰腺癌组织中的KAIl基因mRNA进行分析。结果 该方法标记的KAIl RNA探针浓度为1 ug/μl时即可检测。Northern bolt分析发现25例无转移的胰腺癌中KAIl基因在2.4kb处存在明显的杂交信号,5例发生转移的晚期胰腺癌杂交信号较弱,正常胰腺组织中该基因的表达水平呈阴性或弱阳性。结论 本法标记的KAIl RNA探针灵敏度高,KAIl基因低表达与胰腺癌的转移有关。
Objective To prepare DIG-labeled KAI1 RNA probe and analyze the expression of KAI1 gene in pancreatic cancer. Methods Linear KAI1 DNA plasmid was used as a template to prepare RNA probe by in vitro transcription using DIG-11-UTP and detected by chemiluminescence. This probe was used to analyze KAI1 mRNA in 12 cases of normal pancreatic tissue and 30 cases of primary pancreatic cancer by Northern blot. Results The KAI1 RNA probe labeled with this method can be detected at a concentration of 1 μg / μl. Northern bolt analysis showed that 25 cases of non-metastatic pancreatic cancer KAIl gene at 2.4kb at the presence of a significant hybridization signal, 5 cases of metastatic advanced pancreatic cancer hybridization signal is weak, normal pancreatic tissue expression was negative or weak Positive. Conclusion The KAIl RNA probe labeled by this method is highly sensitive and the low expression of KAI1 gene is associated with the metastasis of pancreatic cancer.