摘要
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 RNA probe of KAIl gene for study of the expression of the gene in pancreatic cancer. Methods The RNA probe was prepared by transcription in vitro with the linearizational plasmid as templet and through binding DIG 11 UTP into the RNA produced. With the probe, KAIl mRNA expression was studied by chemiluminescent technique and Northern blot in 12 normal pancreas specimens and 30 pancreatic cancer specimens. Results The RNA probe was highly sensitive. Northern blot analysis using the probe showed that KAIl mRNA presented stable expressed signals (2. 4 kb) in 25 specimens of pancreatic cancer without metastasis, weak signals in 5 specimens with metastasis, and weak or negative signals in normal pancreas specimens. Conclusions The RNA probe of KAIl gene labeled by this technique was high sensitive. Low expression of KAIl gene was related with metastasis of pancreatic cancer.
出处
《胰腺病学》
2002年第3期159-161,共3页
Chinese JOurnal of Pancreatology
基金
国家自然科学基金(39970334)
