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机构地区:[1]浙江医科大学附属第二医院外科实验室
出 处:《中华外科杂志》1998年第12期732-734,I150,共4页Chinese Journal of Surgery
摘 要:目的应用原位杂交结合免疫组化分析胰腺癌Ki67基因的mRNA转录和蛋白翻译,研究该基因结构和功能表达的关系。方法胰腺癌40例,正常胰腺及良性病变9例;扁桃体组织及Hela细胞作为阳性对照。通过地高辛标记的Ki67cRNA探针原位杂交和Ki67等效单克隆抗体(MIB1)的免疫组化分析该基因的mRNA转录和蛋白翻译。结果正常胰腺及良性病变Ki67指数均小于20%;胰腺高分化腺癌、低分化腺癌各20例,Ki67指数大于20%的分别为5例和12例。统计分析胰腺良恶性病变及不同分化类型间的Ki67指数,其差异有显著性意义(P<001)。结论人类胰腺癌存在Ki67基因的过度表达,表达强度与分级密切相关。Objective To investigate, at transcriptional and translational level in situ, the abnormal gene expression of the Ki 67 protein in pancreatic carcinoma specimens. Method 40 pancreatic cancer specimens, 5 normal pancreatic and 4 chronic pancreatitis tissues were studied.A 435 bp cDNA fragment located in codon 2, exon 13 of the Ki 67 antigen gene was amplified by the polymerase chain reaction (PCR). The DIG labeled cRNA probes were transcribed using a commercially available DIG RNA labeling kit. Localization of the Ki 67 protein and the specific mRNA were studied by combining immunohistochemistry (ICH) with (DIG) labeled in situ hybridization (ISH). Result The Ki 67 protein mRNA in formalin fixed and paraffin embeded pancreatic tissue sections was successfully localized.Analyzing the Ki 67 mRNA transcription in 17 pancreatic cancer specimens with Ki 67 ICH labeling index >20%, we found that stronger mRNA signals were also observed in poorly differentiated specimens with Ki 67 index >50% than in those well differented with ICH labeling index 20%~50%. A high expression of both mRNA and protein was observed in poorly differentiated pancreatic adenocarcinomas. Conclusion Abnormal overexpression of the gene encoding Ki 67 protein was detected not only at the protein level, but also at the mRNA level in pancreatic tumors. The abnormal overexpression of the Ki 67 protein might be correlated with the central part,exon 13, of the gene.
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