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.
出处
《中华外科杂志》
CAS
CSCD
北大核心
1998年第12期732-734,I150,共4页
Chinese Journal of Surgery
关键词
胰腺肿瘤
基因表达
KI-67基因
Pancreatic neoplasms Gene expression regulation In situ hybridization Immunohistochemistry
