中华医学杂志（英文版）2001，Vol.114Issue(1)：48-53,6.
人类细胞和组织Ki-67基因exon 13的转录和表达
Expression of exon 13 from the Ki-67 gene in human cells and tissues by digoxigenin-labelled mRNA in situ hybridization
摘要
Abstract
Objective To get insight on the regulatory mechanism of Ki-67 gene expression in malignant cell cycle. Methods Non-radioactive in situ hybridization (ISH) was undertaken, combined with immunohistochemistry to study the Ki-67 gene transcription and translation in various human cells and tissues. HeLa cells and fresh colon cancer cells, tonsil, normal pancreas and pancreatic cancer tissues were used in this study. A 435?bp cDNA fragment located in exon 13 of the Ki-67 antigen gene was amplified by polymerase chain reaction (PCR). Digoxigenin-labelled antisense and sense RNA probes were prepared for detecting Ki-67 mRNA, combined with MIB-1 immuno～histochemistry. Results Successful localization of Ki-67 mRNA in human HeLa cells, colon cancer cells, tissues specimen of the tonsil and pancreatic cancer tissue sections was accomplished by digoxigenin-labelling in situ hybridization technique. ISH to colon cancer cells and pancreatic cancer tissue slides showed that much stronger cytoplasm and perinuclear mRNA signals of the Ki-67 gene were present in malignant cells than in normal cells, which was in accordance with MIB-1 nuclear protein signals. Conclusions A sensitive and practical in situ hybridization method for the analysis of Ki-67 antigen mRNA in human cell and tissue was developed. Abnormal transcription of exon 13 of Ki-67 gene might be responsible for malignant cell proliferation in colon and pancreatic cancer.关键词
Ki-67基因 mRNA原位杂交 MIB-1 细胞增殖Key words
in situ hybridization/Ki-67/mRNA/MIB-1/cell proliferation分类
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吴育连,彭承宏,沈宏伟..人类细胞和组织Ki-67基因exon 13的转录和表达[J].中华医学杂志（英文版）,2001,114(1):48-53,6.基金项目
This project was supported by both the National Natural Science Foundation of China (No. 39600141) and the Natural Science Foundation of Zhejiang Province (No. 396498). Remains of great interest in cellular molecular biology. （No. 39600141）
