Expression of exon 13 from the Ki-67 gene in human cells and tissues by digoxigenin-labelled mRNA in situ hybridization
10.3760/j.issn:0366-6999.2001.01.010
- VernacularTitle:人类细胞和组织Ki-67基因exon 13的转录和表达
- Author:
Yulian WU
1
;
Chenghong PENG
;
Hongwei SHEN
Author Information
1. 浙江大学附属第二医院
- Keywords:
in situ hybridization;
Ki-67;
mRNA;
MIB-1;
cell proliferation
- From:
Chinese Medical Journal
2001;114(1):48-53
- CountryChina
- Language:Chinese
-
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.
