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研究Ki 67基因在人类正常和异常细胞的mRNA转录和翻译 ,了解其在肿瘤细胞增殖中的作用。方法 以PCR扩增Ki 67基因的exon 13、codon 2的cDNA片断 ( 4 35bp) ,地高辛标计转录合成mRNA探针 ,原位杂交结合MIB 1免疫组化分析Ki 67基因的第 13外显子 (exon 13)在Hela细胞、结肠癌细胞、扁桃体和胰腺癌石蜡切片中的转录及翻译。结果 应用地高辛标记的mRNA原位杂交技术可成功地在Hela细胞、结肠癌细胞、扁桃体和胰腺癌的石腊切片检测到Ki 67mRNA信号 ,结肠癌和胰腺癌细胞均存在Ki 67基因的exon 13mRNA的过度转录 ,并且exon 13的转录与MIB 1所测Ki 67蛋白相一致。结论 本研究首次在人类组织和细胞中 ,应用原位杂交和免疫组化研究Ki 67基因外旋子 13的转录和翻译 ,研究发现结肠癌和胰腺癌Ki 67基因的exon 13的过度转录 ,并与其蛋白水平高度相关 ,提示该此基因表达异常 ,尤其是exon 13,在恶性肿瘤的发生和发展可能起一定作用。
To study the mRNA transcription and translation of Ki 67 gene in human normal and abnormal cells to understand its role in tumor cell proliferation. Methods The cDNA fragment of exon 13 and codon 2 of Ki 67 gene was amplified by PCR, and mRNA probe was synthesized by digoxigenin labeling. The in situ hybridization and MIB 1 immunohistochemistry were used to analyze the 13 th of Ki 67 gene Transcription and Translation of Exon 13 in HeLa Cells, Colon Cancer Cells, Amygdala and Pancreatic Cancer Paraffin Slices. Results Ki67 mRNA was detected successfully by digoxin labeled mRNA in situ hybridization in paraffin sections of Hela cells, colon cancer cells, tonsils and pancreatic cancer, while exon of Ki 67 gene existed in both colon cancer and pancreatic cancer 13 mRNA over-transcription, and the transcription of exon 13 is consistent with that of MIB 1 as measured by Ki 67 protein. Conclusions This study is the first to study the transcription and translation of Ki 67 gene exon 13 by in situ hybridization and immunohistochemistry in human tissues and cells. It was found that overexpression of exon 13 of Ki 67 gene in colon and pancreatic carcinomas was correlated with The protein level is highly correlated, suggesting that the gene expression abnormalities, especially exon 13, may play a role in the occurrence and development of malignant tumors.