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Human cancer suppressor gene, protein encoded therein

A technology of tumor suppressor protein and tumor suppressor gene, applied in the field of human tumor suppressor gene and its encoded protein

Inactive Publication Date: 2008-05-21
金弦起
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, p53 mutations have been reported to total only in the range of 30% of liver cancers in the United States and Western Europe, and there are no hotspots where such mutations occur frequently (Szymanska, K. & Hainaut, P. Acta Biochimica Polonica, 50 , 231-238(2003))

Method used

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  • Human cancer suppressor gene, protein encoded therein
  • Human cancer suppressor gene, protein encoded therein
  • Human cancer suppressor gene, protein encoded therein

Examples

Experimental program
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Effect test

Embodiment 1

[0247] Example 1 : Total RNA isolation and mRNA differential display

[0248] The differential expression patterns of observed genes in normal breast tissue, primary breast cancer tissue and breast cancer cell lines are as follows.

[0249] Normal breast tissue samples were obtained from breast cancer patients during mastectomy, and primary breast cancer tissue samples were obtained during radical mastectomy from breast cancer that had not undergone radiation therapy and / or anticancer chemotherapy prior to surgical treatment patients get. MCF-7 (American Type Culture Collection, ATCC No. HTB-22) was used as a human breast cancer cell line. Total RNA was isolated from these tissues and cells in the same manner as described in Reference Examples.

[0250] For the differential display of GIG15 mRNA, bone marrow tissue was also obtained from normal individuals and, at the time of bone marrow biopsy, from leukemic patients who had not previously received anticancer chemotherapy...

Embodiment 2

[0377] Example 2 : cDNA library screening

[0378] According to published texts (Feinberg, A.P. and Vogelstein, B., Anal.Biochem., 132 , 6-13 (1983)) method respectively the cDNA fragments FC33, FC42, FC59, GV2, H-AP8, HP71, HP115, HP3, FC48, FC82, FC86, FC35, FC38, FC122, FC126, HP79, HP85, FC102, CA161, HP29, HP95, HP96, CA324, FC101, FC22, L927, HP102, FC123 and FC47 marked to obtain 32 P-labeled probes FC33, FC42, FC59, GV2, H-AP8, HP71, HP115, HP3, FC48, FC82, FC86, FC35, FC38, FC122, FC126, HP79, HP85, FC102, CA161, HP29, HP95, HP96 , CA324, FC101, FC22, L927, HP102, FC123 and FC47cDNA, according to the method described in the open text (Sambrook, J. et al., Molecular Cloning: A Laboratory manual, NewYork: Cold Spring Harbor Laboratory (1989) 32 The P-labeled probe was subjected to plaque hybridization with the phage λgt11 human embryonic lung fibroblast cDNA library (Miki, T. et al., Gene, 83, 137-146 (1989)) to obtain the full length of the human tumor suppressor g...

Embodiment 3

[0466] Example 3 : Northern blot of the GIG gene

[0467] 3-1. GIG8, GIG10, GIG13, GIG30, GIG32, GIG33, GIG34, GIG35, GIG38, GIG39, GIG43, PIG49, PIG51, GIG44, and GIG31

[0468] To evaluate the expression levels of GIG and PIG genes, Northern blots were performed as follows.

[0469] 20 μg of each total RNA sample obtained from three normal breast tissues, three primary breast cancer tissues and the breast cancer cell line MCF-7 in Example 1 were denatured and electrophoresed in 1% formaldehyde agarose gel, The resulting agarose gel was then transferred to a nylon membrane (Boehringer-Mannheim, Germany). Then the nylon membrane was labeled with the partial sequences FC33, FC42, FC59, FC48, FC82, FC86, FC35, FC38, FC122, FC126, FC102, FC101, FC22, FC123 and FC47 of the full-length GIG cDNA using the Rediprime II random primer labeling system ( Amersham, UK) 32 P-labeled random primer probes were hybridized overnight at 42°C. The Northern blotting procedure was repeat...

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Abstract

Disclosed are a human cancer suppressor gene, a protein encoded therein, an expression vector containing the same and a microorganism transformed with the vector. The cancer suppressor gene of the present invention may be effectively used for diagnosing, preventing and treating human cancers.

Description

technical field [0001] The invention relates to a human tumor suppressor gene, its coded protein, an expression vector containing the gene and cells transformed with the vector. Background technique [0002] Tumor suppressor gene products play a role in inhibiting normal cells from transforming into certain cancer cells, so the loss of this function of tumor suppressor gene products makes normal cells become malignant transformations (Klein, G, FASEB J., 7, 821-825 (1993)). In order for a cancer cell to develop into cancer, the cell should lose the ability to control the normal copy number of a tumor suppressor gene. Variations in the coding sequence of the p53 tumor suppressor gene were found to be among the most prevalent genetic variations in human cancers (Bishop, J.M., Cell, 64 , 235-248 (1991); and Weinberg, R.A., Science, 254 , 1138-1146 (1991)). [0003] However, because p53 mutations reported in breast cancers amount to within 30% of all breast cancers, it is pr...

Claims

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Application Information

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IPC IPC(8): C07K14/47
CPCC07K14/4703A61P35/00A61P35/02A61P35/04A61P43/00A61B5/16
Inventor 金弦起金振宇
Owner 金弦起
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