Cell proliferation inhibitory proteins and polynucleotides, antisense polynucleotides to the polynucleotides, cell proliferation inhibitors using the foregoing, cancer diagnostic agents, cancer therapeutic agents and compositions for gene therapy

a cell proliferation inhibitor and polynucleotide technology, applied in the field of gene information, can solve the problems of human cell aging, immortalization, or cancerization of human cells, and the correlation between these genes and aging,

Inactive Publication Date: 2006-12-07
HIROMI KUMON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009] In view of the aforementioned circumstances this invention aims at identifying the complete lengths cDNA of an immortalization...

Problems solved by technology

However, the correlation between these genes and the aging, imm...

Method used

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  • Cell proliferation inhibitory proteins and polynucleotides, antisense polynucleotides to the polynucleotides, cell proliferation inhibitors using the foregoing, cancer diagnostic agents, cancer therapeutic agents and compositions for gene therapy
  • Cell proliferation inhibitory proteins and polynucleotides, antisense polynucleotides to the polynucleotides, cell proliferation inhibitors using the foregoing, cancer diagnostic agents, cancer therapeutic agents and compositions for gene therapy
  • Cell proliferation inhibitory proteins and polynucleotides, antisense polynucleotides to the polynucleotides, cell proliferation inhibitors using the foregoing, cancer diagnostic agents, cancer therapeutic agents and compositions for gene therapy

Examples

Experimental program
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example 1

Cloning Immortalization-Related Genes

1. Culturing the First Generation Fibroblast Line

[0093] Human fibroblasts were prepared from a female embryo of 9 weeks old to form the first generation fibroblast line (KMS-6) (Namba, M. et al., Int. J. Cancer, 35: 275-280, 1985). The cells were cultured with an Eagle minimum medium (MEM available from GIBCO) containing 10% fetal bovine serum (FBS available from Sanko Pure Chemicals Co. Ltd.) or with a Dalbeco modified Eagle medium (DMEM available from GIBCO) under the conditions of 5% CO2 concentration and 37° C. After the cells were grown until almost confluent state in a plate for 5 to 7 days, cells were diluted ¼-fold and subcultured.

2. Establishing an Immortal Cell Line by Treatment with 60Co Radiation

[0094] KMS-6 cells that had been cultured semiconfluent were radiated with 60Co γ-ray having a dose of from 200 to 400 rads 13 times and were subjected to radiation treatment with a total of 2,800 rads. Subsequently, subculturing was rep...

example 2

REIC Clone

1. Screening the E. coli Library

[0096] To remove pseudo-positive clones from the E. coli library, 32P labeled cDNA probe was prepared based on mRNA of OUMS-24F, a different immortal cell line (Bai, L. et al., Int. J. Cancer, 53: 451-456, 1993); the probe was used to carry out colony hybridization. The hybridization resulted in the removal of clones that had proved positive and the production of about 30 immortal candidate clones.

2. DNA Sequencing

[0097] The DNA sequences of the clones obtained by screening were determined according to the Sanger method (Sanger F. et al., Proc. Natl. Acad. Sci. USA 74: 5463-5467, 1977). Consequently, clones were identified that had the aging-related gene sequences, including fibronectin, extracellular matrix proteins (e.g., α2 type I procollagen), collagenase, enzyme proteins (e.g., WS9-14), cell cycle regulating proteins (e.g., p21). Among these, it was found that REIC clone (D93) displaying no homology to known genes and being regard...

example 3

REIC Clone (10-1)

[0098] The DNA sequence of REIC clone (D93) predicted that the clone was not of the complete length cDNA. The polynucleotide having the DNA sequence set forth in SEQ ID NO:5 was used as a probe to screen a human heart cDNA library (BRL Inc.) for clones having cDNA fragments with longer DNA sequences. Consequently, there was obtained an REIC clone (10-1) that was predicted to carry the full-length cDNA containing the 5′-region of REIC (D93). DNA sequencing was conducted on this cDNA clone and the entire DNA sequence was determined. The DNA sequence of this clone is shown in SEQ ID NO:3 in the Sequence Listing.

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Abstract

The genes the expression of which is reduced or disappeared in immortal cells including cancer cells are isolated, their DNA sequences are determined, the genes are expressed to produce cell proliferation inhibitory proteins, and the genes and the proteins are utilized as agents for diagnosis or treatment, including the genetic diagnosis of or the gene therapy of diseases such as cancer.

Description

RELATED APPLICATIONS [0001] This Application is a Rule 1.53(b) Divisional Patent Application of U.S. patent application Ser. No. 10 / 130,360 filed on Oct. 7, 2002 and having a 35 U.S.C. §371(c) date of Oct. 7, 2002, which is a National Phase of International Application No. PCT / JP00 / 15879 filed Aug. 30, 2000 that designated the U.S. and was not published under PCT Article 21(2) in English, and this divisional application claims, via the aforesaid U.S. Application and the aforesaid International Application, the foreign priority benefit of and claims the priority from Japanese Application No. 1999-330604, filed Nov. 19, 1999, the complete disclosures of all the prior applications, including any and all sequence listings (as well as paper copy, disc copy and / or diskette copy), are incorporated herein by reference.TECHNICAL FIELD [0002] This invention relates to genetic information that can be useful in the treatment and diagnosis of diseases caused by the immortalization of cells such ...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/861C12Q1/68A61K31/713A61K35/76A61K35/761A61K38/00A61P35/00A61P43/00C07K14/47C12N15/09C12N15/113C12N15/12G01N33/574
CPCA61K38/00C07K14/4703A61K48/00A61P1/16A61P9/10A61P13/12A61P17/00A61P31/12A61P31/16A61P31/20A61P35/00A61P35/02A61P35/04A61P43/00A61K35/76A61K35/761A61K48/0016A61K48/0058A61K48/0075
Inventor NAMBA, MASAYOSHITSUJI, TOSHIYA
Owner HIROMI KUMON
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