AU2002343900A1 - Diagnostic method for cancer characterized in the detection of the deletion of G-CSF exon 3 - Google Patents

Description

DIAGNOSΗC METHOD FOR CANCER CHARACTERIZED IN THE DETECTION OF

THE DELETION OF G-CSF EXON 3

TECHNICAL FIELD

The present invention relates to a method of diagnosing cancer based on modified

features in granulocyte colony stimulating factor (G-CSF) mRNA or protein. More particularly,

the present invention relates to a diagnostic and prognostic method for cancer based on skipping of

the exon 3 region of the G-CSF gene at mRNA or protein levels, wherein skipping of G-CSF exon

3 is used as a diagnostic cancer marker.

PRIOR ART

Cancer is a leading cause of death in developed nations. For this reason, a major interest

in cancer therapy is to develop methods for early diagnosis and treatment of cancer. Typically, late-

stage cancer is almost incurable, whereas, at the early stage, cancer can be more effectively treated

and therapeutic methods for early-stage cancer are simpler. Therefore, there is an urgent need for

development of methods for accurately and quickly diagnosing cancer.

At present, cancer diagnosis is generally achieved by morphological analysis using

microscopes such as an optical microscope or electron microscope, irnmunohistochemical assays which detect proteins specifically expressed in caner tissues (Iran. Biomed. J. 3 (3 & 4): 99-101,

1999; and Lancet 2:483-6, 1986), or molecular analysis of abnormal biomolecules found in cancer

tissues, such as mutated genes. In comparison with the molecular diagnosis, the moφhological and

immunohistochemical diagnosis requires much longer time and higher cost. Because of comprising

a relatively simple procedure and yielding results in a short time, the molecular diagnosis methods are

a focus for developing novel diagnostic methods for cancer. Recently, a protein chip system for

diagnosing various cancers has been developed by Health Digit Inc. in Shanghai, China, and gained

approval for clinical tests from the Chinese State Drug Admistration (CSDA). Such an approval is

the first in the world (www.health-digit.com). However, the protein chip system does not use only a

biomarker to diagnose all kinds of cancer, but uses 10 or more proteins.

To effectively apply such diagnostic methods to cancer diagnosis, it is most important to

select cancer diagnostic markers capable of more accurately and easily discovering incidence of

cancer. As diagnostic cancer markers, several genes (Steve M. et al., J. Clin. Oncology 20:3165-

3175, 2002; and Sridlhar R. et al, J. Clin. Oncology 20:1932-1941, 2002) and proteins (Goessl et al.,

Urology 58:335-338, 2001; Zhou et al., Breast Cancer Res Treat 66:217-224, 2001; and CK Kim et

al., Korea Pat. Publication No. 2001-0061173) have been reported, and some of them are being

clinically used for diagnosis of cancer. The conventional cancer biomarkers are unable to detect all

kinds of cancer, as follows. The known cancer biomarkers which have low organ specificity, such as CEA, BFP, TPA and IAP, also, have low sensitivity, thus generating false positive data. Also, the

biomarkers which have high organ specificity, exemplified by AFP, PIVKA II, Esterase L CA19-9,

CA50, Span-1 antigen, CA15-3 and BCA 225, are useful only for target organs. Therefore, for

accurate, economical and simple diagnosis of cancer, there is an urgent need for development of new

markers capable of diagnosing a variety of cancers.

DISCLOSURE OF THE INVENTION

Leading to the present invention, the thorough and intensive research into a cancer biomaker capable of diagnosing a variety of cancers, conducted by the present inventors, resulted in the finding that exon 3 skipping occurs during transcription of the G-CSF gene in cancer patients, and use of G-CSF mRNA f agment or protein as a diagnostic cancer marker can achieve diagnosis of a variety of cancer, wherein the diagnosis is performed simply and quickly, as well as being economical.

In an aspect of the present invention, there is provided a mutated G-CSF mRNA fragment

having a deletion of an exon 3 region for use as a diagnostic cancer marker.

In another aspect of the present invention, there is provided a mutated G-CSF protein having a deletion of an amino acid sequence corresponding to an exon 3 region for use as a cancer diagnostic marker.

In a further aspect of the present invention, there is provided a microarray or membrane for diagnosis of cancer comprising (a) a DNA fragment corresponding to exon 3 of a G-CSF gene, and (b) at least one of DNA fragments corresponding to exons 1, 2, 4 and 5 of said G-CSF gene.

In a still further aspect of the present invention, there is provided a diagnostic agent for invention are given in Table 1, below.

TABLE 1

Normal and tumor cell lines used in the present invention

20