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Composition and method for diagnosing kidney cancer and estimating kidney cancer patient's prognosis

A composition and kidney cancer technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problem of low specificity and achieve the effect of improving prognosis

Inactive Publication Date: 2008-11-05
TORAY IND INC +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, as proteins known to increase in human kidney cancer, there are MMP2 (Non-Patent Document 3), which increases the motility of cancer cells by decomposing the extracellular matrix, and TNFRSF7 (Non-Patent Document 3), which is known to increase in renal dysfunction. Document 4), in addition, there are PDE8B (Patent Document 2), FLOT1 (Patent Document 3), CD5 (Non-Patent Document 5), ECM1 (Patent Document 4), etc., however, relatively speaking, its specificity is not Not high, the highly sensitive kidney cancer detection method obtained only by its expression level has not been used clinically

Method used

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  • Composition and method for diagnosing kidney cancer and estimating kidney cancer patient's prognosis
  • Composition and method for diagnosing kidney cancer and estimating kidney cancer patient's prognosis
  • Composition and method for diagnosing kidney cancer and estimating kidney cancer patient's prognosis

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preparation example Construction

[0313]

[0314] (1) Immunization and collection of antibody-producing cells

[0315] An immunogen composed of the target polypeptide is administered to mammals such as rats, mice (eg, inbred mouse Balb / c), rabbits, and the like. The dose of the immunogen administered at one time can be appropriately determined according to the species of animal to be immunized, the route of administration, etc., and it is about 50 to 200 μg per animal. Immunization is mainly performed by injecting immunogens subcutaneously or intraperitoneally. In addition, the interval of immunization is not particularly limited. After the primary immunization, booster immunizations are performed 2 to 10 times, preferably 3 to 4 times, at intervals of several days to several weeks, preferably at intervals of 1 to 4 weeks. After the initial immunization, use ELISA (Enzyme-Linked Immuno Sorbent Assay (Enzyme-Linked Immuno Sorbent Assay)) method, etc. to repeatedly measure the antibody value in the serum of t...

Embodiment 1

[0479] (1) Clinicopathological observation of the experimenter

[0480] Tissues were obtained from 31 Japanese kidney cancer patients undergoing nephrectomy with informed consent. Visual and / or histopathological observation was performed on the resected tissue slices, and the kidney cancer tissue was immediately frozen and stored in liquid nitrogen.

[0481] (2) Extraction of total RNA and preparation of cDNA

[0482] As a sample, the tissue of the kidney cancer lesion part among the kidney tissues of the kidney cancer patient was used. Total RNA was prepared from each tissue using Trizol reagent (Invitrogen) according to the manufacturer's recommended manual.

[0483] And use oligo(dT) primers and random nonamers, use the CyScribe First-StrandcDNA Labeling kit (GE Healthcare, Japan), according to the manufacturer's recommended operation manual, carry out reverse transcription reaction on 1 μg of total RNA obtained by the above method . Cy3-dUTP (GE Healthcare) was added t...

Embodiment 2

[0500] (1) Identification of proteins in the plasma of healthy people and kidney cancer patients

[0501] Before nephrectomy and 1 month after nephrectomy, EDTA-supplemented plasma fractions were obtained from five Japanese kidney cancer patients aged 50-70 years.

[0502] Plasma was filtered through a filter with a pore size of 0.22 μm to remove impurities and adjusted so that the protein concentration was 50 mg / mL. The plasma was further injected into 25 mM ammonium bicarbonate solution (pH 8.0) for dilution to a concentration of 12.5 mg / mL, and molecular weight cut-off was performed with a hollow fiber filter (Toray, Japan). Use ProteomeLab (registered trademark) PF2D System (Beckman Coulter Company) reversed-phase chromatography to separate the cut-off plasma sample (1.8mL total, containing up to 250μg protein) into 7 fractions, freeze-dried, and then dissolved in 100μL 25mM bicarbonate Ammonium solution (pH 8.0). This sample was digested with trypsin in an amount of 1 / 5...

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Abstract

A composition for detecting and diagnosing kidney cancer, diagnosing the metastasis of kidney cancer and / or estimating the prognosis of kidney cancer containing one or more polynucleotides selected from the group consisting of a polynucleotide, which shows a variation in the expression level in a kidney cancer cell originating in a patient with poor prognosis compared with a kidney cancer cell of a patient having favorable prognosis, its variant or a fragment thereof, or an antibody, which binds specifically to such a polypeptide showing a variation in the expression level as described above or its fragment, or a fragment thereof; a kit; a DNA chip; and utilization of the same.

Description

technical field [0001] The present invention relates to a composition for determination (diagnosis) that is useful for predicting the prognosis of a subject and / or for predicting or detecting renal cancer metastasis; a method for predicting or determining the prognosis of a subject using the composition, and / or using A method for predicting or detecting renal cancer metastasis by the composition; and a kit for predicting the prognosis of a subject and / or predicting renal cancer metastasis by using the composition. Background technique [0002] The kidney is an important urinary organ, which has the function of filtering blood to produce urine, thereby discharging the waste in the living body to the outside of the body. In addition, it is also an important endocrine organ, which can produce hormones such as angiotensin, which controls blood pressure, and erythropoietin, which is a hematopoietic factor for red blood cells. [0003] Tumors that occur in the kidney include rena...

Claims

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Application Information

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IPC IPC(8): C12N15/00C12N15/09C12Q1/68G01N33/53G01N33/574
Inventor 小园聪子秋山英雄妙本阳田中祥德郑基晚信正均野村修小川修中村英二郎辻本豪三
Owner TORAY IND INC
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