Method for detecting gynecological cancer

An ovarian cancer and antibody technology, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, scientific instruments, etc., can solve the correlation between β3Gal-T5 and the correlation between ovarian cancer and endometrial cancer that has not been reported. and other problems, to achieve the effect of increasing the detection rate and achieving a significant effect.

Inactive Publication Date: 2015-03-11
LSIP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, there is no report showing the correlation between β3Gal-T5 and ovarian cancer and uterine body cancer, and the correlation between GlcNAc6ST-2 and uterine body cancer, and the β1-3 hemitransferase, one of the β1-3 galactosyltransferase family Lactose transferase-4 (hereinafter referred to as β3Gal-T4), and its correlation with ovarian cancer and uterine body cancer has not been reported

Method used

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  • Method for detecting gynecological cancer
  • Method for detecting gynecological cancer
  • Method for detecting gynecological cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] "Example 1: Preparation of anti-β3Gal-T5 antibody"

[0111] (A) Preparation of soluble β3Gal-T5 protein (preparation of antigen for immunization)

[0112] (A-1) Construction of β3Gal-T5 expression vector

[0113] A cDNA fragment encoding a peptide lacking cytoplasmic and transmembrane regions was amplified from a human large intestine cDNA library by PCR using the b3Gal-T5 forward primer and b3Gal-T5 reverse primer described below. The obtained cDNA fragment was a cDNA encoding a peptide containing the 27th to 310th amino acid sequence of β3Gal-T5.

[0114] Forward primer for b3Gal-T5: 5'-tttgtcgacAGTCTAAATCCTTTCAAAG-3' (SEQ ID NO: 1)

[0115] Reverse primer for b3Gal-T5: 5'-tttaagcttCAGACAGGCGGACAATC-3' (SEQ ID NO: 2)

[0116] Among the sequences, the sequence described in lowercase letters is a sequence containing a restriction enzyme site for ligation with an expression vector. The obtained cDNA was cloned into the cloning site of the pQE9 vector (Kiagen Corporat...

Embodiment 2

[0140] "Example 2: Expression of β3Gal-T4 and β3Gal-T5 mRNA in Ovarian Cancer Cells"

[0141] Expression of β3Gal-T4 and β3Gal-T5 mRNA in ovarian cancer cells was investigated by RT-PCR using five ovarian cancer cell lines. ES-2 cells (clear cell carcinoma of the ovary) were purchased from ATCC, MCAS cells (mucinous cystadenocarcinoma of the ovary), RMG-I cells (mesonephric adenocarcinoma of the ovary), RMG-II cells (mesonephric adenocarcinoma of the ovary), and RMUG-L (mucinous cystadenocarcinoma of the ovary) was purchased from the Human Science Research Resource Bank (Hydmancers Research Resource Bank).

[0142] The collected cells were lysed using ISOGEN (Nipponce Incorporated), and total RNA was isolated. cDNA was synthesized from the obtained RNA using oligo dT primer and SuperScript III (Invitrogen). The cDNA amount was corrected by that of β-actin.

[0143] Expression of β3Gal-T4 and β3Gal-T5 was confirmed by PCR after adding the following primers and incubating at ...

Embodiment 3

[0150] "Example 3: Preparation of GlcNAc6ST-2 Antibody"

[0151] (A) Preparation of soluble GlcNAc6ST-2 protein (preparation of antigen for immunization)

[0152] (A-1) Construction of GlcNAc6ST-2 expression vector

[0153] The cDNA encoding the peptide that lacks the cytoplasmic and transmembrane regions was amplified from the previously obtained full-length cDNA (Glycobiology, 2002, 12, pages 379-388) by PCR using the following GlcNAc6ST2 forward primer and GlcNAc6ST2 reverse primer fragment. The obtained cDNA fragment is a cDNA encoding a peptide containing the 28th to 386th amino acid sequence of GlcNAc6ST-2.

[0154] Forward primer for GlcNAc6ST2: 5'-tttgtcgacAGCCACAACATCAGCT-3' (SEQ ID NO: 17)

[0155] Reverse primer for GlcNAc6ST2: 5'-tttaagcttAGTGGATTTGCTCAG-3' (SEQ ID NO: 18)

[0156] Among the sequences, the sequence described in lowercase letters is a sequence containing a restriction enzyme site for ligation with an expression vector. The obtained cDNA was clo...

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Abstract

The object of the present invention is to provide a method for detecting gynecologic cancer and a kit for detecting the same. The object can be solved by a method for detecting gynecologic cancer characterized in that ²1,3-galactosyltransferase-5 and / or ²1,3-galactosyltransferase-4, and GlcNAc 6-O-sulfotransferase-2 are analyzed. According to the detecting method, ovarian cancer and endometrial cancer patients can be detected at an early stage i.e. stages I and II, wherein patients have no subjective symptoms.

Description

technical field [0001] The invention relates to a detection method for gynecological cancer, especially ovarian cancer or uterine body cancer, which is characterized in that: β1-3 galactosyltransferase-5 and / or β1-3 galactosyltransferase-4, or N-acetyl Glucosamine 6-O-sulfotransferase-2 was analyzed. The above-mentioned "analysis" in this specification includes both "measurement" for quantitatively or semi-quantitatively determining the amount of an analyte, and "detection" for determining the presence or absence of an analyte. Background technique [0002] Gynecological cancers include cervical cancer, uterine body cancer, ovarian cancer, vulvar cancer, vaginal cancer, uterine sarcoma, choriocarcinoma (villous disease), etc. Most of them are difficult to detect early by symptoms, and the 5-year survival rate is significantly increased by surgical treatment such as surgery in the early stage. Therefore, development of tumor markers for early detection of these gynecologica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/573C07K16/40
CPCG01N33/57442C12Q1/48C07K16/40G01N33/57449G01N2333/91097
Inventor 山下克子濑古玲青木大辅坂本优
Owner LSIP
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