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Immunohistochemical diagnosis kit for colorectal cancer malignancy and metastasis of colorectal cancer

A diagnostic kit and technology for colorectal cancer, which can be used in disease diagnosis, biological testing, material testing, etc., and can solve the problems of lack of experiments and research

Inactive Publication Date: 2014-01-01
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although domestic and foreign studies have shown that S100A8 is significantly highly expressed in gastric cancer, colon cancer, prostate cancer, and breast cancer, there is still a lack of more detailed information about the relationship between S100A8 protein and the occurrence and development of specific tumors, as well as the application of S100A8 in the diagnosis and treatment of specific tumors. In-depth experiments and research

Method used

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  • Immunohistochemical diagnosis kit for colorectal cancer malignancy and metastasis of colorectal cancer
  • Immunohistochemical diagnosis kit for colorectal cancer malignancy and metastasis of colorectal cancer
  • Immunohistochemical diagnosis kit for colorectal cancer malignancy and metastasis of colorectal cancer

Examples

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Effect test

Embodiment 1

[0023] Example 1 Preparation of Immunohistochemical Diagnostic Kit for Malignancy and Metastasis of Colorectal Cancer

[0024] The inventor used the NCBI high-throughput gene expression database GEO DataSets (gene expression omnibus) to analyze the S100A8 RNA level microarray data of 70 cases of colorectal cancer tissues and 12 cases of normal colorectal tissues in the series GSE9348. The results are shown in the attached figure 1 shown. Through this study, it was found that the expression level of S100A8 in colorectal cancer was significantly higher than that in normal colorectal tissue, p<0.05, and the difference was statistically significant. Further experiments by the inventors by immunohistochemistry showed that the expression level of S100A8 in colorectal cancer was significantly higher than that in the normal colorectal group, and the expression level of S100A8 protein was significantly different in colorectal cancer tissues with different degrees of differentiation, hi...

Embodiment 2

[0028] Example 2 Using the kit prepared in Example 1 to detect colorectal cancer tissue samples

[0029] (1) Dewaxing of baked slices: Take the tissues of colorectal cancer patients, put the tissue slices to be stained in a 60°C oven and bake for 60-90 minutes (depending on the quality of the slices), then quickly put them in xylene (I, II ) for 10 minutes × 2 times for dewaxing;

[0030] (2) Hydration: Place the slices in the following gradient alcohols in order: soak in 100% ethanol for 5 minutes - soak in 90% ethanol for 5 minutes - soak in 75% ethanol for 5 minutes - soak in 50% ethanol for 5 minutes -- soak in distilled water for 3 minutes -- wash with PBS for 3 minutes x 3 times;

[0031] (3) Antigen retrieval: Boil 0.01M citric acid antigen retrieval solution at pH6.0 in a microwave oven, place slices in it, seal the container mouth with latex gloves, continue heating in boiling water for 15-20 minutes, and keep the temperature at 100°C , After the container is natura...

Embodiment 3

[0041] Example 3 Using the kit of the present invention to detect colorectal cancer tissue samples for clinical verification

[0042] The expression of S100A8 protein in 54 cases of normal and 104 cases of colorectal cancer gross specimens was detected by using the kit of Example 1 and according to the method and steps of Example 2. All tissue samples were pathologically confirmed by the Pathology Department of the Second Xiangya Hospital of Central South University after surgery; 58 of the 104 colorectal tumor samples were non-metastatic cancers, and 46 were distant metastatic cancers; 96 of them could be used for malignancy analysis: There were 14 well-differentiated carcinomas, 48 ​​moderately differentiated carcinomas, and 34 poorly differentiated carcinomas.

[0043] According to the localization of the protein in the cell, a comprehensive score was made according to the two criteria of protein expression positive signal intensity and positive expression cell number. Cri...

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Abstract

The invention discloses an immunohistochemical diagnosis kit for colorectal cancer malignancy and metastasis of colorectal cancer. The kit comprises an endogenous peroxidase inhibitor, a non-specific binding site inhibitor, a primary antibody, a secondary antibody, an enzyme and a color developing agent. The immunohistochemical diagnosis kit is characterized in that the primary antibody is a human S100A8 monoclonal antibody or a murine S100A8 monoclonal antibody, and the secondary antibody is biotin-labeled goat anti-mouse IgG or rabbit anti-mouse IgG. According to the kit, expressions of S100A8 antigens on the surface of colorectal pathological tissues are detected, so that the colorectal cancer malignancy and metastasis conditions can be rapidly and accurately judged, and a help is provided for clinical diagnosis of the colorectal cancer.

Description

technical field [0001] The invention relates to a tumor immunodiagnostic kit, in particular to a colorectal cancer immunohistochemical kit. Background technique [0002] S100 proteins are a family of calcium-binding proteins consisting of small molecular weight proteins (10-14 kDa), named for their ability to dissolve in 100% saturation ammonium sulfate at neutral pH. Studies have found that such proteins are only expressed in vertebrates, and 23 members of the S100 protein family have been discovered so far. The S100A8 protein is one of the important members of the S100 protein family. It has the characteristic conserved EF hand structure and 4-helical ring of the S100 protein family. 2+ , Zn 2+ and Cu 2+ Structural changes occur in combination with different metal ions, exposing the binding site with the target protein and then participating in biological effects such as inflammatory response, immune response and tumor development by interacting with the corresponding e...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574
CPCG01N33/57419G01N33/57407G01N33/57446G01N33/68G01N2800/7028
Inventor 沈守荣马健张学梅李夏雨李楠唐岸柳
Owner CENT SOUTH UNIV
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