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Human colorectal cancer protein mark CLCA1 (calcium-activated chloride channel 1) kit and application thereof

A colorectal cancer and kit technology, applied in the fields of biotechnology and medicine, can solve the problems of large interference in pathological judgment, low prediction specificity, etc., and achieve the effect of low cost

Inactive Publication Date: 2017-08-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Secondly, conventional methods rely on factors such as pathological parameters and TNM staging to predict the prognosis and survival of patients after treatment. High, specific, and easy-to-operate molecular markers are used as auxiliary judgment methods

Method used

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  • Human colorectal cancer protein mark CLCA1 (calcium-activated chloride channel 1) kit and application thereof
  • Human colorectal cancer protein mark CLCA1 (calcium-activated chloride channel 1) kit and application thereof
  • Human colorectal cancer protein mark CLCA1 (calcium-activated chloride channel 1) kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Preparation of CLCA1 immunohistochemical detection kit

[0023] The kit includes specific antibodies against CLCA1, EDTA antigen retrieval solution, endogenous peroxidase blocking agent, goat serum, secondary antibody, PBS buffer, hematoxylin, differentiation solution, DAB chromogenic solution,

[0024] Wherein the endogenous peroxidase blocking agent is 3% hydrogen peroxide;

[0025] PBS buffer preparation method: weigh 8.0g NaCl, 0.2g KCl, 1.44g Na2HPO4, 0.24g KH2PO4, add distilled water to 1000ml, mix well, and adjust the pH to 7.2-7.6;

[0026] EDTA Antigen Retrieval Liquid Formula: Weigh 30.27g of Tris, 1.461g of EDTA (ethylenediaminetetraacetic acid), add distilled water to 500ml, mix well, and dilute 10 times before use;

[0027] Hematoxylin preparation method: weigh hematoxylin 2g, dissolve in 250mL absolute ethanol; weigh aluminum sulfate 17.6g, dissolve in 750mL distilled water; mix the dissolved hematoxylin and aluminum sulfate, stir with a glass r...

Embodiment 2

[0030] Example 2 detects the sample to be tested with the prepared kit

[0031] The operation process is as follows:

[0032] 1. Paraffin section: The surgical specimens diagnosed by pathology as colorectal cancer and their adjacent normal tissues were fixed in formalin and embedded in paraffin to make paraffin blocks. The thickness of the slices was 5 microns. Receive antitumor therapy.

[0033] 2. Bake slices: Put the paraffin slices in a 60-degree Celsius incubator and bake the slices for 2 hours.

[0034] 3. Dewaxing and Hydration:

[0035] 1) Put the paraffin sections into two cylinders of xylene in turn, dewax twice, each time for 20 minutes;

[0036] 2) Hydration: Put the paraffin sections into two vats of absolute ethanol in turn, 5 minutes each; take out the paraffin sections, and place them in two vats of 95% ethanol, 5 minutes each; take out the paraffin sections, and put them in 90% ethanol in ethanol for 5 minutes; take out the paraffin sections and put them i...

Embodiment 3

[0056] The clinical verification of embodiment 3 kits

[0057] Using the kit prepared in Example 1 above, according to the detection steps and method in Example 2, the expression of CLCA1 in 18 pairs of colorectal cancer tissues and their paracancerous tissues was detected. All tissue samples were pathologically confirmed by the Pathology Department of the Second Affiliated Hospital of Zhejiang University School of Medicine after surgery. Randomly select at least 10 high-magnification fields of view (×200) from each slice, count at least 1000 cells, and calculate the results with the integral method. That is, joint scoring was performed according to the staining intensity and positive cell ratio of each section. Staining intensity: 0 points for colorless; 1 point for light yellow; 2 points for brownish yellow; 3 points for brown. Positive cell ratio: 50% is 3 points. Take the average of the sum of the two scores of the staining intensity and the ratio of positive cells, tho...

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Abstract

The invention provides a human colorectal cancer protein mark CLCA1 (calcium-activated chloride channel 1) kit, which comprises a specific antibody aiming at CLCA1, an EDTA (ethylene diamine tetraacetic acid) antigen restoration solution, an endogenous peroxidase blocker, goat serum, second antibodies, a PBS buffer solution, hematoxylin and a DAB color developing solution. Study results show that the expression quantity of the CLCA1 in the normal colorectal tissues is obviously higher than that in the colorectal cancer tissues; the content of the CLCA1 in the health people blood is obviously higher than the content in the blood of the colorectal cancer patient; in addition, the lifetime of the colorectal cancer patients with high CLCA1 expression is obviously longer than the patient with low CLCA1 expression; the scientific basis is provided for effectively judging the colorectal cancer canceration progress, detecting the human colorectal cancer in the early stage and judging the prognosis. Therefore the kit provided by the invention is used for preparing a preparation for screening, diagnosing or predicting the human colorectal cancer, can be used for screening anti-colorectal-cancer medicine targets, and are also used for preparing medicine for relieving or treating the human colorectal cancer. The kit provided by the invention is efficient and convenient, and the cost is lower.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and relates to a human colorectal cancer marker kit, in particular to a human colorectal cancer marker CLCA1 kit, which is a protein calcium ion-activated chloride channel protein 1 (Calcium-activated chloride channel 1, CLCA1), and the application of the kit. Background technique [0002] According to the latest statistics from the World Health Organization, globally, the incidence of colorectal cancer ranks third among all malignant tumors, and its mortality rate ranks fourth. The early symptoms of colorectal cancer are not obvious, and so far, there is no marker with good specificity and sensitivity for early detection of colorectal cancer, and more than half of the patients are locally advanced or advanced when diagnosed. Clinical stage is an extremely important factor in determining the prognosis of patients with colorectal cancer. The 5-year survival rate of early-stage patients ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574G01N33/531
CPCG01N33/68G01N33/531G01N33/57407G01N2800/065
Inventor 李晓芬余捷凯胡望雄彭佳萍袁瑛郑树
Owner ZHEJIANG UNIV
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