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Use of human phylaxin-1,2,3 protein in cancer detecting molecular mark

A technology of human defensin and detection label, applied in biological testing, material inspection products, etc., to reduce the cost of detection and diagnosis and improve the accuracy

Inactive Publication Date: 2007-04-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports about HNP-1, -2, -3 proteins as molecular markers for early detection and diagnosis of tumors

Method used

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  • Use of human phylaxin-1,2,3 protein in cancer detecting molecular mark
  • Use of human phylaxin-1,2,3 protein in cancer detecting molecular mark
  • Use of human phylaxin-1,2,3 protein in cancer detecting molecular mark

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 High performance liquid chromatography (HPLC) method to detect the expression difference of HNP-1, -2, -3 protein in tumor tissue and its adjacent normal tissue

[0017] 1. Experimental Materials and Instruments

[0018] 15 pairs of colorectal cancer and its adjacent normal colorectal tissues, 5 pairs of colorectal cancer metastasized lung cancer tissues and their adjacent normal lung tissues, 15 pairs of gastric cancer tissues and their adjacent normal gastric tissues, the above tissues were obtained from the Second Affiliated Hospital of Zhejiang University Acquired from the Tissue Bank of the Cancer Institute. Analytical pure acetocyanide (CAN), acetic acid, trifluoroacetic acid (TFA), liquid chromatograph (HPLC) is the HEWLETT PACKARD HP series 1100 product of U.S. Hewlett Packard Company, and the separation column of HPLC is the C18 column (2.0 ×150mm, 300A).

[0019] 2. Experimental method

[0020] Take the tissue sample out of liquid nitrogen and was...

Embodiment 2

[0027] Example 2 Identification of NP-1, -2, -3 proteins by surface-enhanced laser desorption ionization-time-of-flight-mass spectrometry (SELDI-TOF-MS)

[0028] 1. Experimental materials

[0029] CHCA; gold chip; SELDI-TOF-MS mass spectrometer (CIPHERGEN, USA), DTT (dithiothreitol), NP20 chip of All-in-one standard protein.

[0030] 2. Experimental method

[0031] Collect the above-mentioned characteristic absorption peaks obtained by HPLC analysis, that is, the elution peak at the 63% elution concentration of CAN, dissolve the protein with an appropriate amount of tissue lysate (20% acetonitrile and 2% acetic acid) after freeze-drying, Take 1 μl of protein solution and apply it to the sample hole of the gold chip, add 0.5 μl of CHCA surface enhancer after drying at room temperature, perform SELDI-TOF-MS analysis after room temperature drying and use Ciphergen proteinchip 3.1 software to read the data. Before data collection, the SELDI instrument was calibrated by the All-i...

Embodiment 3

[0037] Example 3 Comparison of HNP-1, -2, -3 protein differences in tumor tissue and its adjacent normal tissue by flight mass spectrometry SELDI-TOF-MS

[0038] 1. Experimental materials

[0039] CHCA, gold chip, SELDI-TOF-MS mass spectrometer (CIPHERGEN, USA), DTT, NP20 chip of All-in-one standard protein.

[0040] 2. Experimental method

[0041] Take the tissue protein solution extracted in Example 1, and adjust the concentration of each tissue protein solution to 2 μg / μl. 1 μl of the above protein solution was taken from each tissue sample for SELDI analysis, and the SELD analysis and data collection methods were the same as in Example 2.

[0042] 3. Experimental results

[0043] See Figure 3 for the comparison results of SELDI analysis. Figure 3A is the SELDI-TOF-MS analysis chart of protein in lysate of lung cancer tissue with gastric cancer, colorectal cancer and colorectal cancer metastasis; Figure 3B is the protein expression of normal tissue lysate in adjacent ti...

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Abstract

The present invention provides one kind of molecular marker for the early stage detection and diagnosis of carcinoma of large intestine and gastric cancer, and the molecular marker is human neutrophil phylaxin HNP-1, -2, -3 protein. The process of detecting HNP-1, -2, -3 protein in tumor tissue includes high performance liquid chromatography (HPLC), SELDI-TOF-MS and immuno-histochemistry. The present invention has the advantages of high accuracy in early stage detection of carcinoma of large intestine and gastric cancer and lowered tumor detecting and diagnosing cost.

Description

technical field [0001] The invention relates to a new molecular marker for cancer detection, and mainly relates to the application of human defensin-1, -2 and -3 proteins as molecular markers in cancer detection. Background technique [0002] Human neutrophil defensin HNP-α is a kind of arginine-rich positive polypeptide molecule, which consists of 29-33 amino acids. There are three intrachain disulfide bonds formed by 6 conserved cysteine ​​residues in the protein molecule, making the protein form a three-bundle sheet structure. Among the six defensin-α family members that have been discovered so far, HNP-1, -2, -3, and -4 are mainly expressed in neutrophils, while HD5 and HD6 are found in Paneth cells of the small intestine. However, studies have reported that HNP1-3 protein can also be expressed in some epithelial cells, specific lymphocytes and some tumor cells. [0003] Existing studies have proved that human neutrophil defensins HNP-1, -2, -3 proteins perform certain...

Claims

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Application Information

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IPC IPC(8): G01N33/52
Inventor 张昆胡汛
Owner ZHEJIANG UNIV
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