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PNC detection kit for detecting tumor invasion and application thereof

A detection kit and invasive technology are applied in the field of detection kits for the detection of PNC, a new target of tumor invasiveness, which can solve the problems of lack of diagnostic kits for tumor invasiveness and lack of standards and reference basis for individualized chemotherapy regimens. Throughput screening, simple method, good stability effect

Inactive Publication Date: 2013-10-16
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is still a lack of clear standards and references for the evaluation of tumor prognosis and the formulation of individualized chemotherapy regimens in clinical practice, and there is still a lack of tumor aggressiveness diagnostic kits suitable for clinical detection at home and abroad.

Method used

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  • PNC detection kit for detecting tumor invasion and application thereof
  • PNC detection kit for detecting tumor invasion and application thereof
  • PNC detection kit for detecting tumor invasion and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] 1. Collect tumor puncture specimens and paraffin-embedded sections of invasive ductal carcinoma of the breast at different clinical stages and pathological grades.

[0070] 2. Use PNC immunohistochemical kit for detection:

[0071] 1. After the paraffin sections are dewaxed and hydrated, wash them three times with PBS (pH=7.4), 3 minutes each time;

[0072] 2. Dilute the 10×antigen retrieval concentrate 10 times with ultrapure water, submerge the sectioned tissue, and boil in a microwave oven for 2 minutes to restore tissue antigens;

[0073] 3. Add 50 μl of peroxidase blocking solution dropwise to each slice and incubate at room temperature for 10 minutes to block the activity of endogenous peroxidase. Wash with PBS (pH=7.4) three times, 3 minutes each time;

[0074] 4. Remove the PBS solution, add 50 μl of blocking serum to each slice, and incubate at room temperature for 30 minutes;

[0075] 5. Remove the serum, add 50 μl of PTB antibody (1:200 dilution) (or drop ...

Embodiment 2

[0085] 1. Liver tissue microarray chip customization for liver cancer: including 30 cases of hepatocellular carcinoma, 12 cases of intrahepatic cholangiocarcinoma, 5 cases of metastatic cancer, 8 cases of liver cirrhosis, 10 cases of inflammation, 5 cases of paracancerous tissue, and 5 cases of normal liver tissue , one point per case.

[0086] 2. Use PNC immunohistochemical kit for detection:

[0087] 1. After the paraffin sections are dewaxed and hydrated, wash with PBS three times, each time for 3 minutes;

[0088] 2. Dilute the 10×antigen retrieval concentrate with PBS, submerge the sliced ​​tissue, and boil in a microwave oven for 2 minutes to restore tissue antigens;

[0089] 3. Add 50 μl peroxidase blocking solution dropwise to each slice and incubate at room temperature for 10 minutes to block the activity of endogenous peroxidase. Rinse with PBS three times, 3 minutes each time;

[0090] 4. Remove the PBS solution, add 50 μl blocking serum dropwise to the slice, an...

Embodiment 3

[0101] 1. The chemotherapeutic drug Amonafide, as a class of Topo II inhibitors, has the advantages of being highly effective and not causing drug resistance, but because it produces toxic metabolites that lead to bone marrow suppression and has relatively large toxic and side effects, further clinical research was terminated. A series of derivatives of Amonafide (A1~A10) were synthesized by modifying the structure of its benzene ring group.

[0102] 2. HepG2 cells were plated in a 24-well cell culture plate with ordinary cell culture medium (high glucose DMEM+10% FBS), 1ml / well.

[0103] 3. Amonafide derivative compounds A1~A10 were dissolved in DMSO respectively, and then serially diluted.

[0104] 4. After the cells adhered to the wall (overnight), the ordinary cell culture medium was discarded, and the cell culture medium containing gradient doses (1μM, 10μM, 30μM and 100μM) of Amonafide derivative compounds were added to each well, and DMSO was used as a control.

[0105...

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Abstract

The invention relates to the field of biotechnology, especially to a detection kit of a novel target PNC for detection of tumor invasion and application thereof. The first detection antibody of the PNC detection kit for detecting tumor invasion is a PTB or CUG-BP1 antibody. The invention describes composition and application methods of a PNC immunohistochemical detection kit and a PNC immunofluorescence detection kit used for detecting tumor invasion. Furthermore, the invention also provides a method for screening a novel anti-metastasis compound and a Chinese herbal medicine monomer thereof; and the method for screening the novel anti-metastasis compound is also applicable to screening of small molecule compounds and Chinese herbal medicine monomers thereof. According to the invention, the proportion of PNC in a tissue sample of a cancer patient obtained through paracentesis is detected by using the PNC kits, so detection is simple and convenient, is easy to operate and has good stability; the proportion of PNC in tumor cells which have been affected by the compound is detected by using the PNC kits, computer programming and analysis are cooperatively used, so high throughput screening of a novel drug is realized.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit for detecting PNC, a new tumor invasive target, and an application thereof. Background technique [0002] With the increasing rate of new malignant tumors worldwide, tumors have become one of the major diseases that endanger human health. The mortality rate of malignant tumors in my country also shows a continuous increase trend, and invasiveness (infiltration-metastasis), as one of the important signs of malignant tumors, is the main cause of death of cancer patients. Therefore, it is necessary to explore effective and easy-to-monitor tumor invasiveness. molecular markers. The mechanism of tumor invasiveness and the exploration of new molecular markers are currently a hot spot in the field of tumor biology research. However, at present, there is still a lack of clear standards and references for the evaluation of tumor prognosis and the formulation of individualized...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 陈智刘艳宁黄穗楼国华
Owner ZHEJIANG UNIV
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