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Application of ADAM12m used as tumor marker and target molecule in diagnosing and treating tumors

A technology for tumors and uses, applied in the field of metastatic target proteins and targeted drugs, tumor invasion, and can solve problems such as lack of protein research

Inactive Publication Date: 2010-09-08
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been found that ADAM12m is up-regulated in tumor tissue and participates in tumor proliferation and apoptosis, but there is no research on the protein's tumor target

Method used

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  • Application of ADAM12m used as tumor marker and target molecule in diagnosing and treating tumors
  • Application of ADAM12m used as tumor marker and target molecule in diagnosing and treating tumors
  • Application of ADAM12m used as tumor marker and target molecule in diagnosing and treating tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation of embodiment 1 tumor antigen

[0040] According to the prior art, the cDNA coding sequence of the ADAM12m gene is cloned, constructed on the expression vector PET-30b, and the ADAM12m protein is expressed in prokaryotic cells.

[0041] The amino acid sequence of the expressed ADAM12m protein is:

[0042] H2N-MAARPLPVSPARALLLALAGALLAPCEARGVSLWNQGRADEVVSASVGSGDLWIPVKSFDSKNHPEVLNIRLQRESKELIINLERNEGLIASSFTETHYLQDGTDVSLARNYTVILGHCYYHGHVRGYSDSAVSLSTCSGLRGLIVFENESYVLEPMKSATNRYKLFPAKKLKSVRGSCGSHHNTPNLAAKNVFPPPSQTWARRHKRETLKATKYVELVIVADNREFQRQGKDLEKVKQRLIEIANHVDKFYRPLNIRIVLVGVEVWNDMDKCSVSQDPFTSLHEFLDWRKMKLLPRKSHDNAQLVSGVYFQGTTIGMAPIMSMCTADQSGGIVMDHSDNPLGAAVTLAHELGHNFGMNHDTLDRGCSCQMAVEKGGCIMNASTGYPFPMVFSSCSRKDLETSLEKGMGVCLFNLPEVRESFGGQKCGNRFVEEGEECDCGEPEECMNRCCNATTCTLKPDAVCAHGLCCEDCQLKPAGTACRDSSNSCDLPEFCTGASPHCPANVYLHDGHSCQDVDGYCYNGICQTHEQQCVTLWGPGAKPAPGICFERVNSAGDPYGNCGKVSKSSFAKCEMRDAKCGKIQCQGGASRPVIGTNAVSIETNIPLQQGGRILCRGTHVYLGDDMPDPGLVLAGTKCADGKICLNRQCQNISVFGVH...

Embodiment 2A

[0043] Example 2 Preparation of ADAM12m protein polyclonal antibody

[0044] In this study, the ADAM12m protein expressed in Example 1 was used to immunize rabbits to prepare polyclonal antibodies against ADAM12m protein. Pre-immunization serum: before immunization, blood was collected from the rabbit's ear vein for inspection. For the first immunization: ADAM12m protein and Freund's complete adjuvant were fully mixed at a ratio of 1:1, and then injected intradermally to immunize rabbits. 200 μg 383 polypeptide KLH conjugate. The second immunization: 14 days after the first immunization, take 200 μg of ADAM12m protein and Freund's incomplete adjuvant to fully mix, and then intradermally inject again to immunize rabbits. BCG injection: 5 days after the second immunization, the rabbits were immunized by multi-point subcutaneous injection of 50 mg BCG per rabbit. The third immunization: after 10 days from the day of BCG injection, the dose of 200 μg ADAM12m protein per rabbit w...

Embodiment 3

[0045] Example 3 The expression of ADAM12m protein in tumor tissue increases (tissue chip analysis)

[0046] Tissue chips were prepared from paraffin-embedded esophageal cancer tissues, routinely paraffin-sectioned, dewaxed with xylene, and hydrated with graded alcohol. 3% hydrogen peroxide at room temperature for 10 minutes to block the activity of endogenous peroxidase. Sections were subjected to antigen retrieval by microwave heating in citrate buffer (0.01M, pH 6.0). Block with 10% normal goat serum for 30 min at room temperature. Discard the blocking solution, add 1ug / ml anti-ADAM12m protein polyclonal antibody, and incubate overnight at 4°C. Then wash 3 times with PBS, 5min each time. Add an appropriate concentration of biotin-labeled goat anti-rabbit secondary antibody and incubate at room temperature for 30 min. Then wash with PBS 3 times, 5 min each time, add HRP-labeled streptavidin, and incubate at room temperature for 30 min. DAB staining, counterstaining with...

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Abstract

The invention relates to application of ADAM12m used as a tumor marker and a target molecule in diagnosing and treating tumors. The ADAM12m is used as a mark of tumor invasion metastasis and participates in the processes of growth and invasion metastasis of the tumors, thus the ADAM12m can be used as a target spot in cancer diagnosis and / or treatment and for screening medicaments for resisting the tumor invasion metastasis.

Description

technical field [0001] The present invention relates to tumor invasion, metastasis target protein and targeted drug. Specifically, the present invention relates to a novel tumor invasion and metastasis targeting protein ADAM12m and its use in screening and / or preparing drugs for treating and diagnosing tumor invasion and metastasis. The invention also relates to the specific antibody of the protein and its application. Background technique [0002] Tumor is one of the most common human diseases, and its death rate ranks second and third among various diseases in the world and our country, respectively, seriously affecting human health. With the development of society in our country, the incidence of tumor is still increasing year by year. There are currently about 4.5 million cancer patients in my country, with a mortality rate of over 30%, and the number of new patients is as high as 2 million or more each year. Effectively treating cancer has become a top priority in sc...

Claims

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Application Information

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IPC IPC(8): G01N33/68A61K39/395A61K31/7088A61P35/00A61P35/04
Inventor 杨治华冉宇靓胡海
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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