CN107929732A - The application of CD146 and its antibody diagnosis and treatment triple negative breast cancer - Google Patents
The application of CD146 and its antibody diagnosis and treatment triple negative breast cancer Download PDFInfo
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Abstract
The present invention relates to the application of CD146 and its antibody diagnosis and treatment triple negative breast cancer.Present invention firstly provides the new target drone that CD146 is triple negative breast cancer, anti-CD146 antibody is likely to become the novel targeted medicine for treating this kind of disease.Therefore, the present invention provides the antibody of CD146 or anti-CD146 or the functional form of the antibody and is preparing the application in being used to diagnose and/or treat the medicine of triple negative breast cancer.CD146 molecules specificity overexpression in triple negative breast cancer tissue, and induce tumour cell that epithelial cell occurs to transition of mesenchymal cells, promote tumor invasion migration.Therefore, the mechanism of action of CD146 Antybody therapies triple negative breast cancer is mainly to suppress the interstitial cell feature of triple negative breast cancer cell, reduces it and shifts invasive ability.Compared with conventional chemotherapeutic drugs, the Small side effects of anti-CD146 antibody, target is clear and definite, will not cause the side effect of systemic body-wide.
Description
The application is application number 201110412778.X, denomination of invention " CD146 and its antibody diagnosis and the negative breast for the treatment of three
The divisional application of the application of gland cancer ".
Technical field
The invention belongs to immunodiagnosis and biological therapy field.Specifically, the present invention relates to CD146 molecules and its antibody
In the application of diagnose and treat breast cancer.More particularly it relates to CD146 monoclonal antibodies AA4 can to diagnose three cloudy
Property breast cancer, and the function of CD146 monoclonal antibodies AA98 and its functional form be suppress triple negative breast cancer cytoplasm
Cell characteristic and malignant characteristics, so as to suppress metastases invasion and attack.
Background technology
Breast cancer is the highest malignant tumour of most common in women and lethality.It is global to have nearly 1,300,000 women trouble every year
Upper breast cancer, and it is dead because of the transfer and relapse of breast cancer to have more than 400,000 women.Grade malignancy is highest in breast cancer
A kind of hypotype, is known as triple negative breast cancer, with estrogen receptor (ER) feminine gender/corpus luteum hormone acceptor (PR) feminine gender/epidermal growth
Factor acceptor (HER2) feminine gender is characterized.The incidence of triple negative breast cancer accounts for 17%~25% in breast cancer, due to lacking
Transfer and relapse occurs for the target of weary antiestrogenic and anti-HER2 targeted therapies, and high probability, is the worst one kind of prognosis
Breast cancer type.
At present, clinically treatment triple negative breast cancer is mainly chemotherapeutics, such as cis-platinum and anthracene nucleus medicament, mainly
Play the function of prevention metastases.This kind of medicine does not have targeting, will cause damage to the normal histoorgan of whole body yet, tool
There is very strong toxic side effect.In addition, although triple negative breast cancer chemotherapy early stage it is more sensitive to chemotherapeutics, but prognosis according to
It is old very bad.Therefore, the pathogenesis of triple negative breast cancer is explored, and therefrom excavates new target drone, is triple negative breast cancer treatment
New direction.
Lot of documents report, mechanism (epithelial to mesenchymal of the epithelial cell to transition of mesenchymal cells
Transition, EMT) play a significant role in the transfer and relapse of triple negative breast cancer.EMT assigns tumor cell invasion transfer
Ability, drug resistance and cells and characteristic of stem.For Genotyping, triple negative breast cancer belongs to the type of substrate sample breast cancer,
Its tumour cell is more likely to the mark molecule of expression interstitial cell, it is considered to be and one kind has occurred and that the breast cancer type of EMT,
And distal migration is occurred to important organs such as brain, lungs by the mechanism of EMT.Since the interstitial of triple negative breast cancer cell is thin
Born of the same parents' nature and characteristic explains the pernicious metastasis degree of tumour to a certain extent, suppresses its interstitial cell feature and excavates new phase
Target molecules are closed by as the potential direction of triple negative breast cancer cell therapy.Since many important growth factor receptorses are related
Tyrosine kinase activation, TGF-β associated signal paths, and the relevant signal path of the cell adhesion molecule such as integrin luring
The tumour cell for leading epithelial origin occurs to play a significant role during EMT, the antibody medicine using these signal paths as target
Thing, micromolecular inhibitor and antagonist are largely proved to reach the effect that treatment metastases are attacked by suppressing EMT
(Targeted therapies in control of EMT in carcinoma and fibrosis.Kian-Ngiap
Chua,Jing Ma,Jean-Paul Thiery.DOI:10.1016/j.ddmec.2008.06.002).For example, cystatin
C, a kind of serine inhibitor, being found in breast cancer cell can be as the antagonist of TGF-β acceptor, and then suppresses TGF-
The signal path that the combination of β and its acceptor and activation downstream induction EMT occur.Therefore, cystatin C can be used as potential
Medicine is used in the breast cancer treatment by TGF-β adjusting.
The content of the invention
Present invention firstly provides the new target drone that CD146 is triple negative breast cancer, anti-CD146 antibody is this kind of disease for the treatment of
Novel targeted medicine.Compared with traditional chemotherapeutics, the Small side effects of anti-CD146 antibody, will not cause the group of systemic body-wide
Knit organ damage.The application of CD146 and its antibody in triple negative breast cancer diagnose and treat is based on following important science
It was found that：(1) CD146 molecules specificity overexpression in triple negative breast cancer pathological tissue；(2) CD146 is in epithelial character mammary gland
After being overexpressed in cancer cell MCF-7 cells, EMT occurs for inducing cell, assigns cell invasion transfer ability；(3) it is newborn in the original location
The overexpression of CD146 molecules promotes mammary gland in gland cancer mouse models (Orthotopic Breast Cancer Mouse Model)
The formation of tumour, reduces the differentiation degree of tumour, and promotes invasion and attack in situ and the distal migration of tumour；(4) in three negative breasts
The expression of CD146 molecules or the functional antibodies AA98 using CD146 are lowered using siRNA in adenocarcinoma cell MDA-MB-231,
It can suppress its interstitial cell nature and characteristic, reduce cell transfer invasive ability.
In conclusion the overexpression of CD146 can induce the tumour cell of epithelial character that EMT occurs, this is to three negative breasts
The generation of gland cancer and progress play an important roll.Meanwhile CD146 molecules to maintain triple negative breast cancer interstitial cell feature and
Grade malignancy has critical function.The anti-CD146 functional antibodies AA98 of our independent developments can suppress three negative breasts in vitro
The migration of adenocarcinoma cell MDA-MB-231 and invasive ability.Therefore, it is proposed that CD146 can be as the target of triple negative breast cancer
Molecule is marked, its functional antibodies AA98 can be led to by the CD146 molecules of combination cell film surface by suppressing signal downstream
Road, so as to block it to induce the function of EMT, or suppresses the interstitial nature and characteristic of malignant cell.Meanwhile this kind of antibody
It is specific preferable, the damage of the immune function of systemic body-wide will not be caused.
Therefore, one aspect of the present invention is related to CD146 and is preparing for diagnosing and/or treating in triple negative breast cancer medicine
Using.Those skilled in the art can be directed to CD146 or its signal path by disclosed various method productions in the prior art
Inhibitor (such as antibody, siRNA etc.) so that prepare and be used to diagnose and/or treat the medicine of triple negative breast cancer.Diagnosing
During breast cancer, this aspect of the present invention includes being used for the various methods for detecting CD146 expression quantity, such as can detect the molecule in breast
Rna level or protein level in adenocarcinoma cell, and then auxiliary diagnosis triple negative breast cancer.
Further aspect of the present invention be related to the antibody of anti-CD146 or the functional form of the antibody prepare for diagnose and/or
Treat the application in triple negative breast cancer medicine.
In the present invention, the antibody of anti-CD146 or the functional form of the antibody may include anti-CD146 monoclonal antibody,
Genetic engineering antibody and humanized antibody.This antibody-like can largely disclosed document be easily prepared according to this area.
In the present invention, the antibody of anti-CD146 or the functional form of the antibody are the Function Couplings of the antibody of anti-CD146
Thing, wherein can be medicine, toxin, radioactivity agent, enzyme, cell factor or another antibody with the material of antibody binding.
In the present invention, the antibody of anti-CD146 is preferably monoclonal antibody AA98 and AA4.
Another aspect of the invention is related to the antibody of CD146 or anti-CD146 or the functional form of the antibody and is preparing diagnosis three
Application in negative breast cancer drug, it is characterised in that the level of CD146 in the section of detection subject tumor tissue pathology.At this
This aspect of invention, medicine can be prepared into the form of kit.
Another aspect of the invention is related to the antibody of anti-CD146 or the functional form of the antibody and is preparing for three negative targetings
Property treatment medicine in application.Preferably, the functional form of the antibody of the anti-CD146 or the antibody includes anti-CD146's
Monoclonal antibody, genetic engineering antibody and humanized antibody.It is highly preferred that the antibody of the anti-CD146 is monoclonal antibody
AA98 and AA4.
Antibody A A98, AA4, AA1 for being used in this specification etc. can be according to Chinese Patent Application No. 99107586.2, China
The description of number of patent application 200810057260.7 obtains.
In order to be more fully understood and with the application of the invention, provide the following example.
Brief description of the drawings
From detailed description below in conjunction with the accompanying drawings, features described above of the invention and advantage will be apparent from, wherein：
Fig. 1 .A：Clinical samples:CD146 specificity overexpressions in triple negative breast cancer patient tumor tissue pathology cuts into slices；
B：The non-triple negative breast cancers of triple negative breast cancer vs.：88.9%vs.8.7%, p<0.0001.
Fig. 2 .CD146 are overexpressed in MCF-7 cells, filter out stable clone.Wherein Mock is control, is transferred to zero load；
B10 and A5 is CD146 expression strains, and B10 medium expression CD146, A5 are high expression CD146.
EMT occurs for the overexpression inducing cell of Fig. 3 .CD146.It is relevant thin that EMT occurs successively for A-B, Mock, B10 and A5
Born of the same parents' form and signaling molecule change；C, Mock, B10 and A5 cell migration ability strengthen successively；D, Mock, B10 and A5 invasion and attack are thin
Born of the same parents' number.
Fig. 4 are horizontal in vivo, and CD146, which is overexpressed, to be caused to increase breast cancer tumour generation (A), and lowers Tumor Differentiation journey
Spend (B), and the migration (C) in situ of tumour；
Fig. 5 are horizontal in vivo, and CD146, which is overexpressed, promotes breast cancer distal migration (A-D)；
Fig. 6 lower CD146 expression in triple negative breast cancer cell line MDA-MB-231 and reverse its interstitial cell feature.
(A) Western Blot identify breast cancer cell line MCF-7, MDA-MB-231 and Hs78T cell EMT associated tagged moleculars
(B) change for causing EMT associated tagged moleculars is lowered in CD146 expression；(C) reverse for causing cellular morphology is lowered in CD146 expression；
(D-E) CD146 expression, which is lowered, causes cell migration and the decline of invasive ability；
The interstitial cell that the anti-CD146 functional antibodies AA98 of Fig. 7 suppress triple negative breast cancer cell line MDA-MB-231 is special
Levy (A-D).
Fig. 8 .Sigma companies eukaryon expression plasmid p3XFLAG-HA-cmv-14 collection of illustrative plates.
Embodiment
Embodiment one：CD146 specificity overexpressions in triple negative breast cancer patient tumor tissue pathology cuts into slices；
Cell adhesion molecule plays a significant role during the performance of mammary gland carcinogenesis.CD146 molecules are a related blood
The important cells adhesion molecule of pipe new life and metastases.In order to study effects of the CD146 in breast cancer, we are to 90 breasts
Adenocarcinoma tissue pathological section carries out immunohistochemical analysis, finds CD146 molecules specificity overexpression in triple negative breast cancer.
Main material：Breast cancer patients tumor tissues paraffin section, including 44 triple negative breast cancers and 46 non-three the moon
Property breast cancer tissue, derives from Tianjin tumour hospital；
Main agents：4% paraformaldehyde, dimethylbenzene, absolute ethyl alcohol, absolute methanol, hydrogen peroxide, PBS, mouse resist
CD146 monoclonal antibodies AA4；Antibody A A98, AA4, AA1 for being used in this specification etc. can be according to Chinese Patent Application No.
99107586.2, the description of Chinese Patent Application No. 200810057260.7 obtains.
Main method：Paraffin section makes：With 4% paraformaldehyde fix flesh tissue 24 it is small when after, paraffin embedding, below
For the immunohistochemistry process of paraffin section：
1) slice, thin piece is taken out, enters 37 DEG C of dewaxings of xylene solution twice, 30 minutes every time；
2) aquation in absolute ethyl alcohol × 2-95%-80%-70%-50%-30% and distilled water, 5 points every time of room temperature are entered
Clock；
3) 0.3% hydrogen peroxide/methanol solution, 37 DEG C of lucifuges are handled 30 minutes, eliminate the work of endogenous peroxydase
Property, PBS is washed three times；
4) pH6.0 citric acids are repaired the antigen hot repair of water-bath 30 minutes of 100 DEG C of liquid and are answered, natural cooling；
5) when 5% normal sheep serum, 37 DEG C of closings 1 are small；
6) the diluted primary antibodies of PBS (rabbit-anti CD146 resists more or AA4), 4 DEG C of overnight incubations are added；
7) PBS is washed three times；Anti- rabbit-the biotin of goat or goat-anti-mouse-biotin secondary antibody (being purchased from Zhong Shan Golden Bridge)
(1：1000) when 37 DEG C of incubations 1 are small, PBS is washed three times；
8) Avidin-HRP (1：1000) it is incubated 45 minutes at 37 DEG C；
9) developed the color 2-7 minutes with the DAB lucifuges now matched somebody with somebody, then carry out haematoxylin and redye.
10) it is dehydrated step by step：50-70-80-90-100-100% ethanol-xylenes, dry, resinene mounting.
11) make film in micro imaging system.
The results show that as shown in Figure 1, compared to non-triple negative breast cancer tissue, CD146 molecules are in triple negative breast cancer group
Knit middle specificity overexpression.CD146 expresses the cell membrane in tumour cell, and is more likely to expression at the edge of tumor migration.
Embodiment two：CD146 is overexpressed induction epithelial character breast cancer cell MCF-7 and EMT occurs；
The cell line mcf-7 cell (being purchased from Shanghai cell bank, catalog number (Cat.No.) TCHu74) that we have selected CD146 feminine genders is made
For cell model, it is found that CD146 high expression in pernicious triple negative breast cancer tissue is of great significance.MCF-7 is one
The breast cancer tumor cells system of a epithelial character, grade malignancy is low, and differentiation degree is high, and cell migration ability is extremely weak.We
CD146 molecules are overexpressed in MCF-7, builds and obtains 2 stably expressing cell lines, B10 and A5, the medium expression CD146 of wherein B10,
A5 is high expression CD146.Meanwhile we are also transferred to empty carrier in MCF-7 cells, construct control and stablize strain Mock.Specifically
Process is as detailed below.
Main agents：CD146cDNA(SEQ ID NO:7) it is initially Munich, France university Judith P.Johnson religions
Present is awarded, can also be bought at home by Wuhan Sanying Bio-Technology Co., Ltd. obtain (numbering at present：BC056418, carries
Constitution grain is pCMV-SPORT6), encoded protein sequence is shown in SEQ ID NO:8.We have selected eukaryon expression plasmid
P3XFLAG-HA-cmv-14 (buying in Sigma companies, numbering E4901) builds CD146 eukaryon expression plasmids, original plasmid figure
Spectrum is shown in Fig. 8.CD146cDNA is inserted into p3XFLAF-HA-cmv-14 by two restriction enzyme sites of EcoRI and BamHI, gained after structure
Purpose plasmid preserved for a long time by this laboratory.PCR goes out 5 ' and 3 ' end difference wherein from the vector plasmid containing CD146cDNA
CD146cDNA fragments with EcoRI and BamHI with primer be：5 '-CGG AAT TCA ATG GGG CTT of forward direction
CCC AGG CTG-3’(SEQ ID NO:1)；Reverse 5 '-GCG GAT TCA TGC CTC AGA TCG ATG TA-3 '
(SEQID NO:2).Transfection reagent is Roche Fugene HD.G418 reagents are bought in Sigma.Transwell technologies use
96 orifice plates (8m) are bought in U.S. CORNING (Coring).Antibody A A98, AA4, AA1 for being used in this specification etc. can be according to China
The description of number of patent application 99107586.2, Chinese Patent Application No. 200810057260.7 obtains.Anti- E-cadherin antibody,
Anti- cytokeratin antibody, anti-Fibronectin antibody, the purchase of anti-GAPDH antibody are in Abcam, anti-Vimentin antibody purchase
In Sigma.Pancreatin is bought in Gibco.Matrigel is purchased from BD biosciences companies.
Main method：Go out monoclonal using lipofectamine transfecting eukaryotic cells, and using G418 resistance screenings, then
The characteristic of gained cell is identified with Western Blot, flow cytometer, Transwell technologies etc..
(1) by MCF-7 cells kind in the Tissue Culture Dish of 2 35mm, reach cell density during transfection in second day
80% or so.
(2) according to the explanation of Roche Fugen HD transfection reagents, the MCF-7 cells in two 25mm culture dishes are turned respectively
P3XFlag plasmids or p3XFlag-CD146 plasmids are contaminated, being separately added into G418 (0.5g/ml) after 48h is screened；
After (3) 1 weeks, after waiting the cell of survival to cover with, after trypsin digestion cell, with fresh culture diluting cells density extremely
1X104A/ml, is planted in 96 orifice plates, to polyclonal progress monoclonal.
After the cell of (4) 96 orifice plates covers with, it is expanded to 35mm culture dishes and is cultivated, and using the method pair of flow cytometer showed
The expression quantity of CD146 is detected, and selects the CD146 clones of variable expression.
(5) clone that picks out is continued to cultivate about 2-3 months in the presence of G418, can be into after clone's stabilization
The experiments such as the morphologic observation of row subsequent cell, cell migration invasion and attack test.Tested for cell migration, 5X103A cell kind in
Upper hole in Tranwell plates, is cultivated in the MEM culture mediums of 50l serum-frees, is the MEM of 10% serum containing 200l in lower opening
Culture medium is as inducing, and after 14-18h, the culture medium in upper hole is blotted, and the cell being covered on film has been dipped in 70% alcohol
Cotton-ball bar wipe clean, film is taken off and is tipped upside down on slide.Migrate the cell to come and fix simultaneously crystal violet with 4% PFA
Dyeing, microscope are taken pictures and are counted.Each cell needs to design 3 parallel control holes.Test, needed on film pre- for cell invasion
First cover the Matrigel glue of 2.5mg/ml, 1X104A cell will be used for the experiment.Connect lower step with it is upper same.
As shown in figure 3, during in MCF-7 stable clone strains Mock, B10 and A5, this is a series of, it is observed that related EMT
Associated change.First, Mock cells maintain the form of the typical epithelial character cell of female parent MCF-7 cells, and cell is given birth in flakes
It is long, in paving stone shape, its close connection.However, greatly change, cell point occur for the A5 cellular morphologies of high expression CD146 molecules
It is scattered to grow up to shuttle.The B10 cellular morphologies of medium expression CD146 molecules are in two kinds of intermediateness.Secondly, Western
Blot experiments show that Mock cells express the mark molecule of typical epithelial cell, such as E-cadherin and Cytokeratin.
And A5 cells express the mark molecule of typical interstitial cell, such as Vimentin and Fibronectin.(Epithelial-
Mesenchymal Transitions in Development and Disease, Jean Paul Thiery, Herv é
Acloque, Ruby Y.J.Huang and M.Angela Nieto.Cell 139, November 25,2009).It is most important
It is, using Transwell technologies (Leukocyte transmigration through vascular endothelium.An
in vitro method.Allavena P,Del Maschio A.Methods Mol Biol.1999；96:171-6), we
It can find that high expression of the CD146 in A5 cells significantly enhances cell migration and invasive ability.
Embodiment three：CD146 is overexpressed the formation for promoting tumor of breast in breast cancer mouse models in the original location, reduces tumour
Differentiation degree, and promote invasion and attack in situ and the distal migration of tumour；
We construct the breast cancer mouse model of orthotopic transplantation using SCID/Beige, find horizontal CD146 in vivo
Occurrence and development of the high expression to breast cancer play an important roll.
Experimental method：SCID/Beige (being purchased from dimension tonneau China) mouse of 20 4 weeks size females is selected, is randomly divided into 2
Group.Respectively 1 × 10 is injected in mammary fat tissue pad6Clone's Mock or A5 cell of a MCF-7 cell deriveds is resuspended in 50ul
PBS, mixes with the Matrigel of 50ul 2.5mg/ml.Observe 1 time weekly, length and width and height are measured, to calculate gross tumor volume.
After 10 weeks, after thering is mice with tumor to become weakness, take off cervical approach and put to death all mouse, peel off tumour and important organ, 4%PFA are fixed
24 it is small when after, paraffin embedding, cuts into slices and does immunohistochemical analysis.
As a result as shown in Figure 4, Figure 5, Mock cells Tumor formation on SCID/Beige mouse is poor, there was only 3 in 10 mouse
Only into knurl, and gross tumor volume is less than normal, tumor tissues structural integrity, shows that differentiation degree is high, and without generation invasion and attack in situ and far
End migration.And sharp contrast is formed, the A5 cells of CD146 high expression are easy into knurl, there was only 8 in 10 mouse into knurl,
And gross tumor volume is generally bigger than normal, tumor tissues structure is chaotic, shows that differentiation degree is low.Importantly, the tumour that A5 is formed is shown
Write ground invasion and attack and arrive neighbouring normal structure, such as skin, mammary gland, rib cage etc., and be transferred to lung and liver.
Example IV：CD146 or anti-CD146 functional antibodies are lowered in triple negative breast cancer cell line MDA-MB-231
Its interstitial cell feature of AA98 expression inhibitings.
Main agents：Target the siRNA sequence of CD146：5 '-CCA GCU CCG CGU CUA CAA AdTdT-3 ' of forward direction
(SEQ ID NO:3), reverse 5 '-UUU GUA GAC GCG GAG CUG GdTdT-3 ' (SEQ ID NO:4)；Target GFP's
SiRNA sequence:5 '-CUU CAG CCU CAG CUU GCC GdTdT-3_ (SEQ ID NO of forward direction:5), reverse 5 '-CGG CAA
GCU GAC CCU GAA GdTdT-3’(SEQ ID NO:6) 2000 transfection reagents of .Lipofectin are purchased from Invitrogen public affairs
Department.
Main method：(1) MDA-MB-231 cells point are in 2 35mm Tissue Culture Dish, so that second day transfection siRNA
Shi Midu reaches 70% or so.Required according to the transfection of Lipofectin 2000, respectively to the MDA- of two 35mm culture dishes
RIPI lysis buffer cell lysis is used after MB-231 cell transfectings siRNA-GFP or siRNA-CD146,48h, does total egg
The white quantitative detection run SDS-PAGE afterwards, and carry out Western Blot progress correlation molecules.Confirm CD146 in MDA-MB-
After the effect that downward is expressed in 231 cells, the tests such as cellular morphology change observation, cell invasion transfer ability are further carried out.
(2) MDA-MB-231 cells point add mIgG and AA98 (50mg/ml) immediately after 2 35mm Tissue Culture Dish
Stimulated, various detections are carried out after 48h, it is and upper same.
We further select triple negative breast cancer cell line MDA-MB-231 (to be purchased from ATCC, numbering HTB-26TM) it is to grind
Study carefully model, the expression of CD146 or anti-CD146 functions are lowered using the siRNA (being synthesized in Invitrogen) of targeting CD146 molecules
Property antibody A A98, it is found that CD146 has critical function to the grade malignancy of tumour cell for maintaining interstitial cell feature.
As a result as shown in Figure 6, Figure 7, the siRNA of CD146 molecules is targeted by CD146 protein levels in MDA-MB-231 cells
It is lowered to original 1/3rd.It is observed that the cell beginning Hui Yuan of shuttle.At the same time, the mark of interstitial cell
Molecule vimentin and Fibronectin expression are lowered, the mark molecule E-cadherin up-regulated expressions of epithelial cell.It is more important
, cell migration and invasive ability are inhibited.Meanwhile anti-CD146 functional antibodies AA98 can also be significantly inhibited
MDA-MB-231 cytoplasm cell marker molecules Slug is expressed, the expression of up-regulation epithelial cell marker molecule E-cadherin,
And suppress cell migration ability.
Sequence table
<110>Institute of Biophysics, Academia Sinica
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cagctggtca acgtggccat ttttggcccc ccttggatgg cattcaagga gaggaaggtg 1320
tgggtgaaag agaatatggt gttgaatctg tcttgtgaag cgtcagggca cccccggccc 1380
accatctcct ggaacgtcaa cggcacggca agtgaacaag accaagatcc acagcgagtc 1440
ctgagcaccc tgaatgtcct cgtgaccccg gagctgttgg agacaggtgt tgaatgcacg 1500
gcctccaacg acctgggcaa aaacaccagc atcctcttcc tggagctggt caatttaacc 1560
accctcacac cagactccaa cacaaccact ggcctcagca cttccactgc cagtcctcat 1620
accagagcca acagcacctc cacagagaga aagctgccgg agccggagag ccggggcgtg 1680
gtcatcgtgg ctgtgattgt gtgcatcctg gtcctggcgg tgctgggcgc tgtcctctat 1740
ttcctctata agaagggcaa gctgccgtgc aggcgctcag ggaagcagga gatcacgctg 1800
cccccgtctc gtaagagcga acttgtagtt gaagttaagt cagataagct cccagaagag 1860
atgggcctcc tgcagggcag cagcggtgac aagagggctc cgggagacca gggagagaaa 1920
tacatcgatc tgaggcatta g 1941
<210> 8
<211> 646
<212> PRT
<213>People's ()
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Met Gly Leu Pro Arg Leu Val Cys Ala Phe Leu Leu Ala Ala Cys Cys
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Cys Cys Pro Arg Val Ala Gly Val Pro Gly Glu Ala Glu Gln Pro Ala
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Pro Glu Leu Val Glu Val Glu Val Gly Ser Thr Ala Leu Leu Lys Cys
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Gly Leu Ser Gln Ser Gln Gly Asn Leu Ser His Val Asp Trp Phe Ser
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Val His Lys Glu Lys Arg Thr Leu Ile Phe Arg Val Arg Gln Gly Gln
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Gly Gln Ser Glu Pro Gly Glu Tyr Glu Gln Arg Leu Ser Leu Gln Asp
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Arg Gly Ala Thr Leu Ala Leu Thr Gln Val Thr Pro Gln Asp Glu Arg
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Ile Phe Leu Cys Gln Gly Lys Arg Pro Arg Ser Gln Glu Tyr Arg Ile
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Gln Leu Arg Val Tyr Lys Ala Pro Glu Glu Pro Asn Ile Gln Val Asn
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Pro Leu Gly Ile Pro Val Asn Ser Lys Glu Pro Glu Glu Val Ala Thr
145 150 155 160
Cys Val Gly Arg Asn Gly Tyr Pro Ile Pro Gln Val Ile Trp Tyr Lys
165 170 175
Asn Gly Arg Pro Leu Lys Glu Glu Lys Asn Arg Val His Ile Gln Ser
180 185 190
Ser Gln Thr Val Glu Ser Ser Gly Leu Tyr Thr Leu Gln Ser Ile Leu
195 200 205
Lys Ala Gln Leu Val Lys Glu Asp Lys Asp Ala Gln Phe Tyr Cys Glu
210 215 220
Leu Asn Tyr Arg Leu Pro Ser Gly Asn His Met Lys Glu Ser Arg Glu
225 230 235 240
Val Thr Val Pro Val Phe Tyr Pro Thr Glu Lys Val Trp Leu Glu Val
245 250 255
Glu Pro Val Gly Met Leu Lys Glu Gly Asp Arg Val Glu Ile Arg Cys
260 265 270
Leu Ala Asp Gly Asn Pro Pro Pro His Phe Ser Ile Ser Lys Gln Asn
275 280 285
Pro Ser Thr Arg Glu Ala Glu Glu Glu Thr Thr Asn Asp Asn Gly Val
290 295 300
Leu Val Leu Glu Pro Ala Arg Lys Glu His Ser Gly Arg Tyr Glu Cys
305 310 315 320
Gln Gly Leu Asp Leu Asp Thr Met Ile Ser Leu Leu Ser Glu Pro Gln
325 330 335
Glu Leu Leu Val Asn Tyr Val Ser Asp Val Arg Val Ser Pro Ala Ala
340 345 350
Pro Glu Arg Gln Glu Gly Ser Ser Leu Thr Leu Thr Cys Glu Ala Glu
355 360 365
Ser Ser Gln Asp Leu Glu Phe Gln Trp Leu Arg Glu Glu Thr Gly Gln
370 375 380
Val Leu Glu Arg Gly Pro Val Leu Gln Leu His Asp Leu Lys Arg Glu
385 390 395 400
Ala Gly Gly Gly Tyr Arg Cys Val Ala Ser Val Pro Ser Ile Pro Gly
405 410 415
Leu Asn Arg Thr Gln Leu Val Asn Val Ala Ile Phe Gly Pro Pro Trp
420 425 430
Met Ala Phe Lys Glu Arg Lys Val Trp Val Lys Glu Asn Met Val Leu
435 440 445
Asn Leu Ser Cys Glu Ala Ser Gly His Pro Arg Pro Thr Ile Ser Trp
450 455 460
Asn Val Asn Gly Thr Ala Ser Glu Gln Asp Gln Asp Pro Gln Arg Val
465 470 475 480
Leu Ser Thr Leu Asn Val Leu Val Thr Pro Glu Leu Leu Glu Thr Gly
485 490 495
Val Glu Cys Thr Ala Ser Asn Asp Leu Gly Lys Asn Thr Ser Ile Leu
500 505 510
Phe Leu Glu Leu Val Asn Leu Thr Thr Leu Thr Pro Asp Ser Asn Thr
515 520 525
Thr Thr Gly Leu Ser Thr Ser Thr Ala Ser Pro His Thr Arg Ala Asn
530 535 540
Ser Thr Ser Thr Glu Arg Lys Leu Pro Glu Pro Glu Ser Arg Gly Val
545 550 555 560
Val Ile Val Ala Val Ile Val Cys Ile Leu Val Leu Ala Val Leu Gly
565 570 575
Ala Val Leu Tyr Phe Leu Tyr Lys Lys Gly Lys Leu Pro Cys Arg Arg
580 585 590
Ser Gly Lys Gln Glu Ile Thr Leu Pro Pro Ser Arg Lys Ser Glu Leu
595 600 605
Val Val Glu Val Lys Ser Asp Lys Leu Pro Glu Glu Met Gly Leu Leu
610 615 620
Gln Gly Ser Ser Gly Asp Lys Arg Ala Pro Gly Asp Gln Gly Glu Lys
625 630 635 640
Tyr Ile Asp Leu Arg His
645
Claims (5)
- The antibody of 1.CD146 or anti-CD146 or the functional form of the antibody are preparing the application in being used to treat the medicine of tumour, Wherein described tumour is metastatic triple negative breast cancer, wherein the antibody of the anti-CD146 is monoclonal antibody AA98.
- 2. application according to claim 1, wherein the functional form of the antibody of the anti-CD146 includes the base of anti-CD146 Because of engineered antibody and humanized antibody.
- 3. according to the application described in any one in claim 1-2, wherein the functional form of the antibody of the anti-CD146 is anti- The Function Coupling thing of the antibody of CD146, wherein with the material of antibody binding be medicine, toxin, radioactivity agent, enzyme, cell because Son or another antibody.
- 4. the antibody of anti-CD146 or the functional form of the antibody are preparing the transfer for suppressing metastatic triple negative breast cancer Application in medicine, wherein the antibody of the anti-CD146 is monoclonal antibody AA98.
- 5. application according to claim 4, wherein the functional form of the antibody of the anti-CD146 includes the base of anti-CD146 Because of engineered antibody and humanized antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711142970.5A CN107929732A (en) | 2011-12-12 | 2011-12-12 | The application of CD146 and its antibody diagnosis and treatment triple negative breast cancer |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711142970.5A CN107929732A (en) | 2011-12-12 | 2011-12-12 | The application of CD146 and its antibody diagnosis and treatment triple negative breast cancer |
| CN201110412778XA CN103157109A (en) | 2011-12-12 | 2011-12-12 | CD146 and antibody diagnosis thereof, and application in treating triple negative breast cancer |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201110412778XA Division CN103157109A (en) | 2011-12-12 | 2011-12-12 | CD146 and antibody diagnosis thereof, and application in treating triple negative breast cancer |
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| CN201711142970.5A Pending CN107929732A (en) | 2011-12-12 | 2011-12-12 | The application of CD146 and its antibody diagnosis and treatment triple negative breast cancer |
| CN201110412778XA Pending CN103157109A (en) | 2011-12-12 | 2011-12-12 | CD146 and antibody diagnosis thereof, and application in treating triple negative breast cancer |
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| CN201110412778XA Pending CN103157109A (en) | 2011-12-12 | 2011-12-12 | CD146 and antibody diagnosis thereof, and application in treating triple negative breast cancer |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113929764A (en) * | 2021-10-15 | 2022-01-14 | 中山大学孙逸仙纪念医院 | Mammary lobular tumor molecular marker CD146 and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105301229B (en) * | 2014-07-22 | 2017-03-15 | 中国医学科学院北京协和医院 | CD146 is as molecular marked compound in diagnosis, the purposes by stages or in prognosis intervention of neuropsychiatric systemic lupus erythematosus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101245101A (en) * | 2008-01-31 | 2008-08-20 | 中国科学院生物物理研究所 | Antihuman CD146 monoclone antibody, composition containing the same, and method for testing dissolubility CD146 |
| WO2009108637A1 (en) * | 2008-02-25 | 2009-09-03 | Prometheus Laboratories, Inc. | Drug selection for breast cancer therapy using antibody-based arrays |
-
2011
- 2011-12-12 CN CN201711142970.5A patent/CN107929732A/en active Pending
- 2011-12-12 CN CN201110412778XA patent/CN103157109A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101245101A (en) * | 2008-01-31 | 2008-08-20 | 中国科学院生物物理研究所 | Antihuman CD146 monoclone antibody, composition containing the same, and method for testing dissolubility CD146 |
| WO2009108637A1 (en) * | 2008-02-25 | 2009-09-03 | Prometheus Laboratories, Inc. | Drug selection for breast cancer therapy using antibody-based arrays |
Non-Patent Citations (1)
| Title |
|---|
| GWLADYS ZABOUO ET AL.: ""CD146 expression is associated with a poor prognosis in human breast tumors and with enhanced motility in breast cancer cell lines"", 《BREAST CANCER RESEARCH》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113929764A (en) * | 2021-10-15 | 2022-01-14 | 中山大学孙逸仙纪念医院 | Mammary lobular tumor molecular marker CD146 and application thereof |
| CN113929764B (en) * | 2021-10-15 | 2023-12-29 | 中山大学孙逸仙纪念医院 | Breast phylliform tumor molecular marker CD146 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103157109A (en) | 2013-06-19 |
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Application publication date: 20180420 |