CN110357946A - It is a kind of inhibit metastases polypeptide and its application - Google Patents
It is a kind of inhibit metastases polypeptide and its application Download PDFInfo
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- CN110357946A CN110357946A CN201910797124.XA CN201910797124A CN110357946A CN 110357946 A CN110357946 A CN 110357946A CN 201910797124 A CN201910797124 A CN 201910797124A CN 110357946 A CN110357946 A CN 110357946A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
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- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
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Abstract
The present invention relates to biomedicine technical fields, and in particular to a kind of polypeptide for inhibiting metastases and its application, the amino acid sequence of the polypeptide are amino acid sequence shown in SEQ ID NO.1-6.Polypeptide of the present invention is a kind of polypeptide with Chemokine CXCL12 interaction, the affinity of the polypeptide and CXCL12 are higher, and the change of CXCL12 secondary structure can be caused, to block CXCL12 and the combination of its receptor CXCR 4, inhibit the transfer of tumour cell and leukaemia cell etc..
Description
The application is that application No. is 201610907839.2, applying date 2016-10-18, a kind of entitled " inhibition
The divisional application of the polypeptide of metastases and its application ".
Technical field
The present invention relates to biomedicine technical fields, are related to a kind of polypeptide, and in particular to a kind of to inhibit the more of metastases
Peptide and its application, a kind of polypeptide with Chemokine CXCL12 interaction and its application in terms of inhibiting cancer migration.
Background technique
Cancer is to seriously endanger a kind of disease of human health, and cancer is a global serious health problems.
In 2007, it is estimated that the whole world has 7,600,000 people to die of cancer.Such as in Britain, cancer causes 126,000 people dead every year,
The people of a quarter dies of cancer.
Cancer migration is the main reason for causing cancer patient dead.Chemotactic factor (CF) is a kind of single-stranded small protein,
By with the interaction of the g protein coupled receptor of cell surface, cause cytoskeleton to be reset, migration with cell is adhered to and invaded
It is related to attack equal behaviors.Wherein, stroma cell derivative factor SDF-1 (stromal cell-derived factor-1), that is,
Chemokine CXCL12 and its specific receptor CXCR4 are sent out in kinds of tumors and the migration of the organ specificity of leukaemia cell
Wave very important effect.Tumour cell and leukemia cell surface height expression Chemokine receptor CXCR4, and chemotactic factor (CF)
CXCL12 high expression in marrow, lymph node and certain organs.The tumour cell of height expression CXCR4 and leukaemia cell exist
Under the chemotaxis of CXCL12, inverse concentration gradient is migrated to the certain organs, such as lung, liver etc. of chemotactic factor (CF) generating source, shape
At organ specific migration (Balkwill F., Semin Immunol, 2003,15,49-55).Therefore, it is targeted using inhibitor
Block the interaction between CXCL12/CXCR4 that can block sticking for cancer cell and stroma cell, increase tumour cell and
The sensibility of chemotherapeutics prevents the migration and recurrence of cancer.
Since polypeptide is readily synthesized, it is easy to be metabolized and will not bring toxic side effect and serious immune anti-in human body
It answers, therefore inhibiting using polypeptide the migration of tumour cell is a kind of effective and without side-effects method.CN 105434347 A
A kind of cancer therapeutic polypeptide nano micella is disclosed, the polypeptide is able to suppress cancer migration, which targets cell table
Face CXCR4 receptor, and polypeptide provided by the present invention targets Chemokine CXCL12 in this chemotactic axis of CXCL12/CXCR4
This part, can effectively inhibit the migration of tumour cell when peptide concentration is lower.
Summary of the invention
In order to overcome the deficiencies of the prior art, the purpose of the present invention is to provide it is a kind of inhibit metastases polypeptide and its answer
With the peptide inhibitor can be specifically bound with CXCL12, and affinity is higher, and can cause CXCL12 secondary structure
Change, to block CXCL12 and the combination of its receptor CXCR 4, inhibits the migration of tumour cell and leukaemia cell etc..
In order to achieve that object of the invention, the invention adopts the following technical scheme:
In a first aspect, the present invention provides a kind of polypeptides, which is characterized in that the amino acid sequence of the polypeptide is SEQ ID
Amino acid sequence shown in NO.1-6；
The amino acid sequence is as follows:
SEQ ID NO:1:GVSLSYDCGCDFFRSDV；
SEQ ID NO:2:RRRGGSRGDGVSLSYDCGCDFFRSDV；
SEQ ID NO:3:DDDGGSRGDGVSLSYDCGCDFFRSDV；
SEQ ID NO:4:GGRGDLDWIQRYLRDA；
SEQ ID NO:5:RRRGGRGDLDWIQRYLRDA；
SEQ ID NO:6:DDDGGRGDLDWIQRYLRDA.
The present invention has synthesized a kind of specific recognition CXCL12 for the structure of CXCL12 and the sequence of critical segment, design
Polypeptide, which acts on after CXCL12 the change that can cause CXCL12 secondary structure, therefore the polypeptide can be used as
Allosteric modulators are integrated on the allosteric modulating sites of CXCL12, cause the change of CXCL12 conformation, thus influence CXCL12 with
The conformation in the positive structure site CXCR4 (orthosteric site) blocks the combination of CXCL12 and its receptor CXCR 4, inhibits tumour
The migration of cell and leukaemia cell.
Second aspect, the present invention provides a kind of DNA fragmentations, and it includes the nucleotides sequences of polypeptide described in coding first aspect
Column.
The third aspect, the present invention provides a kind of recombinant vector, the recombinant vector contain at least one copy such as the
DNA fragmentation described in two aspects.
Fourth aspect, the present invention provides a kind of recombinant cell, the recombinant cell contains expression described in the third aspect
Carrier.
5th aspect, the present invention provides polypeptides as described in relation to the first aspect, the nucleotide sequence as described in second aspect,
Recombinant vector as described in the third aspect or the recombinant cell as described in fourth aspect are in the application for inhibiting tumor cell migration.
Preferably, the tumor cell migration includes the lateral transfer of cell and the vertical migration of cell.
Preferably, the tumour cell is that CXCR4 high expresses associated cancer cell.
Preferably, the tumour cell is breast cancer cell and/or leukaemia cell.
Polypeptide of the present invention inhibits the lateral transfer of breast cancer cell, and the longitudinal direction of breast cancer cell and leukaemia cell is inhibited to move
It moves.
6th aspect, the present invention provides a kind of composition, the composition include such as polypeptide as described in relation to the first aspect,
Nucleotide sequence as described in second aspect, the recombinant vector as described in the third aspect or the recombination as described in fourth aspect are thin
Born of the same parents.
7th aspect, the present invention provides the compositions as described in terms of the 6th in preparing treating cancer related drugs
Using.
Preferably, the cancer is that CXCR4 high expresses associated cancer.
Preferably, the tumour cell is breast cancer and/or leukaemia.
In the specific embodiment of the present invention, detected using surface plasmon resonance (SPR) method
The affinity of polypeptide and CXCL12 of the present invention.
In the specific embodiment of the present invention, carry out detecting CXCL12 of the present invention using circular dichroism spectra (CD) method
Secondary structure variation.
In the specific embodiment of the present invention, detection tumour lateral transfer is carried out using cell scratch method.
In the specific embodiment of the present invention, detection tumour vertical migration is carried out using transwell method.
Compared with prior art, the invention has the following advantages:
(1) polypeptide of the present invention is high to the affinity of CXCL12, can be specifically binding on CXCL12, and
Cause the change of CXCL12 secondary structure, to block CXCL12 and the combination of its receptor CXCR 4, inhibits the migration of cancer；
(2) polypeptide of the present invention has as treating or the potentiality of the drug of auxiliary therapy cancer, especially as being directed to
The treatment of both diseases of breast cancer and leukaemia and auxiliary therapy drug.
Detailed description of the invention
Fig. 1 is the inhibiting effect that polypeptide SEQ ID NO:4 (W4) of the present invention induces CXCL12 the lateral transfer of MCF-7 cell
Result figure；
Fig. 2 is the suppression that polypeptide SEQ ID NO:4 (W4) of the present invention induces CXCL12 the lateral transfer of MDA-MB-231 cell
Result figure is used in production；
Fig. 3 is the inhibiting effect that polypeptide SEQ ID NO:4 (W4) of the present invention induces CXCL12 MCF-7 cell vertical migration
Result figure；
Fig. 4 is the suppression that polypeptide SEQ ID NO:4 (W4) of the present invention induces CXCL12 MDA-MB-231 cell vertical migration
Result figure is used in production；
Fig. 5 is inhibition of the polypeptide SEQ ID NO:1-6 (W1-6) of the present invention to CXCL12 HL-60 cells cell vertical migration
Exercising result figure；
Fig. 6 is the inhibition that polypeptide SEQ ID NO:1-6 (W1-6) of the present invention induces CXCL12 U937 cell vertical migration
Exercising result figure；
Fig. 7 is polypeptide SEQ ID NO:4 (W4) of the present invention in conjunction with CXCL12 and its to CXCL12 and MCF-7 cell combination
Inhibiting effect result figure, wherein 7 (a) figures indicate that CXCL12 is integrated to the amount of MCF-7 cell surface, and 7 (b) figures indicate
After CXCL12 and W4 interaction, the amount for being integrated to MCF-7 cell surface is substantially reduced；7 (c) figures indicate W4 in MCF-7 cell
Surface is almost without combination；
Fig. 8 is that polypeptide SEQ ID NO:4 (W4) of the present invention is thin with MDA-MB-231 in conjunction with CXCL12 and its to CXCL12
The inhibiting effect result figure that born of the same parents combine, wherein 8 (a) figures indicate that CXCL12 is integrated to the amount of MDA-MB-231 cell surface, 8 (b)
After figure indicates CXCL12 and W4 interaction, the amount for being integrated to MDA-MB-231 cell surface is substantially reduced；8 (c) figures indicate W4
In MDA-MB-231 cell surface almost without combination.
Specific embodiment
The technical scheme of the invention is further explained by means of specific implementation.Those skilled in the art should be bright
, the described embodiments are merely helpful in understanding the present invention, should not be regarded as a specific limitation of the invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Unless specifically stated otherwise, breast cancer cell line MCF-7, MDA-MB-231 and leukaemia used in following embodiment are thin
Born of the same parents system HL-60, U937 are purchased from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences's cell resource center.
Unless specifically stated otherwise, the solvent of aqueous solution used is sterile ultrapure water in following embodiment.
Unless specifically stated otherwise, phosphate buffer used in following embodiment (PBS) is 1 × PBS solution.
Embodiment 1: the synthesis of polypeptide
According to following table sequent synthesis polypeptide (by Shanghai, Ke Tai Biotechnology Co., Ltd is synthesized, purity 98%), experiment
Before be configured to the mother liquor of suitable concentration.
The polypeptide that table 1 synthesizes
| Title | Sequence | Number |
| W1 | GVSLSYDCGCDFFRSDV | SEQ ID NO:1 |
| W2 | RRRGGSRGDGVSLSYDCGCDFFRSDV | SEQ ID NO:2 |
| W3 | DDDGGSRGDGVSLSYDCGCDFFRSDV | SEQ ID NO:3 |
| W4 | GGRGDLDWIQRYLRDA | SEQ ID NO:4 |
| W5 | RRRGGRGDLDWIQRYLRDA | SEQ ID NO:5 |
| W6 | DDDGGRGDLDWIQRYLRDA | SEQ ID NO:6 |
Embodiment 2: the affinity of polypeptide and CXCL12 experiment
The antibody (RayBiotech, the U.S.) of six polypeptides of W1-W6 and anti-CXCL12 are configured to the water-soluble of 1mg/mL
Liquid takes 5 μ L solution drop on the SPR chip (Plexera, the U.S.) of carboxylated, by polypeptide and is resisted by amino acid condensation reaction
Body is fixed on SPR chip, as stationary phase.It then is respectively the CXCL12 (R&D of 50nM, 100nM and 200nM by concentration
Systems, the U.S.) PBS solution carry out SPR analysis, and calculated equilibrium dissociation constant, as a result such as following table institute as mobile phase
Show.
The affinity of 2 polypeptide of table and CXCL12 antibody to CXCL12
Polypeptide W1-W6 and CXCL12 have stronger affinity as can be seen from Table 2, and and CXCL12 antibody combination
Constant is close.
Influence of the embodiment 3:W4 polypeptide to CXCL12 secondary structure
The aqueous solution for configuring 5 μM of the CXCL12 albumen containing various concentration W4 polypeptide, moves to 0.1cm for solution later
In the ultra-thin quartz colorimetric utensil of light path, CD (J-1500, JASCO, Japan) spectrum of solution, scanning are scanned in 190-260nm
Speed is 200nm/min, bandwidth 2nm, and wavelength interval is 0.5nm.Same sample measurement is averaged three times, and is detained
Except the measuring signal of W4 polypeptide itself.Secondary structure is analyzed using JASCO software later.
Influence of the 3 W4 polypeptide of table to CXCL12 secondary structure
As shown in table 3, with the addition of W4 polypeptide, the helical structure of CXCL12 is gradually converted into folding and random coil,
Illustrate that the interaction of W4 and CXCL12 can cause the change of CXCL12 secondary structure.
Inhibiting effect of the embodiment 4:W4 polypeptide to CXCL12 induction MCF-7 cell lateral transfer
In six orifice plates, every hole contains the RPMI-1640 culture medium of 10% fetal calf serum and 1% mycillin using 2mL
(Gibco, the U.S.) culture 5 × 105A MCF-7 cell, by six orifice plates in 37 DEG C, 5%CO2It is cultivated in the incubator of condition for 24 hours,
MCF-7 cell is grown to when closing on 90% fusion, is then used in the hole of culture plate along straight line scratch with the 10 sterile pipette tips of μ L
For PBS solution by soft washing 3 times of cell, the CXCL12 containing 100ng/mL and various concentration W4 polypeptide is added in rear every hole
Culture medium continues to cultivate.0h after scratch, it is moved for 24 hours with the microscope of 10 × camera lens (IX71, Olympus, Japan) observation cell
The emotionally variation of condition and scratch width, calculates cell migration rate, and the cell migration width that CXCL12 group is only added is set as
100%.As shown in Figure 1, W4 polypeptide, which is added, can effectively inhibit the lateral transfer that MCF-7 cell is induced by CXCL12.
Inhibiting effect of the embodiment 5:W4 polypeptide to CXCL12 induction MDA-MB-231 cell lateral transfer
In six orifice plates, every hole using 2mL containing 10% fetal calf serum and 1% mycillin DMEM culture medium (Gibco,
The U.S.) culture 3 × 105A MDA-MB-231 cell, by six orifice plates in 37 DEG C, 5%CO2It is cultivated in the incubator of condition for 24 hours, MDA-
MB-231 cell is grown to when closing on 90% fusion, is then used in the hole of culture plate along straight line scratch with the 10 sterile pipette tips of μ L
For PBS solution by soft washing 3 times of cell, the CXCL12 containing 100ng/mL and various concentration W4 polypeptide is added in rear every hole
Culture medium continues to cultivate.0h after scratch, for 24 hours with the micro- sem observation cell situation of movement of 10 × camera lens and scratch width
Variation, calculates cell migration rate, and the cell migration width that CXCL12 group is only added is set as 100%.As shown in Fig. 2, it is more that W4 is added
Peptide can effectively inhibit the lateral transfer that MDA-MB-231 cell is induced by CXCL12.
Inhibiting effect of the embodiment 6:W4 polypeptide to CXCL12 induction MCF-7 cell vertical migration
200 μ L are contained 1 × 10 by the MCF-7 cell for harvesting logarithmic growth phase5The culture medium of a cell is added
The upper chamber of the cell transwell (the PET miillpore filter that diameter is 8 μm, Mi Libo, Switzerland), lower room are added 800 μ L and contain
The CXCL12 of 100ng/mL and the culture medium of various concentration W4 polypeptide.24 well culture plates are in 37 DEG C, 5%CO2The incubator of condition
The cell transwell is taken out in middle culture afterwards for 24 hours, and the non-migrating cell in filter membrane upper chamber face is carefully wiped with cotton swab, molten with crystal violet
The fixed cell for moving to cell film lower surface of liquid simultaneously dyes 20min, and with 10 × microscope, (DMI3000B is come after clear water rinses
Card, Germany) observe the cell migrated.Each cell film takes 5 visuals field according to the orientation in up and down, counts in each visual field
The cell number N of migration, calculates cell migration rate after being averaged, the cell migration number that CXCL12 group is only added is set as 100%.Such as
Shown in Fig. 3, W4 polypeptide, which is added, can effectively inhibit the vertical migration that MCF-7 cell is induced by CXCL12.
Inhibiting effect of the embodiment 7:W4 polypeptide to CXCL12 induction MDA-MB-231 cell vertical migration
200 μ L are contained 1 × 10 by the MDA-MB-231 cell for harvesting logarithmic growth phase5The culture medium of a cell is added
The upper chamber of the cell transwell (the PET miillpore filter that diameter is 8 μm), the CXCL12 that 800 μ L contain 100ng/mL is added in lower room
And the culture medium of various concentration W4 polypeptide.24 well culture plates are in 37 DEG C, 5%CO2It cultivates in the incubator of condition and takes out afterwards for 24 hours
The cell transwell carefully wipes the non-migrating cell in filter membrane upper chamber face with cotton swab, is fixed with crystal violet solution and move to cell
The cell of film lower surface simultaneously dyes 20min, clear water rinse after with 10 × cell that migrates of micro- sem observation.Each cell film according to
Orientation in up and down takes 5 visuals field, counts the cell number N migrated in each visual field, calculates cell migration after being averaged
Rate, the cell migration number that CXCL12 is only added are set as 100%.As shown in figure 4, W4 polypeptide, which is added, can effectively inhibit MDA-
The vertical migration that MB-231 cell is induced by CXCL12.
Inhibiting effect of the embodiment 8:W1-W6 polypeptide to CXCL12 HL-60 cells cell vertical migration
200 μ L are contained 2 × 10 by the HL-60 cell for harvesting logarithmic growth phase5The culture medium of a cell is added
The upper chamber of the cell transwell (the PET miillpore filter that diameter is 8 μm), the CXCL12 that 800 μ L contain 200ng/mL is added in lower room
And the culture medium of various concentration W4 polypeptide.24 well culture plates are in 37 DEG C, 5%CO2It cultivates in the incubator of condition and takes out afterwards for 24 hours
The cell transwell, counts the cell number N of each lower room migration, and calculates cell migration rate, and CXCL12 group is only added
Cell migration number is set as 100%.As shown in figure 5, W4 polypeptide, which is added, can effectively inhibit what HL-60 cell was induced by CXCL12
Vertical migration.
Inhibiting effect of the embodiment 9:W1-W6 polypeptide to CXCL12 induction U937 cell vertical migration
200 μ L are contained 2 × 10 by the U937 cell for harvesting logarithmic growth phase5Transwell is added in the culture medium of a cell
The upper chamber of cell (the PET miillpore filter that diameter is 5 μm), the CXCL12 and difference that 800 μ L contain 200ng/mL is added in lower room
The culture medium of concentration W4 polypeptide.24 well culture plates are in 37 DEG C, 5%CO2It is cultivated in the incubator of condition and takes out transwell afterwards for 24 hours
Cell, counts the cell number N of each lower room migration, and calculates cell migration rate, and the cell migration of CXCL12 group is only added
Number is set as 100%.As shown in fig. 6, W4 polypeptide, which is added, can effectively inhibit the vertical migration that U937 cell is induced by CXCL12.
Embodiment 10:W4 polypeptide is in conjunction with CXCL12 to the depression effect of CXCL12 and MCF-7 cell combination
1mL is contained 1 × 105The culture medium of a MCF-7 cell is added in Glass bottom culture dish, overnight after cell is adherent,
Will containing 1 μM of Rhodamine B isothiocyanate (RBITC, Beijing Abace Biotechnology Co., Ltd.) label CXCL12,1 μM it is different
The W4 and contain 1 μM of RBITC- simultaneously that thiocyanic acid fluorescein (FITC, Shanghai Ke Tai Biotechnology Co., Ltd) marks
The culture medium of CXCL12 and 1 μM of FITC-W4 is separately added into culture dish, after 37 DEG C of effect 1h with PBS wash away unbonded albumen and
Polypeptide is added nucleus dyestuff Hoechst.33342 (Sigma-Aldrich, the U.S.) and dyes 10min, burnt aobvious in 40 × copolymerization
It takes pictures under micro mirror (LSM760, Zeiss, Germany).As shown in Fig. 7 (a) and 7 (b), after W4 is added, it is integrated to MCF-7 cell table
The amount of the RBITC-CXCL12 in face significantly reduces, and Fig. 7 (c) shows FITC-W4 in MCF-7 cell surface almost without knot
Close, this illustrate W4 polypeptide be by interacting to inhibit the migration of MCF-7 cell with CXCL12, rather than with cell table
The effect that face CXCR4 is combined.
Embodiment 11:W4 polypeptide is in conjunction with CXCL12 to the depression effect of CXCL12 and MDA-MB-231 cell combination
1mL is contained 1 × 105The culture medium of a MDA-MB-231 cell is added in Glass bottom culture dish, pastes overnight to cell
After wall, 1 μM RBITC-CXCL12 and 1 μM will be contained containing 1 μM of RBITC-CXCL12,1 μM of FITC-W4 and simultaneously
The culture medium of FITC-W4 is separately added into culture dish, washes away unbonded albumen and polypeptide after 37 DEG C of effect 1h with PBS, is added thin
Karyon dyestuff Hoechst.33342 dyes 10min, takes pictures under 40 × Laser Scanning Confocal Microscope.As shown in Fig. 8 (a) and 8 (b), W4
After addition, the amount for being integrated to the RBITC-CXCL12 of MDA-MB-231 cell surface is significantly reduced, and Fig. 8 (c) is shown
FITC-W4 in MDA-MB-231 cell surface almost without combination, this illustrate W4 polypeptide be by with CXCL12 interaction from
And the migration of MDA-MB-231 cell is inhibited, rather than the effect in conjunction with cell surface CXCR4.
Above-mentioned experiment is shown: polypeptide of the present invention can be specifically bound with CXCL12, and affinity is higher, and should
Polypeptide can cause the change of CXCL12 secondary structure after acting on CXCL12, to block CXCL12 and its receptor CXCR 4
Combination, inhibit the migration of cancer.The polypeptide provides feasible treatment method for the inhibition that cancer migrates.
The Applicant declares that the present invention illustrates the process method of the present invention through the above embodiments, but the present invention not office
It is limited to above-mentioned processing step, that is, does not mean that the present invention must rely on the above process steps to be carried out.Technical field
Technical staff it will be clearly understood that any improvement in the present invention, equivalence replacement and auxiliary element to raw material selected by the present invention
Addition, selection of concrete mode etc., all of which fall within the scope of protection and disclosure of the present invention.
SEQUENCE LISTING
<110>State Nanometer Science Center
<120>a kind of polypeptide for inhibiting metastases and its application
<130> 2016
<160> 6
<170> PatentIn version 3.3
<210> 1
<211> 17
<212> PRT
<213>artificial synthesized sequence
<400> 1
Gly Val Ser Leu Ser Tyr Asp Cys Gly Cys Asp Phe Phe Arg Ser Asp
1 5 10 15
Val
<210> 2
<211> 26
<212> PRT
<213>artificial synthesized sequence
<400> 2
Arg Arg Arg Gly Gly Ser Arg Gly Asp Gly Val Ser Leu Ser Tyr Asp
1 5 10 15
Cys Gly Cys Asp Phe Phe Arg Ser Asp Val
20 25
<210> 3
<211> 26
<212> PRT
<213>artificial synthesized sequence
<400> 3
Asp Asp Asp Gly Gly Ser Arg Gly Asp Gly Val Ser Leu Ser Tyr Asp
1 5 10 15
Cys Gly Cys Asp Phe Phe Arg Ser Asp Val
20 25
<210> 4
<211> 16
<212> PRT
<213>artificial synthesized sequence
<400> 4
Gly Gly Arg Gly Asp Leu Asp Trp Ile Gln Arg Tyr Leu Arg Asp Ala
1 5 10 15
<210> 5
<211> 19
<212> PRT
<213>artificial synthesized sequence
<400> 5
Arg Arg Arg Gly Gly Arg Gly Asp Leu Asp Trp Ile Gln Arg Tyr Leu
1 5 10 15
Arg Asp Ala
<210> 6
<211> 19
<212> PRT
<213>artificial synthesized sequence
<400> 6
Asp Asp Asp Gly Gly Arg Gly Asp Leu Asp Trp Ile Gln Arg Tyr Leu
1 5 10 15
Arg Asp Ala
Claims (10)
1. a kind of polypeptide, which is characterized in that the amino acid sequence of the polypeptide is amino acid shown in SEQ ID NO.1-3,5-6
Sequence.
2. a kind of DNA fragmentation, which is characterized in that it includes the nucleotide sequences of polypeptide described in coding claim 1.
3. a kind of recombinant vector, which is characterized in that the recombinant vector contains the as claimed in claim 2 of at least one copy
DNA fragmentation.
4. a kind of recombinant cell, which is characterized in that the recombinant cell contains expression vector as claimed in claim 3.
5. polypeptide as described in claim 1, nucleotide sequence as claimed in claim 2 is as claimed in claim 3 to recombinate
Carrier or recombinant cell as claimed in claim 4 are in the application for inhibiting tumor cell migration.
6. application according to claim 5, which is characterized in that the tumor cell migration include cell lateral transfer and
The vertical migration of cell.
7. application according to claim 5, which is characterized in that the tumour cell is that CXCR4 high expression associated cancer is thin
Born of the same parents；
Preferably, the tumour cell is breast cancer cell and/or leukaemia cell.
8. a kind of composition, which is characterized in that the composition includes polypeptide as described in claim 1, such as claim 2 institute
The nucleotide sequence stated, recombinant vector as claimed in claim 3 or recombinant cell as claimed in claim 4.
9. application of the composition according to claim 8 in preparation treating cancer related drugs.
10. application according to claim 9, which is characterized in that the cancer is that CXCR4 high expresses associated cancer；
Preferably, the tumour cell is breast cancer and/or leukaemia.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910797124.XA CN110357946B (en) | 2016-10-18 | 2016-10-18 | Polypeptide for inhibiting tumor metastasis and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910797124.XA CN110357946B (en) | 2016-10-18 | 2016-10-18 | Polypeptide for inhibiting tumor metastasis and application thereof |
| CN201610907839.2A CN106432424B (en) | 2016-10-18 | 2016-10-18 | It is a kind of inhibit metastases polypeptide and its application |
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| CN201610907839.2A Division CN106432424B (en) | 2016-10-18 | 2016-10-18 | It is a kind of inhibit metastases polypeptide and its application |
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| Publication Number | Publication Date |
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| CN110357946A true CN110357946A (en) | 2019-10-22 |
| CN110357946B CN110357946B (en) | 2021-08-03 |
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| CN201910797124.XA Active CN110357946B (en) | 2016-10-18 | 2016-10-18 | Polypeptide for inhibiting tumor metastasis and application thereof |
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| CN106432424A (en) | 2017-02-22 |
| CN106432424B (en) | 2019-11-12 |
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