JP2007074989A - Peptide for inhibiting invasion and metastasis of human cancer cell, polynucleotide for encoding the peptide, and pharmaceutical containing the peptide and polynucleotide - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a remedy for cancer, given by utilizing a molecular target not existed until now, so as to control invasion or metastasis of the cancer, and therefore more efficacious than ever. <P>SOLUTION: A polypeptide which inhibits binding of a proline-rich sequence of AMAP1 to a Src homology domain 3 (SH3) of cortactin is synthesized, for the purpose of inhibiting formation of a protein complex specific in invasive breast cancer and comprising three proteins of AMAP1/paxillin/cortactin. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an atypical SH3 / proline-rich sequence binding inhibition found in a protein-protein interaction specialized for cell invasion, containing a peptide compound or a pharmacologically acceptable salt thereof as an active ingredient. It relates to the agent.

The greatest threat of cancer is its invasion and metastasis. In epithelial cancers, many of the metastases first occur through the invasion of cancer cells into the basement membrane. This has been shown by pathological findings to be prominent in human breast cancer (see Non-Patent Document 1).
In recent years, a number of clinical trials of anticancer drugs targeting the proteolytic enzymes that are known to be involved in the inhibition of invasion and metastasis activity have been carried out at the world level, but their effectiveness has been confirmed. Nothing is equal to nothing. Therefore, researchers around the world are searching for new methods and molecular targets.

The present inventors have found that there is a protein complex specialized in invasive breast cancer that is not detected in human normal mammary epithelium and non-invasive breast cancer (Non-patent Document 2). This complex consists of three proteins, AMAP1 / paxillin / cortactin, whose formation is mediated by Src homology domain 3 (SH3 domain). That is, it binds to the proline-rich sequence of paxillin at the SH3 domain of AMAP1, and binds to the proline-rich sequence of AMAP1 at the SH3 domain of cortactin.
In particular, it has also been found that the binding mode between the cortactin SH3 domain and the proline-rich sequence of AMAP1 is significantly different from other general SH3 / proline binding (Non-patent Document 2).
For example, the proline-rich sequence of cortactin SH3 and dynamin 2 has a 1: 1 binding in the normal binding mode, but the cortactin SH3 domain and the proline-rich sequence of AMAP1 have two molecules of cortactin via the SH3 domain. And one molecule of AMAP1 proline-rich sequence.

  Since SH3 domains are involved in various important aspects including cell growth control and cytoskeleton control, screening of SH3 domain binding inhibitors has been carried out mainly by pharmaceutical companies. However, SH3 domains are present in various proteins, are well conserved structurally, and the binding partner proline peptides are similar to each other. Therefore, even if an inhibitor is screened on the basis of inhibiting one SH3 binding, most of the obtained ones have also inhibited many other SH3 bindings as well. Therefore, even if such an inhibitor is developed as, for example, an anticancer agent, an anti-inflammatory agent, an angiogenesis inhibitor or the like, it is considered that most of them have no strong side effects and are not useful.

Allred DC, Mohsin SK, & Fuqua SA.Endocr.Relat.Cancer, 2001, Mar; 8 (1): 47-61 Onodera Y, Hashimoto S, Hashimoto A, Morishige M, Mazaki Y, Yamada A, Ogawa E, Adachi M, Sakurai T, Manabe T, Wada H, Matsuura N, Sabe H. EMBO J., 2005, Mar 9; 24 ( 5): 963-973 Vives E., Brodin P., & Leblus B. J. Biol. Chem., 1997, Jun 20; 272 (25): 16010-16017 Nagahara H., Vocero-Akbani A. M., Snyder E. L., Ho A., Latham D. G., Lissy N. A., Becker-Hapak M., Ezhevsky S. A., & Dowdy S. F. Nat. Med., Dec; 4 (12): 1449-1452 Vocero-Akbani A., Heyden N. V., Lissy N. A., Ratner L., & Dowdy S. F. Nat. Med., 1999, Jan; 5 (1): 29-33 Schwarze S., Ho A., Vocero-Akbani A., & Dowdy S. F. Science, 1999, Sep 3; 285 (5433): 1569-1572

  An object of the present invention is to provide a means for suppressing cancer invasion / metastasis with respect to cancer treatment. An object of the present invention is to provide a more effective cancer therapeutic agent by utilizing a molecular target that has never existed before.

The present inventors use human breast cancer cells to inhibit the formation of a protein complex consisting of the three AMAP1 / paxillin / cortactin specialized for invasive breast cancer, It was revealed that cell invasion and metastasis can be effectively inhibited.
Therefore, if a polypeptide containing an amino acid sequence corresponding to the fourth sequence of the proline-rich region of AMAP1 (a sequence of 15 amino acid residues from the fifth to the 19th amino acid residues of SEQ ID NO: 1) is used, the proline-rich region of AMAP1 It becomes possible to specifically inhibit the binding between the sequence and the SH3 of the cortactin protein.

  In the present invention, for example, a polypeptide containing the amino acid sequence of SEQ ID NO: 1 is synthesized as a polypeptide containing the amino acid sequence corresponding to the fourth sequence of the proline-rich region of AMAP1, and this peptide is combined with the proline-rich sequence of AMAP1. Competing, it was confirmed that the binding between the proline-rich sequence of AMAP1 and the cortactin protein SH3 was specifically inhibited at a low concentration.

In the present invention, a polypeptide comprising an amino acid sequence obtained by binding a polypeptide sequence derived from the HIV TAT protein of SEQ ID NO: 2 (Non-Patent Documents 3 to 6) to the amino acid sequence of SEQ ID NO: 1 (SEQ ID NO: By using 3), the polypeptide of SEQ ID NO: 1 is devised so that it can be directly introduced into cells simply by addition.
By adding this TAT sequence, (1) the polypeptide can be introduced into the cell simply by adding the polypeptide, (2) the introduction time is short, (3) when administered to an individual, intraperitoneal injection, intravenous injection, etc. There are advantages such as being able to be introduced into the organization. In addition, this TAT sequence can be added to either the N-terminus / C-terminus of the target peptide sequence, etc., and it is free to design a low molecular weight compound based on SEQ ID NO: 1 as a drug in the future. Available to:

  Further, in order to visualize the intracellular localization, fluorescein isothiocyanate (FITC), which is a fluorescent substance, can be added to the C-terminal side of the polypeptide of SEQ ID NO: 3.

The present invention relates to a drug that inhibits the invasion / metastasis activity of human cancer cells, comprising the polypeptide of SEQ ID NO: 1 or SEQ ID NO: 3 as an active ingredient. A composition containing such a polypeptide can be a drug that inhibits the invasion / metastasis activity of human cancer cells by inhibiting the formation of a protein complex specialized in human cancer.
The target cancers are cancers derived from epithelial tissues of any organ that undergoes an infiltration process through the basement membrane, including breast cancer.

These polypeptides may be administered in the form of polypeptides, but DNAs encoding these polypeptides may be transfected into cells by the gene transfer method and expressed in the cells.
Accordingly, the present invention relates to the polynucleotide of SEQ ID NO: 4 that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1.
The present invention also relates to a drug comprising the polynucleotide of SEQ ID NO: 4 as an active ingredient.

In the present invention, it is possible to suppress cancer invasion activity and metastasis activity very effectively by targeting an interface related to protein interaction that has not been tried until now and inhibiting the formation of the protein complex. Indicated. Each identified interface involves a protein interaction module called the SH3 region.
In the present invention, the protein interaction is inhibited by using a peptide. Based on this finding, a low molecular weight compound is designed and synthesized, or the protein interaction is inhibited by binding to these modules. It becomes possible to screen compounds, and in the future, it is expected to be able to promote the development of effective drugs for various epithelial tissue-derived cancers including breast cancer.

(1) Production of polypeptide by chemical synthesis method It is convenient to produce the peptide of the present invention by chemical synthesis method.
As a chemical synthesis method, the Merrifield solid peptide synthesis method is convenient. In this method, first, a t-butoxycarbonyl (Boc) derivative of the first amino acid at the carboxy terminus of the amino acid sequence to be synthesized is introduced into chloromethylated crosslinked polystyrene. Next, a second Boc-amino acid is introduced into the amino group on the resin obtained by removing the Boc group. When this operation is repeated and the target peptide chain is constructed, all protecting groups are removed and the peptide is cleaved from the resin.

(2) Production of polypeptide by gene recombination method It is convenient to produce the polypeptide of the present invention by gene recombination method.
A polynucleotide comprising a polypeptide comprising the amino acid sequence of SEQ ID NO: 3, eg a polynucleotide comprising the amino acid sequence of recombinant SEQ ID NO: 3 in a suitable host system, including a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 4 An expression vector expressing the peptide can be constructed. A host cell is transformed with the obtained expression vector, and this transformant is cultured under conditions suitable for expression of a DNA encoding a polypeptide containing the amino acid sequence of SEQ ID NO: 3. A polypeptide comprising the amino acid sequence of can be produced.

An expression vector containing a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 3 can be constructed by methods known to those skilled in the art. A vector suitable for the expression of a polypeptide comprising the amino acid sequence of SEQ ID NO: 3 will have a promoter for initiation of transcription immediately upstream of the insertion site of the polynucleotide.
Appropriate promoters are also known in the art and can be selected according to functional properties in the host cell. For example, an SV40 virus early gene promoter, a peptide chain elongation factor EF-1α promoter, a metallothionein gene promoter, a β-actin promoter, a CMV virus promoter, etc. can be expressed in an animal cell system. A galactosidase gene promoter or the like can be used for expression in bacteria and Escherichia coli.
It is desirable that there is a transcription termination signal downstream of the insertion site of the polynucleotide encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 3.

  Desirably, a selectable marker such as a drug resistance marker is present in the vector. Or you may transform simultaneously using the plasmid which codes drug resistances, such as an expression vector containing a human hematopoietic-type PGD synthetase, and separate antibiotics.

  To construct an expression vector, a polynucleotide encoding a polypeptide containing the amino acid sequence of SEQ ID NO: 3 is inserted into an appropriate vector. Appropriate vectors are selected from those known in the art in view of promoters, transcription termination signals, selectable markers and other conditions. As a vector that can be used for the purpose of expressing a polynucleotide by inserting a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 3 and introducing it into cultured cells, for example, pKCR in the expression in animal cells , PEF-BOS, CDM8, pCEV4 bovine papilloma virus DNA, and the like in Escherichia coli, pGEMEX, pUC, pTAT and the like can be mentioned.

  A cell that can be used for the expression of a polypeptide comprising the amino acid sequence of SEQ ID NO: 3 is replicable, and is a polynucleotide in which a nucleotide sequence derived from the HIV TAT protein of SEQ ID NO: 5 is added to the nucleotide sequence of SEQ ID NO: 4. Any substance capable of expressing nucleotides may be used. For example, prokaryotic microorganisms such as E. coli; Eukaryotic microorganisms such as cerevisiae and even mammalian cells are used. Tissue culture cells include avian cells or mammalian cells such as mice, rats and monkeys. Selection and use methods of suitable host cell-vector systems are known to those skilled in the art, and any system suitable for expression of a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 2 can be selected from them. Can be selected.

A desired protein can be obtained by culturing the transformed cells according to a conventional method. The medium used for the culture can be appropriately selected according to the properties of the host. For example, when the host is Escherichia coli, the LB medium or TB medium is appropriately selected. When the host is a mammalian cell, RPMI 1640 medium or the like is appropriately selected. Can be used.
Isolation and purification of the protein used in the present invention from the culture obtained by this culture can be performed by conventional methods. For example, the culture is subjected to various treatment operations utilizing physical and chemical properties of the protein. Can be used. Specifically, treatment with a protein precipitating agent, ultrafiltration, high performance liquid chromatography, centrifugation, electrophoresis, affinity chromatography and the like can be used alone or in combination.

(3) Method for Introducing the Polypeptide of SEQ ID NO: 3 into Cultured Cells In the method for introducing a peptide used in the present invention, the type of cell is not particularly limited. However, the introduction efficiency varies depending on the cell type. The efficiency of introduction into cells can be evaluated by the fluorescence intensity of FITC added to the C-terminal side of the polypeptide of SEQ ID NO: 3. The concentration of the introduced substance is in the range of a final concentration of 1 μM to 100 μM. In addition, an introduction time of about 30 minutes to 2 hours is sufficient.

(4) Method for Introducing the Polypeptide of SEQ ID NO: 3 into the Living Body In the same manner as described above, in the method for introducing a peptide used in the present invention, 1-10 mg / kg peptide is required when introducing it into a mouse or rat. And As the administration method, intraperitoneal injection, subcutaneous injection, intravenous injection and the like are possible. Oral administration is also considered. The efficiency of introduction into each tissue can be evaluated by the fluorescence intensity of FITC added to the C-terminal side of the polypeptide of SEQ ID NO: 3.

(5) Preparation of a preparation comprising the polypeptide of SEQ ID NO: 3 as an active ingredient The polypeptide of SEQ ID NO: 3 can be mixed and diluted with a carrier known per se and formulated as a liquid preparation, for example. The liquid preparation can be prepared, for example, using purified water, physiological saline, alcohols such as ethanol / propylene glycol / glycerin / polyethylene glycol, and solvents such as triacetin. The liquid thus prepared can be used by diluting, for example, a lactate Ringer's solution, a maintenance solution composed of an electrolyte solution for infusion, a postoperative recovery solution, a dehydration replenisher, a drip saline solution, or the like. Additives appropriately selected and used as a normal solution, for example, pH adjusting buffer (for example, phosphate buffer, borate buffer, citrate buffer, tartrate buffer, acetate buffer, etc.), isotonic An agent (for example, sorbitol, glycerin, polyethylene glycol, propylene glycol, glucose, sodium chloride, etc.) may be added.
Such formulations further include pharmaceutically acceptable benzalkonium chloride, paraoxybenzoates, benzyl alcohol, parachlormethaxinol, chlorcresol, phenethyl alcohol, sorbic acid or salts thereof, thimerosal, chlorobutanol, etc. Adjuvants such as suitable antiseptic fungicides, wetting agents such as talc, emulsifiers such as polyethylene glycol monooleate, dispersants, stabilizers such as sulfites, hydrogen sulfites and metabisulfites may be added. It can also be said that one of the preferable dosage forms is to administer as a suspending agent using gum arabic, kaolin, methyl cellulose, sodium carboxymethyl cellulose or the like as a suspending agent.

  The administration route is preferably parenteral administration, and includes, for example, intravenous administration, transcerebral spinal fluid administration, transarterial local administration by catheter, or surgical local administration. The dosage of the active ingredient of the present invention is 0.1-1000 mg / kg / day, preferably 1-500 mg / kg / day.

(6) Method for Gene Transfer of Polynucleotide of SEQ ID NO: 4 The present invention relates to a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, which is an agent that inhibits the invasive activity and metastatic activity of human cancer cells The present invention relates to a drug containing the encoding polynucleotide.
Examples of the polynucleotide encoding the peptide containing the amino acid sequence of SEQ ID NO: 1 include a polynucleotide containing the nucleotide sequence shown in SEQ ID NO: 4.
These polynucleotides are introduced into cells by a gene transfer method, and the polypeptides are expressed in the cells. Examples of gene introduction methods include microinjection method, calcium phosphate method, electroporation method, protoplast fusion method, DEAE dextran method, lipofection method, virus method using virus vector, particle gun method and the like. A preferred method is the lipofection method. In this method, a DNA-bound liposome is used to introduce a polynucleotide into a cell using cell fusion. When a cationic liposome made of an artificially produced lipid bilayer and a polynucleotide are mixed, a liposome-polynucleotide complex is formed. When cells are cultured in a solution containing this complex, the polynucleotide in the complex is taken up by the cells. Lipofectin (trade name), lipofectamine (trade name), and the like can be preferably used as artificial lipids for producing cationic liposomes.

Example 1: AMAP1 / cortactin complex formation inhibition test
Confirmation of the formation of AMAP1 / cortactin and AMAP1 / paxillin complex using AMAP1, paxillin and cortactin expressed / purified in E. coli, and AMAP1-SH3 domains, which were forcibly expressed in COS7 cells, and the amino acid of SEQ ID NO: 3 Examination of the inhibitory effect of complex formation by a polypeptide obtained by adding FITC to the C-terminal side of the sequence (hereinafter referred to as “P4-TAT-FITC peptide”, simply abbreviated as “P4” in the figure) Was performed by a pull-down method.

  Glutathione-S-transferase (GST) -tagged cortactin or AMAP1-SH3 domain cDNA is introduced into E. coli, expression is induced by a conventional method, and GST-cortactin or GST is added by glutathione sepharose (Amersham Pharmacia Biotech). -The AMAP1-SH3 domain was purified.

COS7 cells were forced to express by introducing AMAP1, dynamin 2 or paxillin cDNA with a green fluorescein (GFP) tag added using lipofection. Polyfect (manufactured by Qiagen) was used for introduction. 36 hours after gene transfer, the cell extract was treated with 1% NP-40 buffer (1% Nonidet-40, 150 mM NaCl, 20 mM Tris-HCl, pH 7.4, 5 mM EDTA, 1 mM Na ₃ VO ₄ , 1 mM phenylmethyl Prepared with sulfonyl chloride, 5 μg / mL aprotinin, 2 μg / mL leupeptin and 3 μg / mL pepstatin A).

  To 300 μg of the cell extract, P4-TAT-FITC peptide was added, 5 μg of purified GST-cortactin or GST-AMAP1-SH3 domain and 10 μL of glutathione sepharose were added, and incubated at 4 ° C. for 2 hours. The AMAP1 / cortactin complex adsorbed on glutathione sepharose was purified with 1% NP-40 buffer. To each of the purified SH3 domain-binding protein bead complexes, 30 μL of Remli sample buffer [see Protein Chemistry I-Separation and Purification, Tokyo Chemical Dojin, page 211 (1976)] was added and mixed well. Heat at 3 ° C. for 3 minutes. After the beads were precipitated for 3 minutes at 25 ° C. and 1,000 rpm by a rotary centrifuge, the supernatants were each subjected to 8% polyacrylamide gel electrophoresis.

Subsequently, a protein containing a proline-rich sequence in each SH3 domain binding protein complex in the supernatant was detected by Western blot as follows. First, the protein in the gel after electrophoresis was blotted on a 0.45 μm nitrocellulose membrane (Immobilon; manufactured by Millipore), and then TBST containing 0.1% Tween-20, 150 mM NaCl, (10 mM Tris-HCl) was applied to block nonspecific binding.
For detection of a protein containing a proline-rich sequence, mouse anti-GFP antibody, diluted 1: 1000, was reacted for 2 hours. After washing with TBST, an HRP-conjugated anti-mouse antibody (manufactured by Jackson) diluted 1: 10,000 was reacted for 1 hour. After washing with TBST, detection was performed by chemiluminescence using an ECL reagent (Amersham Pharmacia Biotech).
FIG. 1 shows a Western blot electrophoretic diagram showing the formation of AMAP1 / cortactin and dynamin 2 / cortactin complexes and the inhibition of complex formation by P4-TAT-FITC peptide. FIG. 2 shows a Western blot electrophoretic diagram showing the formation of the AMAP1 / paxillin complex and the inhibition of the complex formation by the P4-TAT-FITC peptide.

As a result, as can be seen from FIGS. 1 and 2, it was confirmed that the P4-TAT-FITC peptide specifically inhibits AMAP1 / cortactin complex formation at a low concentration.
According to FIG. 1, it was confirmed that the addition of 10 μM P4-TAT-FITC peptide inhibited the binding between AMAP1 and cortactin, and the addition of 30 μM or more P4-TAT-FITC peptide completely inhibited this binding. It was done.
On the other hand, it was confirmed that the addition of P4-TAT-FITC peptide exceeding 100 μM also inhibited the binding between dynamin 2 and cortactin.
Furthermore, according to FIG. 2, it was confirmed that the addition of P4-TAT-FITC peptide exceeding 100 μM also inhibits the binding between AMAP1 and paxillin.

  From the above results, it was confirmed that the binding between AMAP1 and cortactin is specifically inhibited when the amount of the peptide of the present invention is in the range of 10 to 100 μM, and more preferably in the range of 30 to 100 μM. It was.

Example 2: Invasion activity inhibition test of breast cancer cells The effect of mouse breast cancer cells 4T1 / luc on the invasion activity on Matrigel was examined by Boyden Chamber Assay. P4-TAT-FITC peptide was added to the cells, seeded on a Boyden chamber coated with matrigel, and then the invasive activity at 12 hours was measured. As a control, the amino acid sequence of SEQ ID NO: 2 was added to the C-terminal side of the amino acid sequence obtained by randomly rearranging the amino acid sequence of SEQ ID NO: 1, and FITC was further added to the C-terminal side. Peptide (hereinafter referred to as “scramble-TAT-FITC peptide”. In the figure, simply abbreviated as “scramble”), seeded on a Boyden chamber coated with matrigel, and then measured invasive activity for 12 hours did.
The result is shown in FIG. The vertical axis shows the relative invasion activity of each treated cell when the Matrigel invasion activity of untreated cells is taken as 100.

Simultaneously, MTS assay kit (3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium assay for cytotoxicity of compounds Evaluation was made using a kit (Promega).
The result is shown in FIG. A vertical axis | shaft shows the relative survival rate of each processing cell when the MTS activity which an untreated cell shows is set to 100. FIG.

  As can be seen from FIGS. 3 and 4, in the case of the P4-TAT-FITC peptide, the invasive activity was greatly reduced without affecting the cytotoxicity.

Example 3: Inhibition test on breast cancer cell metastasis in mice (in vivo test)
The influence of mouse 4T1 / luc cells on the lung metastasis activity in Balb / c mice was investigated. 4T1 / luc cells are mouse breast cancer cells in which luciferase is constantly expressed. Metastasis activity is 1 × 10 ⁶ cells injected into the right cervical breast adipose tissue of Balb / c mice (female, 6-8 weeks old). Evaluation was made by measuring the luciferase activity in the liquid (FIG. 5).
11 days after 4T1 / luc cell injection with P4-TAT-FITC peptide at final concentrations of 1, 3 and 10 mg / kg or scrambled-TAT-FITC peptide at final concentration of 10 mg / kg The mice were intraperitoneally administered daily from the eyes.
The weight of the tumor formed at the original injection site on the 19th day was measured (FIG. 6).
Moreover, the body weight of the mouse | mouth from the 11th day after cell injection was measured (FIG. 7).
Ten mice were used for each assay.

As a result, as can be seen from FIG. 5, the P4-TAT-FITC peptide inhibited the metastasis of 4T1 / luc cells to the lung in a concentration-dependent manner.
On the other hand, there was no significant change in the weight of the tumor formed at the original injection site and the body weight of the mice under the conditions (FIGS. 6 and 7).

Western blot electrophoresis showing formation of AMAP1 / cortactin complex and dynamin 2 / cortactin complex by P4-TAT-FITC peptide and inhibition of complex formation. Western blot electrophoretic diagram showing formation of AMAP1 / paxillin complex and inhibition of complex formation by P4-TAT-FITC peptide. Graph showing the inhibition of breast cancer cell invasion activity by P4-TAT-FITC peptide: Invasion activity in Boyden chambers coated with matrigel. Graph showing the inhibition of breast cancer cell survival activity by P4-TAT-FITC peptide: MTS activity of cells. The graph which shows suppression of the metastasis | transition to the lung of the breast cancer by P4-TAT-FITC peptide. Change in tumor weight at the site of breast cancer cell injection. Change in body weight of mice from day 11 after breast cancer cell injection.

Claims (8)

  A polypeptide comprising an amino acid sequence corresponding to the fourth sequence of the proline-rich region of AMAP1.   The polypeptide of claim 1 comprising the amino acid sequence of SEQ ID NO: 1.   The polypeptide of claim 1 comprising the amino acid sequence of SEQ ID NO: 3.   A polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1.   The polynucleotide of Claim 4 comprising the nucleotide sequence of SEQ ID NO: 4.   A polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 3.   A drug that inhibits invasion and metastasis of human cancer cells, comprising the polypeptide according to any one of claims 1 to 3 as an active ingredient. A drug that inhibits invasion and metastasis of human cancer cells, comprising the polynucleotide according to any one of claims 4 to 6 as an active ingredient.

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