KR20150142122A - Use of HOXB5 for diagnosis and treatment of breast cancer - Google Patents

Abstract

The present invention relates to a composition comprising a HOXB5 gene inhibitor as an active ingredient for treating breast cancer, or to a composition for diagnosing by using the same. In the case of using the HOXB5 of the present invention as a marker for diagnosing breast cancer, the composition can be significantly used for early detection of breast cancer, effective treatment, and diagnosis, by diagnosing estrogen receptor-positive breast cancer which comprises 50-80% of the entire patient with breast cancer.

Description

Use of HOXB5 for diagnosis and treatment of breast cancer [

To a composition for treating breast cancer, or a diagnostic composition using the same.

Breast cancer is the most common cancer in women all over the world, and is the fifth most common form of cancer among men and women. In breast cancer, early detection is more important than other cancers, since once the cancer cells invade the surrounding tissues or metastasize to the lymph nodes, curing is difficult.

In order to reduce the mortality rate from breast cancer, it is important to first diagnose breast cancer early, and secondly to diagnose the prognosis after primary surgery and to perform appropriate adjuvant therapy. In addition to self-diagnosis by first-order promotion, mammography, mammography and ultrasound are used as preventive measures for breast cancer diagnosis, and this method is the most widely used method for early breast cancer diagnosis . However, mammography has a disadvantage in that the diagnosis rate is low due to the high amount of fiber in the case of dense breast, which is commonly found in Korean women. In addition, because of the use of X-rays, it is not possible to rule out the possibility of breast cancer in the diagnosis process. So ultrasound is being used as an alternative, but it is also difficult to distinguish between malignant and non-cancer. In case of abnormal clinical findings, fine needle aspiration cytology, magnetic resonance imaging (MRI) are additionally used to improve diagnosis rate. However, even using these methods, it is difficult to distinguish malignant tumors from non-cancer malignant tumors, because normal and non-normal tissues are classified as morphologically. For these reasons, breast cancer has developed a method of diagnosing breast cancer in a relatively molecular genetic way as compared to other cancers.

However, amplification and overexpression analysis of HER2 / neu (also known as Human Epidermal Growth Factor Receptor 2, ErbB-2 or ERBB2) gene, which is the most useful breast cancer diagnostic marker for diagnosis and treatment of breast cancer, Only 35% is useful. Estrogen receptor positive (ER +) breast cancer, which accounts for 50% to 80% of breast cancer, has not progressed in the development of genetic markers in its diagnosis and progression.

Breast cancer has a different treatment method depending on the presence or absence of estrogen receptors and HER2 / neu gene. Therapeutic methods for estrogen receptor and HER2 / neu gene are different.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art, and an object of the present invention is to provide a composition for diagnosing breast cancer that is positive for estrogen receptor type, comprising the agent for measuring the expression level of mRNA of HOXB5 (hemeobox B5) It is an object of the present invention to provide an estrogen receptor-positive breast cancer diagnostic kit comprising the composition, and to provide an estrogen receptor positive breast cancer diagnosis method and screening method using the same.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

The term " HOX " in the present invention is a gene coding for a family of transcription regulatory elements called homeobox genes. In vertebrates, there are four groups on different chromosomes, HOX A, B, C and D, (Krumlauf R. Hox genes in vertebrate development. Cell 1994. 78: 191-201). Each HOX gene is classified into 13 groups according to the similarity of the amino acid sequence of the homeomegal region and is described as HOXA1, HOXD13, and the like. The HOX gene is a gene that works in the development of plants and animals. When a mutation occurs, it causes abnormal morphogenesis. The dysregulated expression of HOX genes is found in a tissue-specific manner in cancer. For example, in the case of the HOXB family, HOXB2 is associated with pancreatic cancer and breast cancer, HOXB3 is acute myelogenous leukemia, HOXB5 acute myelogenous leukemia and congenital cystic adenomatoid malformation (CCAM), HOXB6 is acute myelogenous leukemia and colon cancer, HOXB7 HOXB8 is associated with colon cancer, HOXB9 is associated with prostate cancer and lung cancer, and HOXB13 is associated with prostate cancer (http://www.ncbi.nlm.nih.gov/gene). In addition, the dysregulation of HOX genes, which cause morphogenesis abnormalities, can be divided into negative regulation (regulation of gene activation) and positive regulation (promotion of gene activation) according to gene expression method (Shah N , Sukumar S. The Hox genes and their roles in oncogenesis. Nat Rev Cancer 2010; 10: 361-71). For example, HOXB2 is associated with breast cancer due to abnormal voice regulation (Boimel et al., A functional in vivo screen for regulators of tumor progression identifies HOXB2 as a regulator of tumor growth in breast cancer. Genomics. 2011; 98 (3): 164 -72), and HOXB9 causes lung cancer with an abnormal positive control (Calvo et al., Altered HOX and WNT7A expression in human lung cancer, Proc Natl Acad Sci USA 2000; 97 (23): 12776-81). Therefore, the new function of HOX gene is difficult to predict even in the same family, and it is required to continuously develop HOX gene markers capable of specifically diagnosing various cancers.

In one embodiment of the present invention, there is provided a pharmaceutical composition for inhibiting angiogenesis and metastasis comprising an inhibitor of HOXB5 (Homeobox B5) gene expression or an inhibitor of HOXB5 protein activity as an active ingredient. The pharmaceutical composition is a pharmaceutical composition for inhibiting angiogenesis and metastasis for the purpose of inhibiting growth and metastasis of cancer, wherein the gene expression inhibitor is selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific to the HOXB5 gene , Wherein the activity inhibitor of said protein comprises an antibody or an antigen binding fragment thereof specific for the HOXB5 protein.

"Neovascularization" in the present invention means a cell phenomenon in which vascular endothelial cells are proliferated and reconstituted to form new blood vessels from existing vascular networks. In order for cancer cells to proliferate and grow, new blood vessels are needed to supply oxygen and nutrients, thus inhibiting cancer metastasis through inhibition of neovascularization. In addition, "metastasis" refers to the transfer of cancer cells from the primary organs to other organs. The spread of cancer to other parts of the body is largely divided into the development of cancerous tissue in primary cancer, direct invasion of the surrounding organs, and distant metastasis along the blood vessels or lymph vessels to other organs far away. Metastasis can be regulated by inhibiting the expression of a gene associated with cancer development or inhibiting the protein activity of the gene.

In the present invention, the gene can be inhibited to express antisense oligonucleotides complementary to HOXB5, siRNA, shRNA and microRNA.

The antisense oligonucleotides are short length DNA synthesis strands (or DNA analogs) that are antisense (or complementary) to a particular DNA or RNA target and are used to achieve gene-specific inhibition both in vivo as well as ex vivo. Antisense oligonucleotides have been proposed to inhibit the expression of proteins encoded by DNA or RNA targets by binding to the target and stopping expression at the transcription, translation or splicing step. Antisense oligonucleotides have also been successfully used in animal models of cell culture and disease. Other variations of antisense oligonucleotides to make the oligonucleotides more stable and resistant to degradation are known and understood by those skilled in the art. Antisense oligonucleotides used herein include double stranded or single stranded DNA, double stranded or single stranded RNA, DNA / RNA hybrids, DNA and RNA analogs and oligonucleotides with base, sugar or backbone modifications. Oligonucleotides are modified by methods known in the art to increase stability and increase resistance to nuclease degradation. These modifications include, but are not limited to, modifications of the oligonucleotide backbone known in the art, modifications of the sugar moiety, or modifications of the base.

The siRNA (small interfering RNA) is a nucleic acid molecule capable of mediating RNA interference or gene silencing and can be used as an effective gene knockdown method or gene therapy method because it can inhibit the expression of a target gene . When siRNA molecules are used in the present invention, the sense strand (sequence corresponding to the HOXB5 mRNA sequence) and the antisense strand (sequence complementary to the HOXB5 mRNA sequence) are located on opposite sides to form a double-stranded structure or a self- single-stranded structure with sense and antisense strands. siRNAs are not limited to the complete pairing of double-stranded RNA portions that are paired with RNAs, but rather a mismatch (the corresponding base is not complementary), an expansion / bulge (a base corresponding to one strand And a non-paired portion may be included. The siRNA terminal structure is capable of blunt or cohesive termini as long as it can inhibit the expression of HOXB5 gene by RNA interference (RNAi) effect. The sticky end structure can be a 3'-end protruding structure and a 5'-end protruding structure. The siRNA molecule may have a total length of 15 to 30 bases, preferably 19 to 21 bases, although not limited thereto.

The shRNA (short hairpin RNA) is a single-stranded RNA having a length of 45 to 70 nucleotides. The shRNA (short hairpin RNA) is synthesized by synthesizing oligo DNAs connecting 3-10 base linkers between a sense strand of a target gene siRNA base sequence and a complementary antisense strand After shRNA cloning into a plasmid vector or insertion of shRNA into retroviruses such as lentivirus and adenovirus, a hairpin shRNA with a loop is produced and a dicer in the cell is generated. Gt; siRNA < / RTI > to show the RNAi effect.

The microRNA regulates various biological processes such as development, differentiation, proliferation, preservation and apoptosis. MicroRNAs generally regulate the expression of a gene encoding a target mRNA by destabilizing or mobilizing the target mRNA.

Regulatory sequences (e.g., constitutive promoters, inducible promoters, tissue-specific promoters or combinations thereof) useful in expression constructs / vectors with the antisense oligonucleotides, siRNA, shRNA or microRNAs may also be used in the art And the expression inhibitor of the gene may be any substance that inhibits the expression of the gene in addition to the antisense oligonucleotide, siRNA, shRNA or microRNA.

Also, in the present invention, the protein may be inactivated by a HOXB5-specific antibody or an antigen-binding fragment thereof.

As used herein, the term antibody refers to a specific protein molecule that is indicated for an antigenic site as a term known in the art. For the purpose of the present invention, an antibody refers to an antibody that specifically binds to a protein expressed from the HOXB5 gene, which is a marker of the present invention. Such an antibody is obtained by cloning each gene into an expression vector according to a conventional method, , And can be prepared from the obtained protein by a conventional method. Also included are partial peptides that can be made from the protein, and the partial peptides of the invention include at least 7 amino acids, preferably 9 amino acids, more preferably 12 or more amino acids. The form of the antibody of the present invention is not particularly limited, and any polyclonal antibody, monoclonal antibody or antigen-binding antibody thereof may be included in the antibody of the present invention and include all immunoglobulin antibodies. Furthermore, the antibodies of the present invention include special antibodies such as humanized antibodies. Antibodies used in the detection of cancer diagnostic markers of the invention include functional fragments of antibody molecules as well as complete forms with two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having at least an antigen-binding function, and includes Fab, F (ab ') 2, F (ab') 2 and Fv.

In another embodiment of the present invention, an estrogen receptor positive (ER +) type comprising an agent that measures the level of expression of mRNA (SEQ ID NO: 1) or protein (SEQ ID NO: 2) of the HOXB5 (Homeobox B5) There is provided a composition for diagnosing breast cancer. Wherein the agent for measuring the expression level of the mRNA comprises a primer or a probe that specifically binds to the gene, and the agent for measuring the protein expression level of the gene includes an antibody specific for the protein ≪ / RTI >

The term " diagnosis " in the present invention refers to confirming the presence or characteristic of a pathological condition. For the purpose of the present invention, the diagnosis is to confirm the estrogen receptor positive (ER +) type breast cancer.

The term "diagnostic marker, diagnostic marker or diagnostic marker" in the present invention refers to a substance capable of diagnosing cancer cells by distinguishing cancer cells from normal cells, and includes polypeptides that show an increase or decrease in cancer- And organic biomolecules such as nucleic acids (e.g., mRNA), lipids, glycolipids, glycoproteins or sugars (monosaccharides, disaccharides, oligosaccharides and the like). For the purpose of the present invention, the cancer diagnostic marker of the present invention is characterized in that the mRNA of the HOXB5 gene showing a specifically high level of expression in estrogen receptor positive (ER +) type breast cancer cells and the protein encoded thereby to be.

The above-mentioned "measurement of mRNA expression level " is a process of confirming the presence and expression level of mRNA of cancer marker genes in a biological sample in order to diagnose cancer by measuring the amount of mRNA. (RT-PCR), Competitive RT-PCR, Real-time RT-PCR, RNase protection assay (RPA) and Northern blotting Northern blotting, or DNA chip, but are not limited thereto.

The agent for measuring mRNA expression level of the gene means a molecule which can be used for detecting a marker by confirming the expression level of the HOXB5 gene whose expression is increased in cancer cells, Or a primer or a probe that binds to the target. Based on the polynucleotide sequence of HOXB5, one skilled in the art can design primers or probes that specifically amplify specific regions of these genes.

The term "primer" in the present invention refers to a nucleotide sequence having a short free 3 'hydroxyl group, capable of forming a base pair with a template complementary to the template, ≪ / RTI > The primer can initiate DNA synthesis in the presence of reagents and four different nucleoside triphosphates for polymerization reactions (i.e., DNA polymerase or reverse transcriptase) at appropriate buffer solutions and temperatures. In the present invention, it is possible to diagnose cancer through PCR amplification using a sense and antisense primer of HOXB5 polynucleotide to generate a desired product. The PCR conditions, the lengths of the sense and antisense primers can be modified based on what is known in the art.

The term "probe " means a nucleic acid fragment such as RNA or DNA corresponding to a few nucleotides or hundreds of nucleotides that can specifically bind to mRNA, and the presence or absence of a specific mRNA can be confirmed by labeling The probe may be prepared in the form of an oligonucleotide probe, a single stranded DNA probe, a double stranded DNA probe, an RNA probe, etc. In the present invention, a probe complementary to the HOXB5 polynucleotide Hybridization can be carried out using the hybridization method to diagnose cancer through hybridization. The selection and hybridization conditions of suitable probes can be modified based on those known in the art.

The primers or probes of the present invention can be chemically synthesized using the phosphoramidite solid support method, or other well-known methods. Such nucleic acid sequences may also be modified using many means known in the art. Non-limiting examples of such modifications include, but are not limited to, methylation, "capping ", replacement of natural nucleotides with one or more homologues, and modifications between nucleotides, such as uncharged linkers, such as methylphosphonate, Phosphoamidates, carbamates, etc.) or charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.).

The term " measurement of protein expression level " in the present invention is a process for confirming the presence and expression level of a protein expressed from a cancer marker gene in a biological sample in order to diagnose cancer. Preferably, The amount of protein can be confirmed using an antibody that binds to the protein. Examples of the assay methods include Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis But are not limited to, tissue immuno staining, immunoprecipitation assays, complement fixation assays, fluorescence activated cell sorters (FACS), protein chips, and the like.

The agent for measuring the expression level of the protein refers to a molecule that can be used for detecting a marker by confirming the expression level of the protein expressed from the HOXB5 gene, which is a marker for increasing expression in cancer cells, May comprise an antibody specific for the protein. Based on the amino acid sequence of the protein expressed from the HOXB5 gene, one skilled in the art can design an antibody specific for the protein.

In one embodiment of the present invention, the term "antibody" is as described above, and the monoclonal antibody to the protein may be produced and used through a monoclonal antibody production method as is conventional in the art, Can be used. In addition, a polyclonal antibody recognizing the protein may be used instead of the monoclonal antibody, or it may be produced and used through an antiserum preparation method customary in the art.

In another embodiment of the present invention, there is provided an estrogen receptor positive (ER +) type breast cancer diagnostic kit comprising any one of the above compositions. Wherein the kit is selected from the group consisting of a microarray, a gene amplification kit, or a kit for immunoassay.

The term " kit " in the present invention means a set of a composition and accessories necessary for a specific purpose. For the purpose of the present invention, the kit of the present invention can detect the marker by checking the expression level of HOXB5, which is a cancer diagnostic marker, in the expression level of mRNA or its protein. The kit of the present invention may include one or more other component compositions, solutions, or devices suitable for the assay, as well as antibodies recognizing primers, probes or optionally markers for measuring the level of expression of a cancer diagnostic marker.

In another embodiment of the present invention, a pharmaceutical composition for inhibiting angiogenesis and metastasis comprising an inhibitor of HOXB5 (Homeobox B5) gene expression or an inhibitor of the activity of HOXB5 protein as an active ingredient is used to detect estrogen receptor positive 0.0 > (ER +) < / RTI > type breast cancer. The pharmaceutical composition is a method for providing information on treating an estrogen receptor positive (ER +) type breast cancer using an angiogenesis and metastasis inhibiting pharmaceutical composition aimed at inhibiting the growth and metastasis of cancer (ER +) type breast cancer using a composition wherein the gene expression inhibitor is selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific for the HOXB5 gene. (ER +) type breast cancer using a composition wherein the activity inhibitor of said protein comprises an antibody or an antigen binding fragment thereof specific for the HOXB5 protein, and a method for the treatment of estrogen receptor positive Provide a way to provide information The.

"Administration" in the present invention means introducing the composition of the present invention to a patient by any appropriate method, and the administration route of the composition of the present invention can be administered through any conventional route so long as it can reach the target tissue . But are not limited to, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, intranasal administration, intrapulmonary administration, intrarectal administration, intraperitoneal administration, intraperitoneal administration, Do not. The method of treatment of the present invention may comprise administering the pharmaceutical composition in a pharmaceutically effective amount. In the present invention, the effective amount may be determined depending on the kind of the disease, the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of the formulation and the age, body weight, general health condition, sex and diet, And the fraction of the composition, the duration of the treatment, the drug being co-administered, and the like. In the case of an adult, 0.01 ng / kg-10 mg / kg in the case of siRNA, 0.01 ng in the case of the antisense oligonucleotide against mRNA of the gene when the gene expression inhibitor or the protein inhibitor is administered once or several times a day / kg-10 mg / kg, 0.1 ng / kg-10 mg / kg for the compound and 0.1 ng / kg-10 mg / kg for the monoclonal antibody against the protein.

The composition of the present invention contains 0.1 to 50% by weight of the active ingredient of SEQ ID NO: 1 or 2, based on the total weight of the composition. The compositions of the present invention may further comprise suitable carriers, excipients or diluents according to conventional methods. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories or sterilized injection solutions according to a conventional method have. More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. The solid preparation for oral administration includes tablets, pills, powders, granules, capsules and the like. The solid preparation is prepared by mixing at least one excipient such as starch, calcium carbonate (calcium carbonate, sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

In another embodiment of the present invention, an estrogen receptor positive (ER +) type breast cancer diagnostic composition, comprising an agent that measures the level of expression of the HOXB5 (Homeobox B5) gene mRNA or protein, Lt; RTI ID = 0.0 > (ER +) < / RTI > type of breast cancer. Wherein the agent for measuring the level of expression of the mRNA comprises a primer or a probe that specifically binds to the gene, and provides information on diagnosing an estrogen receptor positive (ER +) type breast cancer using a composition Wherein the measurement of the mRNA expression level is performed by a reverse transcriptase polymerase, a competitive reverse transcriptase polymerase, a real-time reverse transcriptase polymerase, an RNase protection assay, a Northern blotting or a DNA chip, Wherein the agent that measures the level of protein expression of said protein comprises an antibody specific for said protein, said method comprising providing information about diagnosing an estrogen receptor positive (ER +) type breast cancer using said composition, Measurement of expression levels was performed by Western blotting, ELISA, radioimmunoassay, Immunoassay, immunoassay, immobilization assay, complement fixation assay, FACS or protein chip. The present invention also provides a method of immunoassay, including immunoassay, diffusion, Ouchteroni immunodiffusion, rocket immunoelectrophoresis,

In another embodiment of the present invention, an estrogen receptor positive (ER +) type breast cancer diagnostic kit is used to diagnose estrogen receptor positive (ER +) type breast cancer, comprising any one of the above compositions Provide a way to provide information about things. The kit can be used to provide information on diagnosing estrogen receptor positive (ER +) type breast cancer using a kit selected from the group consisting of a microarray, a gene amplification kit or an immunoassay kit .

In another embodiment of the present invention, the level of expression of the HOXB5 (Homeobox B5) gene or the level of expression of the protein encoded by the gene after administration of a candidate agent expected to be able to treat an estrogen receptor positive (ER +) type breast cancer (ER < + >) type breast cancer therapeutic, which comprises measuring the expression levels of estrogen receptor positive (ER +) type cancer cells.

In one embodiment of the present invention, in order to inhibit the expression of the HOXB5 (Homeobox B5) gene, an angiogenesis including at least one selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific for the HOXB5 gene as an active ingredient Or a pharmaceutically acceptable salt thereof. In this embodiment, the angiogenesis inhibition provides a pharmaceutical composition that inhibits breast cancer growth, wherein the breast cancer provides a pharmaceutical composition that is estrogen receptor positive, and the HOXB5 (Homeobox B5) gene is an angiogenic inhibitor of SEQ ID NO: 1 A pharmaceutical composition is provided.

In one embodiment of the present invention, there is provided a pharmaceutical composition for inhibiting angiogenesis comprising an HOXB5 specific antibody as an active ingredient for HOXB5 (Homeobox B5) protein inhibition. In this embodiment, angiogenesis inhibition provides a pharmaceutical composition that inhibits breast cancer growth, wherein the breast cancer provides a pharmaceutical composition that is estrogen receptor positive, and the HOXB5 protein provides the pharmaceutical composition of SEQ ID NO: 2.

In one embodiment of the present invention, in order to inhibit the expression of the HOXB5 (Homeobox B5) gene, a cancer metastasis comprising any one or more selected from the group consisting of antisense oligonucleotides, siRNA, shRNA, and microRNAs specific for the HOXB5 gene Or a pharmaceutically acceptable salt thereof. In this embodiment, the suppression of metastasis provides a pharmaceutical composition that inhibits breast cancer growth, and the breast cancer provides a pharmaceutical composition wherein the estrogen receptor is positive.

In one embodiment of the present invention, there is provided a pharmaceutical composition for inhibiting cancer metastasis comprising an antibody specific for HOXB5 as an active ingredient for HOXB5 (Homeobox B5) protein inhibition. In this embodiment, the cancer metastasis inhibition provides a pharmaceutical composition that inhibits breast cancer growth, and the breast cancer provides a pharmaceutical composition that is positive for estrogen receptors.

In one embodiment of the present invention, there is provided an estrogen receptor positive (ER +) type breast cancer diagnostic composition comprising a primer or a probe that specifically binds to the mRNA of the HOXB5 (Homeobox B5) gene, , The HOXB5 (Homeobox B5) gene provides the composition represented by SEQ ID NO: 1.

In one embodiment of the present invention, there is provided an estrogen receptor positive (ER +) type breast cancer diagnostic composition comprising a HOXB5 specific antibody as an active ingredient for HOXB5 (Homeobox B5) protein inhibition. In this embodiment, the HOXB5 protein provides the composition of SEQ ID NO: 2.

In one embodiment of the present invention, the level of the mRNA of HOXB5 (Homeobox B5) gene of the subject is measured with a primer or a probe specifically binding to the mRNA of the HOXB5 (Homeobox B5) gene to determine the estrogen receptor positive ER +) type breast cancer. In this embodiment, the HOXB5 (Homeobox B5) gene provides a method of providing the information represented by SEQ ID NO: 1.

In one embodiment of the present invention, there is provided a method for providing information about estrogen receptor positive (ER +) type breast cancer diagnosis comprising HOXB5 specific antibody as an active ingredient for HOXB5 (Homeobox B5) protein inhibition . In this embodiment, the HOXB5 protein provides a method of providing the information represented by SEQ ID NO: 2.

In one embodiment of the present invention, when the expression level of the HOXB5 (Homeobox B5) gene or the expression level of the protein encoded by the gene is inhibited after administration of estrogen receptor positive (ER +) type breast cancer candidate substance (ER +) type breast cancer drug that determines a candidate substance to be an estrogen receptor positive (ER +) type breast cancer therapeutic agent.

Hereinafter, the present invention will be described in detail.

Breast cancer has a high incidence and rapid growth rate, so accurate and rapid gene diagnosis is required. However, more than 80% of breast cancer patients have estrogen receptor positive (ER +) type breast cancer, Only 30% of HER2 overexpressed genes have been developed. However, the present invention has variously confirmed the association of estrogen receptor positive (ER +) type breast cancer tissues and breast cancer cell lines with significantly higher expression of HOXB5 mRNA and protein. Therefore, the method of diagnosing estrogen receptor positive (ER +) type breast cancer using the HOXB5 gene expression of the present invention can increase the accuracy of diagnosis and can embrace a wide range of whole breast cancer, Estrogen receptor positive (ER +) type breast cancer is expected to be used for diagnosis.

FIG. 1 is a graph showing the results of RTX-PCR for the expression level of HOXB5 in 9 kinds of breast cancer cell lines according to an embodiment of the present invention.
FIG. 2 is a graph showing the results of RT-PCR for the amount of HOXB5 expression in 5 non-cancerous and 5 cancerous breast tissues according to an embodiment of the present invention.
FIG. 3 is a diagram showing the classification of HOXB5 immunohistochemical staining results in breast tissue according to an embodiment of the present invention.
4 is a graph showing the results of analysis of HOXB5 expression and breast cancer prognosis according to an embodiment of the present invention.
FIG. 5 is a graph showing the results of analysis of cancer proliferative activity in a breast cancer cell line inhibited by HOXB5 expression according to an embodiment of the present invention.
FIG. 6 is a graph showing the results of analysis of cancer proliferation in HOXB5 overexpressed breast cancer cell lines according to an embodiment of the present invention. FIG.
FIG. 7 is a graph showing morphological changes in HOXB5 overexpressed breast cancer cells according to an embodiment of the present invention. FIG.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example  1: in breast cancer cell lines and tissues HOXB5  Investigation of the expression level

Example  1.1. In breast cancer cell lines and tissues HOXB5 of mRNA  Expression level analysis

To demonstrate the association of breast cancer with HOXB5, five of each of the nine breast-derived cell lines and breast cancer tissue / non-cancerous breast tissue were isolated, and total RNA was isolated using Trizol. RT-PCR was performed to express HOXB5 The amount was confirmed.

The expression of HOXB5 was examined in nine types of breast cancer cell lines (FIG. 1) and five non-cancerous and cancerous breast tissues (FIG. 2), respectively. As a result, HOXB5 expression was found to be high in estrogen receptor positive breast cancer cell lines and tissues Respectively. The characteristics related to the estrogen receptor, progesterone receptor and human epidermal growth factor (HER2) of breast cancer cell lines and non-cancerous and cancerous breast tissues used in the Examples are shown in Table 1 and Table 2, respectively .

Cell line Estrogen receptor Progesterone receptor Human epidermal growth factor 2 MCF10A voice voice voice BT474 positivity positivity positivity MCF7 positivity positivity voice T47D positivity positivity voice HCC1428 positivity positivity voice MDA-MB-453 voice voice voice SKBR3 voice voice positivity MDA-MB-231 voice voice voice HCC70 voice voice voice

group focus Estrogen receptor Progesterone receptor Human epidermal growth factor 2 N1 Adenosine
(Adenosis tumor) N2 Fibroadenoma
(Fibroadenoma) N3 normal
(Normal) N4 Papilloma
(Papillomatosis) N5 Papilloma
(Papilloma) T1 Invasive duct carcinoma
(IDC) > 50% > 50% voice T2 Invasive duct carcinoma
(IDC) > 50% > 50% Weak positive T3 Invasive duct carcinoma
(IDC) > 50% 10-50% voice T4 Invasive duct carcinoma
(IDC) <10% <10% Weak positive T5 Invasive duct carcinoma
(IDC) voice voice Strong positive N: non-invasive human breast tissue (non-malignant human breast tissue)
T: human breast cancer tissue
IDC: invasive ductal carcinoma

Example  1.2. In breast cancer cell lines and tissues HOXB5 Of protein expression

Based on the results of Example 1.1, a tissue microarray (TMA) containing 63 breast tissues and 34 normal breast tissues was prepared for the analysis of a larger number of tissue samples, and immunohistochemical staining we investigated the expression of HOXB5 protein in the tissues by immunohistochemistry and analyzed the breast cancer types and clinicopathological factors of the tissues. According to the expression level of HOXB5, it was classified as negative, weak positive, intermediate positive and strong positive, and representative coloring degree of each was shown in [FIG. 3]. The total number of tissues was classified according to two criteria, and the results were shown in Table 3. Criterion 1 was classified as negative and positive (including mildly positive, intermediate positive and strong positive) according to HOXB5 expression, and criterion 2 was classified as negative and positive And a strong positive group. As a result, HOXB5 expression was significantly higher in breast cancer tissues than in normal breast tissues in both the criteria 1 and the criteria 2.

standard HOXB5  Expression normal Breast cancer Standard Deviation One

voice
positivity
(Weak, medium, strong) 27 (73.5%)
9 (26.5%)
28 (45.2%)
34 (54.8%)

0.008
2
Negative and weak positive
Intermediate positive and strong positive 34 (100%)
0 (0%) 54 (87.1%)
8 (12.9%)
0.047

Example  1.3. HOXB5  Analysis of relationship between expression and clinicopathologic factors

Using the online database (www.kmplot.com), we analyzed the degree of HOXB5 expression and the prognosis of breast cancer patients. Survival analysis was performed using an online database Kaplan Meier Plotter because the number of samples included in the tissue microarray was not sufficient to analyze the correlation between the type of cancer and the expression of HOXB5 (Www.kmplot.com).

The expression of HOXB5 was correlated with the estrogen receptor status (ER status), and the expression of HOXB5 was higher in estrogen receptor - positive breast cancer tissues than in estrogen receptor - negative ones (Table 4). There was no correlation with other clinical factors.

variable HOXB5  voice
(n = 28) HOXB5  positivity
(n = 33) Standard Deviation age
(Age) 48.4 ± 12.5 50.4 ± 12.9 0.542 Tumor size
(Tumor size) ≤ 2 cm 3 (10.7%) 3 (9.1%) 1,000 > 2 cm 25 (89.3%) 30 (90.9%) Nodular state
(Node status) Vision 13 (46.4%) 15 (45.5%) 0.939 transition 15 (53.6%) 18 (54.5%) step
(Stage) Ⅰ and Ⅱ 19 (67.9%) 26 (78.8%) 0.333 Ⅲ and Ⅳ 9 (32.1%) 7 (21.2%) Histological stage
(Histologic grade) Ⅰ 6 (24.0%) 8 (24.2%) 0.983 Ⅱ and Ⅲ 19 (76.0%) 25 (75.8%) Estrogen receptor status
(ER status) voice 16 (57.1%) 10 (30.3%) 0.035 positivity 12 (42.9%) 23 (69.7%) Progesterone receptor status
(PR status) voice 7 (25.0%) 8 (24.2%) 0.945 positivity 21 (75.0%) 25 (75.8%) Human epidermal growth factor 2
(HER2 status) voice 19 (67.9%) 26 (78.8%) 0.333 positivity 9 (32.1%) 7 (21.2%)

In addition, the relationship between HOXB5 expression and breast cancer prognosis was analyzed and shown in Fig. HOXB5 was not associated with distant metastasis free survival (DMFS) in all patients (Fig. 4- (a)), but survival without distant metastasis was seen only in patients with estrogen receptor And the prognosis is poor when HOXB5 expression is high (Fig. 4- B). In patients with histologic grade (HG) I who were positive for estrogen receptors and lymph node metastasis, the higher the expression of HOXB5, the worse the prognosis (Fig. 4-) High expression of HOXB5 was associated with poor endocrine responsiveness when viewed from the overall survival (OS) (Figure 4-4).

Example  2. Breast cancer cell line HOXB5 Functional study of

We investigated the role of HOXB5 in the development and progression of breast cancer by investigating the proliferation, apoptosis, invasiveness and colony forming ability of breast cancer cells after inducing HOXB5 overexpression or expression inhibition in breast cancer cell lines.

Example  2.1. HOXB5  Inhibition of expression and induction of overexpression cell line

Among the estrogen receptor-positive breast cancer cell lines, BT474, MCF-7, T47D and SKBR3 and MDA MB 231 were used among estrogen receptor-negative cell lines. MCF10A was used as the normal breast cell line. MCF-7, BT474, and MDA MB 231 were cultured in DMEM medium supplemented with 10% bovine serum (FBS) and T47D medium supplemented with RPMI 1640 medium. SKBR3 was used in McCoy's 5a medium and MCF10A was supplemented with 5% horse serum, 20 ng / ml endothelial growth factor (EGF), and 0.5 μg / ml hydrocortisone in DMEM / F12 medium , Cholera toxin (100 ng / ml) and insulin (10 μg / ml). The siRNA for the inhibition of expression was commissioned by GENORUS and shRNA was purchased from Thermo. After transient effect by siRNA or introduction of shRNA, antibiotics selection process was performed to establish a cell line in which a specific gene was knocked down. The expression of the gene was inhibited by RT-PCR and Western blotting. Conversely, for overexpression, cloning is performed in an expression vector of pCDNA3-HA to include the entire coding sequence of the gene of interest, followed by transfection to confirm overexpression, To obtain a cell line stably expressing the gene.

Example  2.2. Breast cancer and Non-cancerous  Tissue Microarrays in Breast Tissue TMA Production

Human breast tissue was obtained from Ilsan Hospital after signing a consent form for clinical study from patients who underwent breast cancer surgery between June and July 2012. Immediately after surgery, it was placed in RNA storage solution and stored in a -80 ℃ freezer until use . For tissue embedded in paraffin fixed to formalin, tissue microarray (TMA) was prepared using tissue-arraying instrument (AccuMax Array; Petagen Inc., Seoul, Korea).

Example  2.3. In breast cancer cell lines and breast cancer tissues RNA  Separation and Real - time PCR

RNA was extracted from breast cancer cell lines using Trizol-Reagent (Invitrogen) and cDNA was synthesized using Impron-II RT reverse transcriptase reagent. The PCR was carried out basically at 95 ° C for 2 minutes, followed by 30 cycles of 95 ° C for 30 seconds, 55-58 ° C for 20 seconds, and 72 ° C for 30 seconds, and for each gene, And the number of cycles. Beta-actin or GAPDH was used as a control. Human breast tissue was obtained by pulverizing tissue using liquid nitrogen in a mortar and then extracting total RNA using the same method as described above and synthesizing cDNA and using it for real-time PCR. Real-time PCR was performed on an Applied Biosystems 7300 Real-Time PCR System instrument using a Master Mix (Power SYBR Green PCR Master Mix, Applied Biosystems) reagent. Data were analyzed using software from Applied Biosystems. Fluorescent output was converted to the potency (Ct) and analyzed using the sample ratio to normal (2-ΔΔCt).

Example  2.4. Western Western blotting and  Immunohistochemical staining ( Immunohistochemistry )

Cells were resuspended in RIPA buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% DSD, 0.5% sodium deoxycholate, 1% Triton X-100 and proteinase inhibitor cocktail ) To make a protein lysate. Protein was quantified using a BCA (bicinchoninic acid) protein assay kit (Thermo, # 23227). Proteins were separated by gel electrophoresis and then transferred to PVDF membrane. After blocking with 5% skim milk, the first antibody capable of detecting each protein was added and incubated overnight at 4 ° C. After the 2nd antibody incubation, the resultant was subjected to a color reaction using a reagent of SuperSignal West Pico Chemiluminescent Substrate (Pierce Cat # 34080) manufactured by Pierce, and then sensitized. Immunohistochemistry In the experiment, cells were blocked with normal goat serum (3% in PBST, 0.05% Tween 20) after treatment with Vector untransfected antigen unmasking solution. Tissue sections were incubated with the antibody and detected with a secondary antibody conjugated with Alexa Fluor. Fluorescence images were observed with an Olympus fluorescence microscope (Olympus IX70).

Example  2.5. Cell proliferation Cell proliferation  )and Cell death ( Apoptosis ) analysis

The survival rate of the cells was measured using a reagent such as 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) reagent. 20 μl of MTT solution in PBS (5 mg / ml) was added to the cells and incubated at 37 ° C for 3.5 hours. The reaction was stopped with a stop solution (4 mM HCl, 0.1% Nondet P-40 (NP40) in isopropanol) and the absorbance at 560 nm was measured. Apoptosis was performed by staining adenosine V-FITC and propidium iodide using flow cytometry using an Annexin V-FITC kit (Kombiotech) Was performed.

Example  2.6. anchorage Independent Colony  formation( anchorage - independent colony  formation Light cloth for  analysis( Soft agar assay )

Existing agar medium was used as agarose final concentration 0.5%, and upper agar medium was used as final concentration 0.35%. The cells to be analyzed were prepared so that 5 x 10 ³ cells were present in 100 μl of the culture medium and transplanted onto the base agar medium. After incubation in a 37 ° C incubator for 15-21 days, colonies were observed by staining with 0.5 mg / ml of Nitroblue Tetrazolium (NBT).

Example  2.7. Matrigel  Invasive Analysis Matrigel invasion assay )

Matrigel ™ (BD) was mixed in a coating buffer (0.01 M Tris, pH 8.0 and 0.7% NaCl) to a final concentration of 300 μg / ml, and then placed in a 24-well plate 100 μl was added to the individual insert. The cells were transplanted into an insert plate prepared so that 5 x 10 ⁴ cells were contained in 500 μl serum-free medium. 750 μl of a culture medium containing 10% bovine serum (FBS) was added to the lower chamber outside the insertion plate. After incubation at 37 ° C for an appropriate period of time (1 to 3 DAY), the number of cells infiltrating through the matrigel (cells attached to the bottom surface of the insert) was determined by DAPI staining. The obtained images were analyzed using image analysis software.

Example  2.8. HOXB5  Effect by inhibition of expression

A lentiviral vector expressing shRNA against HOXB5 was infected with estrogen receptor-positive breast cancer cell line T47D by the methods of Examples 2.1 to 2.7 to inhibit HOXB5 expression in breast cancer cell lines (# 1, # 2, # 3) (FIG. 5-a), protein expression level (FIG. 5-b), cell proliferation effect (FIG. 5-c) and colony forming ability . These cell lines showed decreased cell proliferation and decreased colony forming ability.

Example  2.9. HOXB5  Effect of Overexpression

HOXB5-overexpressed breast cancer cell lines (# 1, # 2, # 3) overexpressing HOXB5 in the estrogen receptor-positive breast cancer cell line MCF7 were prepared by the methods of Examples 2.1 to 2.7 and their mRNA expression levels (Fig. 6), the cell proliferation effect (Fig. 6), the colony forming ability (Fig. 6) and the cancer cell infiltration ability (Fig. In this case, contrary to the inhibition of HOXB5 expression in Example 2.8, it showed an increase in cell proliferation, an increase in colony forming ability, and an increase in invasiveness of cancer cells.

In particular, the MCF7 cell line overexpressing HOXB5 induces a morphological change in cell morphology as compared to a cell line having low expression level, as shown in Fig. The MCF7 cell line overexpressing HOXB5 plays an important role in the metastasis of cancer cells, such as a decrease in the epithelial marker E-cadherin, an increase in the mesenchymal marker Vimentin, and a change in the expression position of β-catenin Epithelilal-mesenchymal transition (EMT) occurs. These results show that HOXB5 plays an important role in the development and progression of breast cancer.

<110> Yonsei Industry Academic Cooperation Foundation <120> Use of HOXB5 for diagnosis and treatment of breast cancer <130> DPB140015 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 1830 <212> DNA <213> human <400> 1 gtgaagcaca gggttataac gaccacgatc cacaaatcaa gccctccaaa atcacccaaa 60 tgagctcgta ctttgtaaac tccttctcgg ggcgttatcc aaatggcccg gactatcagt 120 tgctaaatta tggcagtggc agctctctga gcggctctta cagggatccc gctgccatgc 180 acaccggctc ttacggctac aattacaatg ggatggacct cagcgtcaac cgctcctcgg 240 cctcctccag ccactttggg gcggtgggcg agagctcgcg cgccttcccc gcgcccgccc 300 aggagccccg cttcaggcaa gcggcttcga gctgctccct gtcctcgccc gagtccctgc 360 cctgcaccaa cggcgacagc cacggcgcca agccctctgc ttcgtccccc tccgaccagg 420 cgacctcagc cagctccagc gccaatttca ccgaaataga cgaggccagc gcgtcctcgg 480 agcctgagga agcggcaagc cagctaagca gccccagcct agctcgggcg cagccagagc 540 ccatggccac ctccacagcc gcgcccgagg ggcagactcc gcaaatattc ccctggatga 600 ggaagcttca catcagccat gatatgaccg ggccggacgg gaaaagggcc cggaccgcgt 660 atacccgcta ccagaccctg gagctggaaa aggagttcca cttcaaccgc tacctgaccc 720 ggcgacggcg catcgagatc gcccacgcac tctgcctgtc cgagcgccag atcaagatct 780 ggttccagaa ccggcgcatg aagtggaaga aggacaacaa attgaaaagt atgagcctgg 840 ctacagctgg cagcgccttc cagccctgag cccgcccaga ggagcccagc ggcccaagag 900 cccgtgccac ccccagccct ggcccctcca atcctccccg ctctgccgcc gcccgctggg 960 gaccggttcc cacaagcctg cctcgccttg tgttacgata tttcgtttgg tcttaggtct 1020 tcctgtggct ccctctctcc tggactggtt atcttgttat tattgttaat aataattatt 1080 attattattt tccttccatg ctcccaactc ccttctgctt gtcccaaatc cgccagtgtt 1140 tctgaatgtt tgtgtctgtg gttgcagtct ttcccccagg aaaaaaaaaa aaagaaattc 1200 gcatgtttaa tgtgaactct cccctcccca tctgtgttct aacttattta taaaaagatg 1260 atcgctgtat tttgagtttc agctggaaac ttctgtaagg ggcagcagtt gaggtggggt 1320 agtgccgcag tggggtcaag ctgagctggc ttcggagatg gagtcccttt tcattctcct 1380 cctcctccct cctcactccc taggcccaag tctcctaggg gcttggtcct agggtgggaa 1440 ggggctaggg aggaccaaag ggatggtatt gagaagagag aaagaagta gtgagattta 1500 agttcctgct gcctgggtag gccccacaag gcctggtctg ggagtatacg gaaacaaaaa 1560 tgatcctcag tgcaaaatgt cttgtgtatt tctctgtgaa tccatgggtc tggctagagg 1620 gcccaaagct tgtaaatatg gggatagtct gggtcagacc catctctccc ttacccatct 1680 tgcttccaag accatttgta gtgagcgagt ggatgctgtg ctacgtgtga aatctgtctt 1740 tgcggggcct gtctcagtga ttcgcttttg gtatttgttt gtagctttcc tggaagtcaa 1800 ataaatgttt cccccactcc aaaaaaaaaa 1830 <210> 2 <211> 269 <212> PRT <213> human <400> 2 Met Ser Ser Tyr Phe Val Asn Ser Phe Ser Gly Arg Tyr Pro Asn Gly   1 5 10 15 Pro Asp Tyr Gln Leu Leu Asn Tyr Gly Ser Gly Ser Ser Leu Ser Gly              20 25 30 Ser Tyr Arg Asp Pro Ala Ala Met His Thr Gly Ser Tyr Gly Tyr Asn          35 40 45 Tyr Asn Gly Met Asp Leu Ser Val Asn Arg Ser Ser Ala Ser Ser Ser      50 55 60 His Phe Gly Ala Val Gly Glu Ser Ser Arg Ala Phe Pro Ala Pro Ala  65 70 75 80 Gln Glu Pro Arg Phe Arg Gln Ala Ala Ser Ser Cys Ser Leu Ser Ser                  85 90 95 Pro Glu Ser Leu Pro Cys Thr Asn Gly Asp Ser His Gly Ala Lys Pro             100 105 110 Ser Ala Ser Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala         115 120 125 Asn Phe Thr Glu Ile Asp Glu Ala Ser Ala Ser Ser Glu Pro Glu Glu     130 135 140 Ala Ala Ser Gln Leu Ser Ser Pro Ser Leu Ala Arg Ala Gln Pro Glu 145 150 155 160 Pro Met Ala Thr Ser Thr Ala Ala Pro Glu Gly Gln Thr Pro Gln Ile                 165 170 175 Phe Pro Trp Met Arg Lys Leu His Ile Ser His Asp Met Thr Gly Pro             180 185 190 Asp Gly Lys Arg Ala Arg Thr Ala Tyr Thr Arg Tyr Gln Thr Leu Glu         195 200 205 Leu Glu Lys Glu Phe His Phe Asn Arg Tyr Leu Thr Arg Arg Arg Arg     210 215 220 Ile Glu Ile Ala His Ala Leu Cys Leu Ser Glu Arg Gln Ile Lys Ile 225 230 235 240 Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys Asp Asn Lys Leu Lys                 245 250 255 Ser Met Ser Leu Ala Thr Ala Gly Ser Ala Phe Gln Pro             260 265

Claims (23)

A pharmaceutical composition for inhibiting angiogenesis, comprising at least one selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific for the HOXB5 gene as an effective ingredient for inhibiting the expression of HOXB5 (Homeobox B5) gene. The method according to claim 1,
Wherein the inhibition of angiogenesis is inhibition of breast cancer growth. 3. The method of claim 2,
Breast cancer is a positive estrogen receptor. The method according to claim 1,
The HOXB5 (Homeobox B5) gene is represented by SEQ ID NO: 1. A pharmaceutical composition for inhibiting angiogenesis, comprising an antibody specific for HOXB5 as an active ingredient for HOXB5 (Homeobox B5) protein inhibition. 6. The method of claim 5,
Wherein the inhibition of angiogenesis is inhibition of breast cancer growth. The method according to claim 6,
Breast cancer is a positive estrogen receptor. 6. The method of claim 5,
Wherein the HOXB5 protein is SEQ ID NO: 2. A pharmaceutical composition for inhibiting cancer metastasis comprising at least one selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific for the HOXB5 gene as an effective ingredient for inhibiting the expression of HOXB5 (Homeobox B5) gene. 10. The method of claim 9,
Wherein the inhibition of cancer metastasis is inhibition of breast cancer growth. 11. The method of claim 10,
Breast cancer is a positive estrogen receptor. HOXB5 (Homeobox B5) A pharmaceutical composition for inhibiting cancer metastasis comprising HOXB5 specific antibody as an active ingredient for protein inhibition. 13. The method of claim 12,
Wherein the inhibition of cancer metastasis is inhibition of breast cancer growth. 14. The method of claim 13,
Breast cancer is a positive estrogen receptor. A composition for diagnosing estrogen receptor positive (ER +) type breast cancer comprising a primer or a probe that specifically binds to mRNA of HOXB5 (Homeobox B5) gene. 16. The method of claim 15,
The HOXB5 (Homeobox B5) gene is the one represented by SEQ ID NO: 1. HOXB5 (Homeobox B5) An estrogen receptor positive (ER +) type breast cancer diagnostic composition comprising a HOXB5 specific antibody as an active ingredient for protein inhibition. 18. The method of claim 17,
Wherein the HOXB5 protein is SEQ ID NO: 2. The level of mRNA of the HOXB5 (Homeobox B5) gene of the subject was measured with a primer or a probe that specifically binds to the mRNA of HOXB5 (Homeobox B5) gene to determine the estrogen receptor positive (ER +) type / RTI &gt; 20. The method of claim 19,
Wherein the HOXB5 (Homeobox B5) gene provides the information represented by SEQ ID NO: 1. HOXB5 (Homeobox B5) A method for providing information on estrogen receptor positive (ER +) type breast cancer diagnoses, including HOXB5 specific antibodies as active ingredients for protein inhibition. 22. The method of claim 21,
Wherein the HOXB5 protein provides the information represented by SEQ ID NO: 2. When the expression level of the HOXB5 (Homeobox B5) gene or the expression level of the protein encoded by the gene is suppressed after the administration of a candidate substance for estrogen receptor positive (ER +) type breast cancer treatment, the candidate substance is subjected to estrogen receptor- A method for screening estrogen receptor positive (ER +) type breast cancer therapeutic agent, which is determined to be an estrogen receptor positive (ER +) type breast cancer therapeutic agent.

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