KR20140079255A - A composition and kit for diagnosing a breast cancer including a polynucleotide within a vesicle, and a method for diagnosing a breast cancer using the same - Google Patents

Abstract

A breast cancer can be diagnosed early with a minimum invasive method and high convenience of a patient using a composition and a kit for diagnosing the breast cancer including: one or more micro RNAs which are selected from a group comprising hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR-103, hsa-miR-142-3p, hsa-miR-144, hsa-miR-15a, hsa-miR-185, hsa-miR-93, and hsa-miR-30c contained in a vesicle; or polynucleotide which is in the same or in complementary relations with a fragment of the micro RNA. Also, the breast cancer can be diagnosed using a method for diagnosing the breast cancer and a method for obtaining information required for the diagnosis of the breast cancer using the composition and the kit.

Description

Technical Field [0001] The present invention relates to a composition and a kit for diagnosing breast cancer comprising polynucleotides in vesicles, and a method for diagnosing breast cancer using the same, and a kit for diagnosing breast cancer using the polynucleotide,

A composition and a kit for diagnosing breast cancer comprising polynucleotides in vesicles, and a method for diagnosing breast cancer using the same.

Micro vesicles in vivo are small vesicles of membrane structure that are present in various kinds of cells or secreted from cells. The microvesicles secreted out of the cell are (i) exosomes: membranous vesicles of 30-100 nm in diameter originated from the origin of the bacteria, (ii) shedding microvesicles (SMVs): flowing directly from the plasma membrane (Iii) Apoptotic blebs: vesicles of 50-5000 nm in diameter, which are released by dying cells.

Among them, exosome is a small vesicle of membrane structure secreted from various kinds of cells. The diameter of the exosome may be approximately 30 to 100 nm. Exosomes have been observed to be released and secreted out of the cell, originating from specific compartments within the cell called multivesicular bodies (MVBs), rather than being directly separated from the plasma membrane in electron microscopic studies. That is, when fusion of the polycation and the plasma membrane occurs, such vesicles are released into the extracellular environment, which is called exosomes. It is unclear how these exosomes are produced by molecular mechanisms, but it is possible that not only red blood cells but also various types of immune cells and tumor cells, including B-lymphocytes, T-lymphocytes, dendritic cells, platelets and macrophages, It is known to produce and secrete exosome in the state of being. Exosomes are known to be released from many different cell types under normal, pathological, or both conditions.

In addition, exosomes may comprise microRNAs (miRNAs). The miRNA can be used to confirm the status of the cell or organ containing it. The condition may be a disease, for example a neurological disease such as cancer, hereditary disease, heart disease, or schizophrenia.

Diagnosis of breast cancer through conventional biopsy is an invasive method, which is accompanied by patient's suffering, difficulty in frequent screening, and high cost. In addition, there is no accurate blood protein marker for the diagnosis of breast cancer through conventional blood tests, and circulating tumor cells (CTC) can be applied only to metastatic breast cancer, which is not applicable to early cancer diagnosis .

Therefore, it is necessary to screen for markers in breast cancer-specific blood for early diagnosis of breast cancer in a minimally invasive manner.

There is provided a composition for diagnosing breast cancer.

A kit for diagnosing breast cancer is provided.

Provide a method for diagnosing breast cancer.

Provides a way to get the information needed to diagnose breast cancer.

HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- one or more microRNAs (miRNA) selected from the group consisting of miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary to the fragments thereof A composition for diagnosing breast cancer is provided.

The vesicle or vesicle refers to a membrane structure surrounded by a lipid bilayer. For example, the vesicle may be microbejac or exosomes. Microvesicles are small vesicles of membrane structure derived from cells. Microbeads are used interchangeably with circulating microvesicles or microparticles. Microbeads can be present in cells or secreted from cells. Microvessels secreted out of the cell can be exosomes, also known as shedding microvesicles (SMVs), or apoptotic blebs. The exosomes may be membranous beads of 30 to 100 nm in diameter originating from a plant source. The etomome can be drained directly from the plasma membrane and be a large filmy bezuge of 50-1,000 nm diameter. The cell suicide blister may be a bezacl with a diameter of 50 to 5,000 nm, which is discharged by dying cells. In vivo microvesicles may include microRNA (miRNA) or mRNA (messenger RNA).

The microRNA (miRNA) refers to short ribonucleic acid found in eukaryotic cells. MicroRNAsms are responsible for regulating gene expression in vivo and are about 17 to about 25 nucleotides in length (hereinafter referred to as "nt "), which are encoded by DNA. They are transcribed from the genome and then cut into fragments to increase or decrease the expression of specific proteins. To date, known functions of microRNAs in mammals include insulin secretion regulation, lymphocyte differentiation regulation, cell division regulation, cell death control, and viral replication regulation. MiRNAs such as hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- , hsa-miR-15a, hsa-miR-185, hsa-miR-93, hsa-miR-30c or a combination thereof. HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR-103, hsa- hsa-miR-185, hsa-miR-93, and hsa-miR-30c correspond to human sapiens miR-126, human miR-23a, human miR-24, human miR- -142-3p, human miR-144, human miR-15a, human miR-185, human miR-93 and human miR-30c. hsa-miR-126 has, for example, the nucleotide sequence of SEQ ID NO: 1. hsa-miR-23a has, for example, the nucleotide sequence of SEQ ID NO: 2. hsa-miR-24 has, for example, the nucleotide sequence of SEQ ID NO: 3. hsa-miR-19b has, for example, the nucleotide sequence of SEQ ID NO: 4. hsa-miR-103 has, for example, the nucleotide sequence of SEQ ID NO: 5. hsa-miR-142-3p has, for example, the nucleotide sequence of SEQ ID NO: 6. hsa-miR-144 has, for example, the nucleotide sequence of SEQ ID NO: 7. hsa-miR-15a has, for example, the nucleotide sequence of SEQ ID NO: 8. hsa-miR-185 has, for example, the nucleotide sequence of SEQ ID NO: 9. hsa-miR-93 has, for example, the nucleotide sequence of SEQ ID NO: 10. hsa-miR-30c has, for example, the nucleotide sequence of SEQ ID NO: 11.

The fragment refers to a polynucleotide having a contiguous base sequence of microRNA. The length of the fragment may be, for example, from 2 nt to 25 nt, from 3 nt to 24 nt, from 4 nt to 23 nt, from 5 nt to 22 nt, from 6 nt to 21 nt, from 7 nt to 20 nt, 9 nt to 18 nt, 10 nt to 17 nt, 11 nt to 16 nt, 12 nt to 15 nt, or 13 nt to 14 nt.

The polynucleotide may be, for example, one having the same base sequence as the microRNA or a fragment thereof. The polynucleotide may be, for example, one having a base sequence complementary to the microRNA or a fragment thereof. The polynucleotide may be a primer or a probe. The length of the polynucleotide may be, for example, from 2 nt to 25 nt, from 3 nt to 24 nt, from 4 nt to 23 nt, from 5 nt to 22 nt, from 6 nt to 21 nt, from 7 nt to 20 nt, , 9 nt to 18 nt, 10 nt to 17 nt, 11 nt to 16 nt, 12 nt to 15 nt, or 13 nt to 14 nt.

The composition may further comprise a substance capable of binding specifically to the vesicle or interfering with the lipid bilayer of vesicles. A substance that can specifically bind to the vesicle may be, for example, a substance capable of binding to the surface protein, lipid, or sugar of the vesicle. The surface proteins may be, for example, CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2 or a combination thereof. Substances capable of specifically binding to the vesicles include, for example, substances having a binding affinity to a protein, a substrate of an enzyme, a coenzyme, a regulator, a substance that specifically binds to a receptor, a lectin, a sugar, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, in addition to a protein, an antigen, an antibody or an antigen binding fragment thereof, a hormone, The antigen-binding fragment comprises an antigen binding site, for example a single-domain antibody, Fab, Fab 'or scFv. The material that can interfere with the lipid bilayer of the vesicle may be, for example, a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. The lipophilic moiety may be, for example, a fatty acid, a sterol, or a glyceride. The amphiphilic moiety may be, for example, a phospholipid or a sphingolipid. The amphoteric ion moiety may be, for example, sulfobetaine, carboxybetaine, or phosphorylcholine. Substances capable of specifically binding to the vesicle or interfering with the lipid bilayer of vesicles may be associated with a solid support. The solid support may be, for example, a polystyrene plate, a polystyrene bead, or a magnetic bead.

HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- one or more microRNAs (miRNA) selected from the group consisting of miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary to the fragments thereof A kit for diagnosing breast cancer is provided.

The vesicles, microRNAs, fragments, and polynucleotides are as described above.

The kit may further comprise a material capable of binding specifically to the vesicle or interfering with the lipid bilayer of vesicles. A substance that can specifically bind to the vesicle may be, for example, a substance capable of binding to the surface protein, lipid, or sugar of the vesicle. The surface proteins may be, for example, CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2 or a combination thereof. Substances capable of specifically binding to the vesicles include, for example, substances having a binding affinity to a protein, a substrate of an enzyme, a coenzyme, a regulator, a substance that specifically binds to a receptor, a lectin, a sugar, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, in addition to a protein, an antigen, an antibody or an antigen binding fragment thereof, a hormone, The antigen-binding fragment comprises an antigen binding site, for example a single-domain antibody, Fab, Fab 'or scFv. The material that can interfere with the lipid bilayer of the vesicle may be, for example, a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. The lipophilic moiety may be, for example, a fatty acid, a sterol, or a glyceride. The amphiphilic moiety may be, for example, a phospholipid or a sphingolipid. The amphoteric ion moiety may be, for example, sulfobetaine, carboxybetaine, or phosphorylcholine. Substances capable of specifically binding to the vesicle or interfering with the lipid bilayer of vesicles may be associated with a solid support. The solid support may be, for example, a polystyrene plate, a polystyrene bead, or a magnetic bead.

Separating the vesicles from the biological sample separated from the subject;

Wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa miR-103, hsa-miR-142, hsa-miR-103, hsa-miR-142 and hsa-miR- Or more microRNA;

Comparing the amount of microRNAs measured with the amount of microRNAs measured from the vesicles isolated from the control sample; And

Determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample.

The method comprises separating the vesicles from the biological sample separated from the subject.

An individual may be, for example, a mammal, including a human.

The biological sample may be, for example, urine, mucus, saliva, tears, blood, plasma, serum, sputum, spinal fluid, pleural effusion, aspiration nipple, lymphatic fluid, airway fluid, intestinal fluid, urinary fluid, milk, , Intracapsular fluid, ascites fluid, cystic tumor fluid, positive fluid, or a combination thereof.

The vesicles are as described above.

The step of separating the vesicles from the biological sample may be carried out by, for example, separation using a solid support or centrifugal force, density gradient method, ultracentrifugation, filtration, dialysis, immunoaffinity chromatography using antibodies, Can be performed in one way. The step of separating the vesicles from the biological sample can comprise, for example, incubating the vesicles with a substance capable of binding to the lipid bilayer of a substance or vesicle capable of specifically binding to the vesicle. The incubation can be performed in vitro. The step of separating the vesicles from the biological sample may comprise, for example, washing. A substance that can specifically bind to the vesicle may be, for example, a substance capable of binding to the surface protein, lipid, or sugar of the vesicle. The surface proteins may be, for example, CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2 or a combination thereof. Substances capable of specifically binding to the vesicles include, for example, substances having a binding affinity to a protein, a substrate of an enzyme, a coenzyme, a regulator, a substance that specifically binds to a receptor, a lectin, a sugar, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, in addition to a protein, an antigen, an antibody or an antigen binding fragment thereof, a hormone, The antigen-binding fragment comprises an antigen binding site, for example a single-domain antibody, Fab, Fab 'or scFv. The material that can interfere with the lipid bilayer of the vesicle may be, for example, a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. The lipophilic moiety may be, for example, a fatty acid, a sterol, or a glyceride. The amphiphilic moiety may be, for example, a phospholipid or a sphingolipid. The amphoteric ion moiety may be, for example, sulfobetaine, carboxybetaine, or phosphorylcholine. Substances capable of specifically binding to the vesicle or interfering with the lipid bilayer of vesicles may be associated with a solid support. The solid support may be, for example, a polystyrene plate, a polystyrene bead, or a magnetic bead.

The method comprises measuring the amount of microRNA (miRNA) contained in the separated vesicles, wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR- Hsa-miR-93, and hsa-miR-30c, hsa-miR-103, hsa-miR-142-3p, hsa-miR-144, hsa-miR-15a, hsa-miR- Wherein the microRNA is one or more microRNAs.

The microRNAs are as described above.

The amount of the microRNA is preferably selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- a microRNA (miRNA) selected from the group consisting of -mR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary thereto It can be measured using primer or probe. The length of the polynucleotide may be, for example, from 2 nt to 25 nt, from 3 nt to 24 nt, from 4 nt to 23 nt, from 5 nt to 22 nt, from 6 nt to 21 nt, from 7 nt to 20 nt, , 9 nt to 18 nt, 10 nt to 17 nt, 11 nt to 16 nt, 12 nt to 15 nt, or 13 nt to 14 nt. The amount of the microRNA can be measured by, for example, quantitative reverse transcription-polymerase chain reaction (qRT-PCR).

The method includes comparing the amount of microRNAs measured to the amount of microRNAs measured from the vesicles isolated from the control sample.

The control sample may be a sample derived from a normal or benign tumor patient.

The method includes determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample.

The method may further comprise dissolving the vesicles separated from the biological sample. The dissolution of the vesicles may be carried out, for example, in the presence of a solvent comprising a chaotropic salt, an organic solvent or a surfactant. The dissolution of the vesicles can be carried out, for example, by heating, stirring, rotating, vortexing, or a combination thereof.

Separating the vesicles from the biological sample separated from the subject;

Wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa miR-103, hsa-miR-142, hsa-miR-103, hsa-miR-142 and hsa-miR- Or more microRNA;

Comparing the amount of microRNAs measured with the amount of microRNAs measured from the vesicles isolated from the control sample; And

Determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample.

The method comprises separating the vesicles from the biological sample separated from the subject.

The subject, biological sample, vesicle and separation are as described above.

The method comprises measuring the amount of microRNA (miRNA) contained in the separated vesicles, wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR- Hsa-miR-93, and hsa-miR-30c, hsa-miR-103, hsa-miR-142-3p, hsa-miR-144, hsa-miR-15a, hsa-miR- Wherein the microRNA is one or more microRNAs.

The amounts of the microRNA and the microRNA are as described above.

The method includes comparing the amount of microRNAs measured to the amount of microRNAs measured from the vesicles isolated from the control sample.

The sample and the control sample are as described above.

The method includes determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample.

The method may further comprise dissolving the vesicles separated from the biological sample. The dissolution of the vesicles is as described above.

HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- a composition for breast cancer diagnosis comprising at least one microRNA selected from the group consisting of miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary thereto, By using the kit, the method of diagnosing breast cancer and the method of acquiring the information necessary for diagnosis of breast cancer, it is possible to diagnose the breast cancer at a minimally invasive manner and improve the convenience of the patient.

Figure 1 shows the amount of microbicycles in benign tumor patients and breast cancer patients when the microbicycles were isolated from the CD83 protein.
Figures 2a and 2b show differences in the amount of microRNAs in benign tumor patients and breast cancer patients.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1. Breast Cancer Related Microbesic  Surface protein Marker  Selection

Blood was collected from 20 benign tumor carriers (benign tumor patients), not breast cancer, and 22 patients with luminal type B breast cancer (breast cancer patients), and plasma was obtained from the blood. The obtained plasma was centrifuged at 3,000 g for 5 minutes at 4 DEG C to remove plasma lipids and cell debris.

An anti-CD83 antibody (BD Pharmingen) was added to the plasma and immunoprecipitated to isolate the microvesicles having the CD83 surface protein in plasma. To determine the amount of CD83-expressing microvesicles isolated in benign tumor bearing and breast cancer patients, the amount of CD9 protein, a microvessel marker, was confirmed by Western blotting. Western blotting was performed using an anti-CD9 antibody (Novus Biologicals).

The obtained microbicycles were subjected to western blotting to select CD83, ERBB2 and FASL proteins having a significant difference in positive tumor patients and breast cancer patients.

Microvessels were obtained with anti-CD83 antibody (BD Pharmingen), anti-ERBB2 antibody (R & D Systems) and anti-FASL (BD Pharmingen) antibody and the amount of microbicide obtained was confirmed by Western blotting. Western blotting used an anti-CD9 antibody (Novus Biologicals).

The difference in the amount of protein was determined by the Wilcoxon rank sum test, which is a non-parametric method, for the difference in the amount of CD9 protein expressed by the CD83 antibody. Wilcoxon rank sum test showed statistical significance when p value was less than 0.05.

As shown in FIG. 1, the microvessel was separated into anti-CD83 antibody (BD Pharmingen) and the amount of CD9 protein in the isolated microbesicles was confirmed by anti-CD9 antibody (R & D Systems) There was a statistically significant difference in the amount of microbicycles in the patient group. Based on these results, it was confirmed that the CD83 protein was expressed on the surface of the microbejac and 2) it was more concentrated on the microbeque of the breast cancer.

Example  2. Breast Cancer Related Microbesic  Micro RNA Marker  Selection

The plasma obtained in Example 1 The microvesicles were isolated with beads (Invitrogen) conjugated with anti-CD83 antibody (BD Pharmingen).

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed using Exiqon LNA ™ miRNA qRT-PCR panel (Exiqon).

The results of quantitative RT-PCR are shown in FIGS. 2A and 2B, and the results are shown in Table 1.

Surface protein of microbejac MicroRNA Amount of microRNA in benign tumor patients Amount of microRNA in breast cancer patients Difference in amount of microRNA
(? Cp) p-value
CD83 hsa-miR-126 30 26.3 -3.7 1.36E-08 hsa-miR-23a 30.6 27.5 -3.1 1.41E-04 hsa-miR-24 32 30.1 -1.9 2.98E-07 hsa-miR-19b 30.1 28.5 -1.6 8.61E-05 hsa-miR-103 34.2 31.9 -2.3 1.39E-04 hsa-miR-142-3p 30.8 28.7 -2.1 4.03E-05 hsa-miR-144 37.5 34.6 -2.9 8.23E-06 hsa-miR-15a 35.3 33.3 -2 1.20E-05 hsa-miR-185 36.1 34 -2.1 2.18E-05 hsa-miR-93 35.1 32.9 -2.2 1.65E-05 hsa-miR-30c 33.9 32 -1.9 6.01E-06

In FIGS. 2A, 2B, and 1, the smaller the Cp value is, the greater the expression is. The difference in the amount of microRNAs was analyzed by the Wilcoxon rank sum test. The Wilcoxon rank sum test showed statistical significance when the p-value was less than 0.05.

As shown in FIGS. 2A and 2B and Table 1, hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR- , hsa-miR-144, hsa-miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c were markers of breast cancer-related microvessels. The sequence of the miRNA is as follows.

hsa-miR-126; 5'-UCGUACCGUGAGUAAUAAUGCG-3 '(SEQ ID NO: 1)

hsa-miR-23a; 5'-AUCACAUUGCCAGGGAUUUCC-3 '(SEQ ID NO: 2)

hsa-miR-24; 5'-UGGCUCAGUUCAGCAGGAACAG-3 '(SEQ ID NO: 3)

hsa-miR-19b; 5'-UGUGCAAAUCCAUGCAAAACUGA-3 '(SEQ ID NO: 4)

hsa-miR-103; 5'-AGCAGCAUUGUACAGGGCUAUGA-3 '(SEQ ID NO: 5)

hsa-miR-142-3p; 5'-UGUAGUGUUUCCUACUUUAUGGA-3 '(SEQ ID NO: 6)

hsa-miR-144; 5'-UACAGUAUAGAUGAUGUACU-3 '(SEQ ID NO: 7)

hsa-miR-15a; 5'-UAGCAGCACAUAAUGGUUUGUG-3 '(SEQ ID NO: 8)

hsa-miR-185; 5'-UGGAGAGAAAGGCAGUUCCUGA-3 '(SEQ ID NO: 9)

hsa-miR-93; 5'-CAAAGUGCUGUUCGUGCAGGUAG-3 '(SEQ ID NO: 10)

hsa-miR-30c; 5'-UGUAAACAUCCUACACUCUCAGC-3 '(SEQ ID NO: 11)

Example  3. Breast Cancer Related Micro vizic  My Surface Protein and Micro RNA Evaluation of diagnostic accuracy

Predicted accuracy, area under the curve (AUC), and p-value were calculated for the diagnosis of breast cancer using the amount of surface protein and microRNA of microvessel. Samples obtained from 20 benign and 22 breast cancer patients were used. The accuracy of the prediction accuracy was defined as the agreement between actual breast cancer cases and predicted breast cancer cases. In addition, the AUC of 0.8 or more can be evaluated as having excellent effects.

Microvessel surface protein Types of Variables Number of variables Discriminant
(Logistic regression analysis) Prediction accuracy
(accuracy) AUC



CD83



Clinical information One age 0.62 0.62 Surface protein of microbejac One CD9 0.76 0.76 MiRNA 4 hsa-miR-126 +
hsa-miR-19b + hsa-miR-23a + hsa-miR-24 0.93 0.93 MicroRNA One hsa-miR-126 0.86 0.85 MicroRNA One hsa-miR-19b 0.71 0.71 MicroRNA One hsa-miR-23a 0.71 0.71 MicroRNA One hsa-miR-24 0.845 0.845 MicroRNA One hsa-miR-103 0.741 0.741 MicroRNA One hsa-miR-142-3p 0.741 0.741 MicroRNA One hsa-miR-144 0.69 0.69 MicroRNA One hsa-miR-15a 0.81 0.81 MicroRNA One hsa-miR-185 0.776 0.776 MicroRNA One hsa-miR-93 0.759 0.759 MicroRNA One hsa-miR-30c 0.776 0.776

As shown in Table 2, the amount of hsa-miR-126 contained in the microvasicles separated from the plasma by targeting CD83 can distinguish positive tumors and breast cancers with 86% accuracy, and predictive accuracy by hsa-miR-126 Was 24% higher than clinical information (age) and 10% higher than the amount of surface protein CD9.

<110> SAM SUNG ELECTRONICS <120> A composition and kit for diagnosing a breast cancer including a          polynucleotide within a vesicle, and method for diagnosing a          breast cancer using the same <130> PN098781 <160> 11 <170> Kopatentin 2.0 <210> 1 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-126 <400> 1 ucguaccgug aguaauaaug cg 22 <210> 2 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-23a <400> 2 aucacauugc cagggauuuc c 21 <210> 3 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-24 <400> 3 uggcucaguu cagcaggaac ag 22 <210> 4 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-19b <400> 4 ugugcaaauc caugcaaaac uga 23 <210> 5 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-103 <400> 5 agcagcauug uacagggcua uga 23 <210> 6 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-142-3p <400> 6 uguaguguuu ccuacuuuau gga 23 <210> 7 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-144 <400> 7 uacaguauag augauguacu 20 <210> 8 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-15a <400> 8 uagcagcaca uaaugguuug ug 22 <210> 9 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-185 <400> 9 uggagagaaa ggcaguuccu ga 22 <210> 10 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-93 <400> 10 caaagugcug uucgugcagg uag 23 <210> 11 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30c <400> 11 uguaaacauc cuacacucuc agc 23

Claims (23)

HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- one or more microRNAs (miRNA) selected from the group consisting of miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary to the fragments thereof / RTI &gt; The composition of claim 1, wherein the vesicle is a microbequique or exosome. The composition of claim 1, wherein the composition further comprises a substance capable of binding to a lipid bilayer of a substance or vesicle capable of specifically binding to the vesicle. The composition according to claim 3, wherein the substance capable of specifically binding to the vesicle is a surface protein, lipid, or a substance capable of binding to the sugar of the vesicle. The composition of claim 4, wherein the surface protein is CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2, . [Claim 4] The method according to claim 3, wherein the substance capable of specifically binding to the vesicle is selected from the group consisting of a substance having binding affinity to a protein, a substance capable of specifically binding to an enzyme substrate, a coenzyme, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, wherein the protein is selected from the group consisting of a protein, an antigen, an antibody or antigen binding fragment thereof, a hormone, &Lt; / RTI &gt; 4. The composition of claim 3, wherein the substance entrained in the lipid bilayer of the vesicle is a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. HSA-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa-miR- one or more microRNAs (miRNA) selected from the group consisting of miR-15a, hsa-miR-185, hsa-miR-93 and hsa-miR-30c, polynucleotides identical or complementary to the fragments thereof A breast cancer diagnostic kit. 9. The kit of claim 8, wherein the vesicle is a microbejac or exosome. 9. The kit of claim 8, wherein the kit further comprises a substance capable of binding to a lipid bilayer of a substance or vesicle capable of specifically binding to the vesicle. The kit according to claim 10, wherein the substance capable of specifically binding to the vesicle is a surface protein, lipid, or a substance capable of binding to the sugar of the vesicle. The kit according to claim 11, wherein the surface protein is CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2, . [Claim 10] The method according to claim 10, wherein the substance capable of specifically binding to the vesicle is selected from the group consisting of a substance having a binding affinity to a protein, a substance capable of specifically binding to an enzyme substrate, a coenzyme, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, in addition to an antibody, antigen, antibody or antigen binding fragment thereof, hormone, neurotransmitter, phospholipid- In kit. 11. The kit of claim 10, wherein the material interfering with the lipid bilayer of the vesicle is a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. Separating the vesicles from the biological sample separated from the subject;
Wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa miR-103, hsa-miR-142, hsa-miR-103, hsa-miR-142 and hsa-miR- Or more microRNA;
Comparing the amount of microRNAs measured with the amount of microRNAs measured from the vesicles isolated from the control sample; And
Determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample. 16. The method of claim 15, wherein the sample is at least one of urine, mucus, saliva, tears, blood, plasma, serum, sputum, spinal fluid, pleural effusion, aspiration nipple, lymphatic fluid, Cerebrospinal fluid, intracranial fluid, ascites, cystic tumor fluid, positive fluid, or a combination thereof. 16. The method of claim 15, wherein the vesicle is a microbequette or exosome. 16. The method of claim 15, wherein separating the vesicle from the biological sample comprises incubating the vesicle with a substance capable of interfering with the lipid bilayer of a substance or vesicle capable of specifically binding to the vesicle. 19. The method of claim 18, wherein the substance capable of specifically binding to the vesicle is a surface protein, lipid, or a substance capable of binding to the sugar of the vesicle. The method of claim 19, wherein the surface protein is CD83, CD9, EpCAM, caveolin, FasL, HLA-DRA, CD36, CD63, CD81, MUC1, ERBB4, GPER, ERBB2, MLANA, AMHR2, . [Claim 18] The method of claim 18, wherein the substance capable of specifically binding to the cell is selected from the group consisting of a substance having a binding affinity to a protein, a substrate of an enzyme, a coenzyme, a regulator, a substance specifically binding to a receptor, Binding protein, a protein comprising a pleckstrin homology (PH) domain, a cholesterol-binding protein, or a combination thereof, in addition to an antibody, antigen, antibody or antigen binding fragment thereof, hormone, neurotransmitter, phospholipid- / RTI &gt; 19. The method of claim 18, wherein the material having the property of interfering with the lipid bilayer of the vesicle is a material comprising a lipophilic moiety, an amphiphilic moiety, an amphoteric ion moiety, or a combination thereof. Separating the vesicles from the biological sample separated from the subject;
Wherein the microRNA is selected from the group consisting of hsa-miR-126, hsa-miR-23a, hsa-miR-24, hsa-miR-19b, hsa miR-103, hsa-miR-142, hsa-miR-103, hsa-miR-142 and hsa-miR- Or more microRNA;
Comparing the amount of microRNAs measured with the amount of microRNAs measured from the vesicles isolated from the control sample; And
Determining whether the amount of microRNAs measured is increased or decreased than the amount of microRNAs measured from the vesicles isolated from the control sample.

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