KR20210052348A - Composition for Differential Diagnosis of Thyroid Follicular Neoplasm and Differential Diagnosis Method Using the Same - Google Patents

Abstract

The present invention relates to a composition for differential diagnosis of thyroid follicular tumor, a kit for differential diagnosis comprising the same, and a method for providing information necessary for differential diagnosis of thyroid follicular tumor using the same. According to the present invention, thyroid follicular cancer can be differentiated from other benign thyroid tumors, such as thyroid hyperproliferative nodules or thyroid follicular adenomas, and diagnosed without surgery.

Description

Composition for Differential Diagnosis of Thyroid Follicular Neoplasm and Differential Diagnosis Method Using the Same}

The present invention relates to a composition for differential diagnosis of thyroid follicular tumor, a differential diagnosis kit including the same, and a method of providing information necessary for differential diagnosis of thyroid follicular tumor using the same.

Follicular thyroid carcinoma (FTC) accounts for about 20% of all thyroid cancers and belongs to differentiated thyroid cancer with a good prognosis. Thyroid follicular cancer is mainly present at an older age compared to papillary thyroid carcinoma, which accounts for the majority of thyroid cancer, whereas thyroid follicular cancer metastases mainly through lymph nodes, thyroid follicular cancer metastases through blood vessels There is a difference. On the other hand, thyroid papillary cancer can be diagnosed relatively easily by observing the characteristic nucleus morphology with a fine needle aspiration test. However, thyroid follicular cancer is a benign tumor of follicular adenoma (FA) depending on whether the specimen has encapsulated after surgery. Because they are distinguished, it is impossible to diagnose by microneedle aspiration test or central needle biopsy before surgery.

Therefore, currently, the only method to confirm thyroid follicular cancer is to perform a thyroidectomy through surgery and then collect tissue to confirm the pathology within the tissue. In this pathology test including surgery, first half of the thyroid gland in which nodules were found is surgically resected and a biopsy is performed. Afterwards, if thyroid follicular cancer is diagnosed, except for the early cancer with a good prognosis, the other half through surgery is performed again. This is a very cumbersome method because it has to be removed again. Therefore, there is a need to develop a biomarker capable of distinguishing between thyroid follicular cancer and follicular adenoma and confirming thyroid follicular cancer in thyroid tissue or blood obtained during fine needle aspiration test or central needle biopsy. In particular, when a protein-based biomarker is developed, information necessary for diagnosis can be quickly derived by measuring the expression level of the corresponding marker using an antibody or the like, and there is no need to perform surgery for pathology examination.

Among the mutations of genes such as BRAF, RET/PTC, RAS, PAX8/PPAR gamma, and P53, which have been proposed as biomarkers of thyroid follicular cancer, RAS mutations, which have a relatively high incidence rate, are frequently found in follicular adenoma. Because the ratio is not high, it is not actually used for diagnosis. In addition, a study confirming that the expression levels of five genes, such as ELMO1, EMCN, ITIH5, KCNAB1, and SLCO2A1, decreased in thyroid follicular cancer tissue compared to follicular adenoma tissue, the possibility of being developed as a marker that can distinguish between thyroid follicular cancer and follicular adenoma. However, it has many limitations in its use as a marker, such as having to go through the process of extracting mRNA from tissues. Protein markers that discriminate thyroid cancer are being used as immunostaining methods (immunohistochemistry), but their specificity is so low that there is little possibility of using them to distinguish between thyroid follicular cancer and follicular adenoma.

Accordingly, there is an urgent need to develop a biomarker that can clearly distinguish thyroid follicular cancer from benign tumors such as thyroid hyperproliferative nodules or precancerous lesions such as follicular adenoma.

Aleksandra Pfeifer et al. Molecular differential diagnosis of follicular thyroid carcinoma and adenoma based on gene expression profiling by using formalin-fixed paraffin-embedded tissues. BMC medical genomics, vol. 6:38. 7 Oct. 2013, doi:10.1186/1755-8794-6-38

Accordingly, the present inventors performed extensive quantitative and qualitative proteomic analysis using follicular thyroid cancer (FTC), nodular hyperplasia (NH) and thyroid follicular adenoma (FA) tissues. The present invention was completed by discovering a protein specifically expressed in thyroid follicular cancer tissue.

Accordingly, an object of the present invention is to provide a composition for differential diagnosis of thyroid follicular neoplasm, comprising, as an active ingredient, an agent for measuring the expression level of one or more proteins selected from the group consisting of proteins described in Table 2 below or mRNA expression level thereof. To provide.

[Table 2]

Another object of the present invention is to provide a kit for differential diagnosis of thyroid follicular tumor, comprising the composition according to the present invention.

Another object of the present invention is required for differential diagnosis of thyroid follicular tumor, comprising the step of measuring the expression level of one or more proteins selected from the group consisting of the proteins listed in Table 2 or mRNA expression levels in a biological sample isolated from a subject. It is to provide a way to provide information.

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention, the present invention provides a composition for differential diagnosis of thyroid follicle tumor, comprising as an active ingredient one or more proteins selected from the group consisting of proteins described in Table 2 below or an agent for measuring the mRNA expression level thereof. to provide:

[Table 2]

In one embodiment of the present invention, the composition may further include an agent for measuring the mRNA expression level of one or more proteins selected from the group consisting of proteins shown in Table 3 below, but is not limited thereto:

[Table 3]

In another embodiment of the present invention, the thyroid follicular tumor may be one or more selected from the group consisting of thyroid follicular cancer, thyroid follicular adenoma, and thyroid hyperproliferative nodule.

In another embodiment of the present invention, the composition may diagnose thyroid follicular cancer by distinguishing it from thyroid follicular adenoma or thyroid hyperproliferative nodule.

In another embodiment of the present invention, the agent for measuring the expression level of the mRNA may be a primer set or probe that specifically binds to the mRNA, but is not limited thereto.

In another embodiment of the present invention, the agent for measuring the expression level of the protein may be an antibody or aptamer specific for the protein, but is not limited thereto.

In another embodiment of the present invention, the composition may be used to determine whether or not a thyroid follicular tumor is operated and a range of surgery, but is not limited thereto.

In addition, the present invention provides a kit for differential diagnosis of thyroid follicular tumor comprising the composition according to the present invention.

In addition, the present invention provides information necessary for differential diagnosis of thyroid follicular tumor, comprising measuring the expression level of one or more proteins selected from the group consisting of the proteins listed in Table 2 or mRNA expression levels in a biological sample isolated from a subject. Provides a way to provide.

In one embodiment of the present invention, the method may further include measuring the expression level of one or more proteins selected from the group consisting of the proteins shown in Table 3 or mRNA expression levels thereof, but is not limited thereto.

In another embodiment of the present invention, the method may further include determining as thyroid follicular cancer when the expression of one or more proteins selected from the group consisting of the proteins listed in Table 2 or mRNA thereof is detected or increased. However, it is not limited thereto.

In another embodiment of the present invention, the subject may be a patient suspected of having a thyroid follicular tumor, but is not limited thereto.

In another embodiment of the present invention, the biological sample is blood, whole blood, plasma, urine, tissue, cells, organs, bone marrow, microneedle aspiration sample, microneedle washing fluid, core needle biopsy sample and saliva. It may be one or more selected from the group consisting of, but is not limited thereto.

In another embodiment of the present invention, the protein expression level is Western blot, ELISA, radioimmunoassay, radioimmunoassay, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunostaining, It may be measured by one or more methods selected from the group consisting of immunoprecipitation assay, complement fixation assay, mass spectrometry, FACS, and protein chip, but is not limited thereto.

In another embodiment of the present invention, the mRNA expression level is at least one selected from the group consisting of PCR, RNase protection assay, northern blotting, southern blotting, in situ hybridization, and DNA chip. It can be measured by a method, but is not limited thereto.

In addition, the present invention provides a use of an agent for measuring the expression level of one or more proteins selected from the group consisting of the proteins listed in Table 2 or mRNA expression levels thereof, for differential diagnosis of thyroid follicular tumors.

In addition, the present invention provides a method for differential diagnosis of thyroid follicular tumor, comprising the step of measuring the expression level of one or more proteins selected from the group consisting of the proteins described in Table 2 or mRNA thereof.

The present invention relates to a composition for differential diagnosis of thyroid follicular tumor and a method for differential diagnosis of thyroid follicular tumor using the same, and establishing a method for diagnosing thyroid follicular cancer before surgery using the composition capable of detecting a protein marker according to the present invention By doing so, there is an effect of being able to distinguish and diagnose thyroid follicular cancer from other benign thyroid tumors such as thyroid hyperproliferative nodules or thyroid follicular adenoma without surgery. Accordingly, it is expected that unnecessary surgery and patient pain can be reduced.

1 shows a time series flow chart of quantitative proteomic analysis using thyroid FFPE (Formalin-fixed, Paraffin-embedded) tissue.
Figure 2a shows a peptide spectrum abundance distribution for thyroid follicular cancer, thyroid hyperproliferative nodules, and thyroid follicular adenoma tissue samples.
Figure 2b shows the results of PCA (principal component analysis) (FA, thyroid follicular adenoma; FTC, thyroid follicular cancer; NH, thyroid hyperproliferative nodule).
Figure 3a shows the results of qualitative proteomic analysis for three groups FTC, FA, and NH using Venn diagram analysis.
3b and 3c show quantitative proteomic analysis results for three groups FTC, FA, and NH using a volcano plot analysis.FIG. 3B is an analysis of FTC/NH, and FIG. 3C is an analysis of FA/NH. will be.
4 is a diagram showing a flow chart of a thyroid follicular cancer (FTC) specific protein screening process.

The present invention is a composition for differential diagnosis of thyroid follicular neoplasm comprising, as an active ingredient, an agent for measuring the expression level of one or more proteins selected from the group consisting of proteins described in Table 2 below or mRNA expression level thereof, and a differential diagnosis method using the same Etc.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for differential diagnosis of thyroid follicular tumor, comprising as an active ingredient an agent for measuring the expression level of one or more proteins selected from the group consisting of proteins described in Table 2 below or mRNA expression level thereof:

[Table 2]

In the present invention, the composition may further include an agent for measuring the mRNA expression level of one or more proteins selected from the group consisting of the proteins shown in Table 3 below:

[Table 3]

As used herein, the term “thyroid follicular neoplasm” refers to any follicular neoplasm that may occur in the thyroid gland, and is used interchangeably with the term “follicular thyroid neoplasm”. I can. The “thyroid follicular tumor” is a clinical diagnosis preliminarily given through a thyroid examination before surgery, and includes all benign adenoma, nodular hyperplasia, and malignant carcinoma throughout the present specification. Was used as.

As used herein, the term “protein” is used interchangeably with “polypeptide” or “peptide”, for example, refers to a polymer of amino acid residues as commonly found in proteins in their natural state. .

As used herein, the term “expression” means that a protein or nucleic acid is produced in a cell.

As used herein, the term “protein” is used interchangeably with “polypeptide” or “peptide”, for example, refers to a polymer of amino acid residues as commonly found in proteins in their natural state. . “Polynucleotide” or “nucleic acid” refers to deoxyribonucleotides (DNA) or ribonucleotides (RNA) in the form of single- or double-stranded. Unless otherwise limited, known analogues of natural nucleotides that hybridize to nucleic acids in a manner similar to those of naturally occurring nucleotides are also included. “MRNA” refers to RNA that transfers genetic information (gene-specific nucleotide sequence) to ribosomes that specify amino acid sequences from a specific gene in the process of protein synthesis.

As used herein, the term “differential diagnosis” refers to distinguishing a particular disease or condition from others exhibiting similar symptoms. The differential diagnosis method is a systematic diagnosis method used to confirm the existence of a state in which multiple alternatives are possible. This method is essentially a removal process or an information acquisition process that reduces the probability of a candidate state to a negligible level. The term is also used to refer to determining whether a thyroid follicular tumor is a thyroid follicular cancer, a benign follicular tumor-for example a follicular adenoma or a hyperproliferative nodule.

In the present invention, the thyroid follicular tumor may be one or more selected from the group consisting of thyroid follicular cancer, thyroid follicular adenoma, and thyroid hyperproliferative nodules.

The term "follicular thyroid carcinoma (FTC)" as used herein is the second most common thyroid cancer after papillary thyroid carcinoma (PTC), which occurs in about 10%, and has a high incidence in iodine deficient areas. There is a tendency. Although follicular cancer is classified as a well-differentiated thyroid cancer, such as papillary cancer, it develops at an older age and shows other clinical pathological characteristics, such as a more aggressive tendency than papillary cancer. Follicular cancer is a malignant tumor arising from the epithelium of the thyroid gland, and it is possible to diagnose histologically based on the presence of infiltration and distant metastasis into the capsule, lymphatic vessels, blood vessels or surrounding tissues, unlike papillary cancer. FNA) or a frozen section test during surgery is almost impossible to make an accurate diagnosis. Follicular cancer, follicular adenoma, etc. can be diagnosed as follicular neoplasm by a preoperative cell aspiration test. Follicular cancer is usually a minimally invasive type that involves localized encapsulation or vascular invasion within the tumor, or both, and a broadly invasive type characterized by gross encapsulation or invasion into the surrounding tissues of the thyroid gland ( widely invasive type). Among these, there are reports that microinvasive follicular cancers have different prognosis depending on film or vascular invasion, and microinvasive follicular cancers with vascular invasion are further classified as moderately invasive, regardless of the encapsulation. have. However, there is no method to diagnose thyroid follicular cancer before surgery, so if it is clinically suspected, it is diagnosed through surgery, but only 10-40% of cases are confirmed as follicular cancer in diagnostic surgery. Moreover, many of the patients who have not been diagnosed with follicular cancer after surgery require supplementation with thyroid hormones for life after unnecessary surgery, and patients diagnosed with follicular cancer after surgery have additional residual thyroidectomy and isotopes depending on the stage/recurrence risk. The disadvantage is that treatment is required. Various immunostaining and genetic analysis have been applied to preoperative diagnosis, but there is no clearly proven method until now, and according to recent studies, it is impossible to distinguish between malignant and benign even with a change in size.

As mentioned above, "papillary thyroid cancer" is the most common type of thyroid cancer, accounting for about 70% of all thyroid cancers, and occurs in middle-aged women. It is known that it responds relatively well to treatment and has a good prognosis. Papillary cancer generally grows very slowly and has a good prognosis. Papillary cancer may occur only in one lobe of the thyroid gland, but in 20 to 45% of all papillary cancers, both lobes are invaded (multiple), and spread to lymph nodes (lymph glands) around the thyroid gland is also observed in as many as 40%. do. In this case, if appropriate treatment is received early, most of them heal well, but in rare cases, there are cases of distant metastasis to other parts such as lungs or bones, so early detection and treatment are important. In the case of papillary cancer, even if cancer is found in only one lobe of the thyroid gland, when encapsulation or lymph node metastasis is suspected on preoperative ultrasound, total thyroidectomy is performed to remove the entire thyroid gland, and the cancer often invades the central cervical lymph node. Therefore, when performing a total thyroidectomy, the thyroid gland and the central cervical lymph node are usually removed together.

The composition of the present invention can diagnose thyroid follicular cancer by distinguishing it from thyroid follicular adenoma or thyroid hyperproliferative nodule.

When the composition of the present invention is for measuring the expression level of mRNA, the agent for measuring the expression level of mRNA may be a primer set or probe that specifically binds to the mRNA.

The composition of the present invention comprising a primer set or probe specific for the mRNA of the protein described in Table 2 or Table 3 may further include an agent required for a known method of detecting RNA. By using a known method for detecting RNA using the composition of the present invention without limitation, the level of mRNA of the protein markers in a subject can be measured.

As used herein, the term “primer” refers to a short nucleic acid sequence that can form a base pair with a complementary template as a nucleic acid sequence having a short free three-terminal hydroxyl group and serves as a starting point for template strand copying. . Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates at an appropriate buffer and temperature.

As used herein, the term “probe” refers to a nucleic acid fragment such as RNA or DNA corresponding to a few bases or hundreds of bases that can achieve specific binding to mRNA, and is labeled so that a specific mRNA You can check the presence or absence of. The probe may be manufactured in the form of an oligonucleotide probe, a single stranded DNA probe, a double stranded DNA probe, an RNA probe, or the like. Selection of suitable probes and conditions for hybridization can be modified based on those known in the art.

The “primer” or “probe” can be chemically synthesized using a method for synthesizing a phosphoramidite solid support or other well-known methods. In addition, primers or probes can be variously modified according to methods known in the art within a range that does not interfere with hybridization with mRNA. Examples of such modifications include methylation, encapsulation, substitution of one or more homologs of natural nucleotides, and modifications between nucleotides, such as uncharged linkers (e.g. methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.). ) Or charged linkers (eg, phosphorothioate, phosphorodithioate, etc.), and binding of a labeling material using fluorescence or enzymes.

Meanwhile, the agent for measuring the expression level of the protein in the present invention may be an antibody or aptamer that specifically binds to the protein.

As used herein, the term “antibody” refers to a specific protein molecule directed against an antigenic site. For the purposes of the present invention, an antibody refers to an antibody that specifically binds to a marker protein, and includes all of polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. In addition, as long as it has antigen-antibody binding, a part of the whole antibody is also included in the antibody of the present invention, and all kinds of immunoglobulin antibodies that specifically bind to the protein of the present invention are included. For example, a complete antibody having two full-length light chains and two full-length heavy chains, as well as functional fragments of antibody molecules, i.e., Fab, F(ab'), F(ab') with antigen-binding function 2 and Fv, and the like. Further, the antibody of the present invention includes special antibodies such as humanized antibodies and chimeric antibodies, and recombinant antibodies as long as it can specifically bind to the protein of the present invention.

As used herein, the term “aptamer” is a substance capable of specifically binding to an analyte to be detected in a sample, and a single-stranded nucleic acid (DNA, RNA, or modified Nucleic acid), and specifically, the presence of a target protein in a sample can be confirmed. The production of aptamer is synthesized by determining the sequence of an oligonucleotide having a high binding ability and selective to the target protein to be identified according to a general production method of aptamer, and then the 5'or 3'end of the oligonucleotide In order to bind to the functional group of the timer chip, it may be _{made by modifying it with -SH, -COOH, -OH or -NH 2} , but is not limited thereto.

The composition of the present invention comprising an antibody specific to the protein described in Table 2 or Table 3 may additionally contain an agent required for a method for detecting a known protein, and a method for detecting a known protein using the present composition Without limitation, the expression level of the protein in the subject (in the present invention, a biological sample obtained from the subject) can be measured.

In the present invention, since the nucleic acid sequence of the gene encoding the protein marker is registered in the gene bank, those skilled in the art design antisense oligonucleotides, primer pairs, or probes that specifically amplify specific regions of these genes based on the sequence. can do.

The antisense oligonucleotide can be chemically synthesized using a phosphoramidite solid support method, or other well-known methods, and includes all functional equivalents. In addition, the above-described “primer” or “probe” may be applied in the same manner.

The term "functional equivalent" refers to, for example, one or more substitutions, deletions or additions from the reference sequence, the actual effect that does not result in various functional dissimilarities between the reference sequence and the subject sequence. It refers to both nucleotide and nucleic acid sequences of mutant sequences that have been changed, such as (net effect). Typically, such a substantially equivalent sequence is only about 35% (i.e., the number of substitutions, additions, and deletions of each residue in the substantially equivalent sequence is compared to the corresponding reference sequence and the remaining total number in the substantially equivalent sequence. Divided by about 0.35 or less), it varies from those listed in this specification. Such sequences have 65% sequence identity with the listed sequence. Substantially equivalents according to the invention, for example mutant, amino acid sequences have preferably at least 80% sequence identity, more preferably at least 90% sequence identity with the listed amino acid sequence. Substantially equivalent nucleotide sequences of the invention may have a lower percentage of sequence identity, for example, taking into account the redundancy or degeneracy of the genetic code. Preferably, the nucleotide sequence should have at least about 65% identity, more preferably at least about 75% identity, and most preferably about 95% identity. For the purposes of the present invention, sequences having substantially equivalent biological activity and substantially equivalent synthetic characteristics are treated as substantial equivalents. In order to determine the equivalent, the truncation of the mature sequence (eg, through a mutation producing a spurious stop codon) should be ignored.

In the present invention, the composition may be used to determine whether or not a thyroid follicle tumor is operated and a surgical range, but is not limited thereto.

As another aspect of the present invention, the present invention can provide a kit for differential diagnosis of thyroid follicular tumor comprising the composition according to the present invention.

The kit of the present invention may include an antibody that recognizes a target protein as a marker or a primer set that recognizes an mRNA as a marker, a probe, as well as one or more other constituent compositions, solutions, or devices suitable for an analysis method.

As a specific embodiment, the diagnostic kit may be a diagnostic kit comprising essential elements necessary to perform a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit contains each set of primers specific for a marker gene. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of each marker gene, and is about 7 bp to 50 bp in length, more preferably about 10 bp to 30 bp in length. In addition, a primer specific to the nucleic acid sequence of the control gene may be included. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (various in pH and magnesium concentration), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNAse, RNAse inhibitors DEPC. -May include DEPC-water, sterilized water, etc.

In another specific aspect, it may be a diagnostic kit comprising essential elements necessary to perform a DNA chip. The DNA chip kit may include a substrate to which cDNA or oligonucleotide corresponding to a gene or fragment thereof is attached, and reagents, agents, enzymes, etc. for preparing a fluorescently labeled probe. In addition, the substrate may include a cDNA or oligonucleotide corresponding to a control gene or a fragment thereof.

In another aspect of the present invention, the present invention is characterized by the step of measuring the mRNA expression level of one or more proteins selected from the group consisting of the proteins listed in Table 2 in a biological sample isolated from a subject or differentiation of thyroid follicular tumors. It can provide a method of providing information necessary for diagnosis.

In the present invention, the method may further include measuring the expression level of one or more proteins selected from the group consisting of the proteins shown in Table 3 or mRNA expression levels thereof, but is not limited thereto.

In the present invention, the method may further include determining as thyroid follicular cancer when the expression of one or more proteins selected from the group consisting of the proteins listed in Table 2 or mRNA thereof is detected or increased.

As used herein, the term “detection” is to measure and confirm the presence (expression) of a target substance (marker protein in the present invention), or to measure a change in the abundance level (expression level) of a target substance. And it means to include all things to confirm. In the same context, measuring the expression level of the protein refers to measuring the expression or not (ie, measuring the presence or absence of expression), or measuring the level of qualitative or quantitative change of the protein. The measurement may be performed without limitation, including both a qualitative method (analysis) and a quantitative method. The types of qualitative and quantitative methods for measuring protein levels are well known in the art, and the experimental methods described herein are included therein. Specific protein level comparison methods for each method are well known in the art. Therefore, the detection of the target protein is meant to include detection of the presence or absence of the protein shown in Table 2 or Table 3, or confirming an increase (up-regulation) or decrease (down-regulation) of the protein expression level.

In addition, the term “diagnosis” as used herein refers to determining the susceptibility of a subject to a specific disease or disease, determining whether a subject currently has a specific disease or disease, or a specific disease. Or determining the prognosis of the diseased subject, or therametrics (eg, monitoring the condition of the subject to provide information on treatment efficacy).

In the present invention, the subject may be a patient suspected of having a thyroid follicular tumor, but is not limited thereto. For example, the subject may be a patient suspected of a thyroid tumor, and the subject in the method according to the present invention was preliminarily diagnosed as a follicular tumor, but a patient suspected of all thyroid follicular cancer or thyroid follicular adenoma Can be In other words, the subject is a patient who has a tumor or nodule in the thyroid gland and needs to distinguish whether the tumor or nodule is malignant thyroid follicular cancer, a benign thyroid hyperproliferative nodule or follicular adenoma. Such patients include those who initially developed a thyroid tumor and those who were previously diagnosed with a thyroid tumor.

In the present invention, the biological sample may be used without limitation as long as it is collected from a subject for differential diagnosis of a thyroid tumor, and for example, tissue, cells, blood, plasma, serum, saliva, etc. obtained by biopsy, etc. It includes not only biological samples such as nasal fluid, sputum, joint capsule fluid, amniotic fluid, ascites, cervical secretion, vaginal secretion, urine, cerebrospinal fluid, etc., as well as microneedle aspiration samples and microneedle wash solutions. Preferably, in the group consisting of biological liquid samples, such as blood, whole blood, plasma, urine, tissues, cells, organs, bone marrow, microneedle aspiration samples, microneedle wash solutions, core needle biopsy samples, and saliva. It can be measured in one or more selected. The present invention can be diagnosed through a body fluid sample (e.g., plasma) other than a thyroid tumor or a direct thyroid lesion where a nodule has occurred, and thus the effectiveness, rapidity, convenience, and safety of diagnosis are increased.

The biological sample can be pretreated prior to use for detection or diagnosis. For example, it may include homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, and the like. The sample may be prepared to increase the detection sensitivity of the protein marker. For example, a serum sample obtained from a patient is anion exchange chromatography, affinity chromatography, size exclusion chromatography, liquid chromatography, It can be pretreated using a method such as sequential extraction or gel electrophoresis.

As used herein, the term “fine needle aspiration cytology (FNAC)” refers to a benign or malignant thyroid tumor or a nodule that is suspected of being observed with a microscopic needle to obtain a small amount of cells while viewing ultrasound. It is performed for the purpose of diagnosing the cells of the patient, and is a criterion for determining the progression of thyroid surgery, the extent of surgery, and the extent of lymph node resection by aspirating cells by stabbing the nodule with a thin injection needle.

The measurement of the protein expression level is not particularly limited as long as it is by a protein expression measurement method known in the art.For example, Western blot, ELISA, radioimmunoassay, radioimmune diffusion method, Ouchterlony immune diffusion method (Ouchterlony immunodiffusion), rocket immunoelectrophoresis, immunostaining, immunoprecipitation assay, complement fixation assay, mass spectrometry, FACS, and protein chip.

The gene expression level measurement is a group consisting of PCR, RNase protection assay, northern blotting, southern blotting, in situ hybridization, and DNA chip according to a gene expression measurement method known in the art. It may be one or more selected from.

As used herein, the term'analysis' may preferably mean'measurement', and the qualitative analysis may mean measuring and confirming the presence or absence of a target substance, and the quantitative analysis It may mean measuring and confirming a change in the presence level (expression level) or amount of a target substance. In the present invention, the analysis or measurement may be performed without limitation, including both a qualitative method and a quantitative method, and preferably, a quantitative measurement may be performed.

Terms used in the present invention have selected general terms that are currently widely used as possible while taking functions of the present invention into consideration, but this may vary according to the intention or precedent of a technician working in the field, the emergence of new technologies, and the like. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning of the terms will be described in detail in the description of the corresponding invention. Therefore, the terms used in the present invention should be defined based on the meaning of the term and the overall contents of the present invention, not a simple name of the term.

Throughout the specification of the present invention, when a certain part “includes” a certain constituent element, it means that other constituent elements may be further included rather than excluding other constituent elements unless otherwise stated. The terms "about", "substantially", etc. of the degree used throughout the specification of the present invention are used at or close to the numerical value when manufacturing and material tolerances specific to the stated meaning are presented, and the present invention To aid in understanding of, accurate or absolute figures are used to prevent unreasonable use of the stated disclosure by unscrupulous infringers.

Throughout the specification of the present invention, the term "combination of these" included in the expression of the Makushi format refers to one or more mixtures or combinations selected from the group consisting of the constituent elements described in the expression of the Makushi format. It means to include at least one selected from the group consisting of constituent elements.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the examples.

[Example]

Preparation for experiment

1. Sample preparation

Thyroid tumor tissue was obtained from patients with thyroid follicular cancer, thyroid follicular adenoma, and thyroid hyperproliferative nodule, respectively, and proteomic analysis was performed using paraffinized tissue after surgery.

As shown in the flowchart of FIG. 1, the thyroid FFPE (Formalin-fixed, Paraffin-embedded) tissue was removed from paraffin using heptane/methanol, and proteins were extracted. Thereafter, precipitation was performed using acetone, and the protein was quantified by a quantitative analysis of BCA (bicinchoninic acid) protein.

300 μg of protein was denatured and reduced using 8M urea to which 250mM TCEP was added, followed by alkylation with 55mM IAA. Then, the protein was digested with LysC containing trypsin, and desalted using C18 reverse phase chromatography.

The sample obtained as described above was dried and analyzed by LC-HRMS.

2. LC-MS analysis

Mass spectrometry (hereinafter referred to as'MS') equipment is Q-Exactive Plus BioPharm version (ThermoFisher, USA), and liquid chromatography (hereinafter referred to as'LC') is UltiMate™3000 RSLCnano System (ThermoFisher). , USA) was used. Peptide samples were injected 5ul each and analyzed with an ion trap mass spectrometer linked to NanoLC at a flow rate of 350nl/min.

The column used for liquid chromatography was a fused silica capillary column C18 having a length of 50 cm, an inner diameter of 75 μm, and an outer diameter of 360 μm. Samples were analyzed at a flow rate of 360 nl/min for 200 minutes, and the concentration gradient was from 5% Solution A (0.1% formic acid w/ 5% DMSO) to 50% Solution B (80% acetonitrile, 0.1% formic acid w/ 5%). DMSO) for 150 minutes.

The separated peptide was ion-sprayed using an electrospray ionization voltage of 2000V, and the ion emitter was injected into a mass spectrometer through a 20um inner diameter tappered ion spray emitter to obtain spectral data.

The parameters for mass spectrometry were MS1 resolution 70000, MS1 maximum fill time 20msec, DDA (data dependent acquisition) method (top 20), MS2 resolution 17500, MS2 maximum fill time 100msec, AGC (auto gain control). ) Carried out with 1e6.

Example 1. Analysis of proteomic bodies in thyroid follicular cancer, thyroid follicular adenoma, and thyroid hyperproliferative nodule tissue

Proteomic analysis was performed on thyroid hyperproliferative nodules (NH), thyroid follicular adenoma (FA), and thyroid follicular cancer (FTC) tissue samples (a total of 30 pieces of 10 pieces each). As a result, as shown in FIG. 2A, distribution of peptide intensity within the spectrum obtained from each sample was analyzed reproducibly for all 30 samples.

In addition, a principal component analysis (PCA) was performed using quantitative information on the proteomics of each of the 30 samples. As a result, as shown in Fig. 2b, each of the ten tissues of the thyroid hyperproliferative nodule, thyroid follicular cancer, and thyroid follicular adenoma was found to be well distinguished.

Example 2. Proteomic analysis and derivation of thyroid follicular cancer-specific protein through it

2.1. Qualitative protein analysis using Venn diagram analysis

As a result of qualitative analysis of the protein using a Venn diagram, as shown in the graph (Size of each list) in FIG. 3A, 10 thyroid hyperproliferative nodules (NH, indicated in green in FIG. 3A) tissues contained 3,579 proteins, and 10 thyroid glands. In follicular cancer (FTC, shown in blue in FIG. 3A) tissue, 3,867 proteins were identified, and in 10 thyroid follicular adenoma tissues (FA, indicated in red in FIG. 3A), 3,672 proteins were identified.

In addition, as shown in the Venn diagram of FIG. 3A, a total of 174 thyroid follicular cancer-specific proteins, which are biomarkers of the present invention, were identified, and a total of 34 thyroid follicular adenoma-specific proteins were identified.

Although not identified in thyroid hyperproliferative nodules, a total of 147 proteins were commonly identified in thyroid follicular cancer and thyroid follicular adenoma.

2.2. Quantitative proteomic analysis using Volcano plot analysis

As a result of quantitative analysis of the protein through volcano plot analysis, as shown in FIG. 3b, a total of 432 proteins were identified as having a P value of 0.05 or less in the thyroid follicular cancer group compared to the thyroid hyperproliferative nodule group, increasing more than two times, and 2 Proteins that decrease by less than a fold were identified as 78 proteins.

In addition, as shown in Figure 3c, compared with the thyroid hyperproliferative nodule group, the P value in the thyroid follicular adenoma group was found to be 0.05 or less, and the number of proteins that increased more than two times was confirmed to be a total of 331, and the number of proteins that decreased by 2 times or less was 203. Confirmed.

2.3. Derivation of thyroid follicular cancer-specific protein

In summary, the thyroid follicular cancer-specific proteins that were not detected in the thyroid follicular adenoma group and the thyroid hyperproliferative nodule group are shown in Table 1 below. When using the proteins shown in Table 1 below, since it is possible to diagnose thyroid follicular cancer even before surgery, it can be utilized as a biomarker for differential diagnosis of thyroid follicular cancer.

Thyroid follicular cancer (FTC) specific protein not detected in thyroid hyperproliferative nodules (NH) and thyroid follicular adenoma (FA) (primary screening) Accession No. Thyroid follicular cancer specific protein (174) Q9BV57 1,2-dihydroxy-3-keto-5-methylthiopentene dioxygenase O15235 28S ribosomal protein S12, mitochondrial P62851 40S ribosomal protein S25 Q9NZ32 Actin-related protein 10 Q8N3C0 Activating signal cointegrator 1 complex subunit 3 Q8WXI4 Acyl-coenzyme A thioesterase 11 Q9ULX6 A-kinase anchor protein 8-like Q13424 Alpha-1-syntrophin Q01484 Ankyrin-2 P61966 AP-1 complex subunit sigma-1A Q9NVP1 ATP-dependent RNA helicase DDX18 Q9NR09 Baculoviral IAP repeat-containing protein 6 O15155 BET1 homolog P78537 Biogenesis of lysosome-related organelles complex 1 subunit 1 Q9NP80 Calcium-independent phospholipase A2-gamma P49674 Casein kinase I isoform epsilon Q9H078 Caseinolytic peptidase B protein homolog Q8WXE0 Caskin-2 P26992 Ciliary neurotrophic factor receptor subunit alpha O15551 Claudin-3 P00742 Coagulation factor X P02452 Collagen alpha-1(I) chain Q969H4 Connector enhancer of kinase suppressor of ras 1 Q8IWV2 Contactin-4 Q92793 CREB-binding protein Q7KZN9 Cytochrome c oxidase assembly protein COX15 homolog P00395 Cytochrome c oxidase subunit 1 P00414 Cytochrome c oxidase subunit 3 Q9H0P0 Cytosolic 5'-nucleotidase 3A Q9Y4B6 DDB1- and CUL4-associated factor 1 Q86YH6 Decaprenyl-diphosphate synthase subunit 2 Q5JSL3 Dedicator of cytokinesis protein 11 Q9H6A0 DENN domain-containing protein 2D Q9UI17 Dimethylglycine dehydrogenase, mitochondrial Q01459 Di-N-acetylchitobiase Q9Y485 DmX-like protein 1 P49736 DNA replication licensing factor MCM2 P33993 DNA replication licensing factor MCM7 Q96LL9 DnaJ homolog subfamily C member 30 Q15075 Early endosome antigen 1 O14638 Ectonucleotide pyrophosphatase/phosphodiesterase family member 3 O94919 Endonuclease domain-containing 1 protein Q96RQ1 Endoplasmic reticulum-Golgi intermediate compartment protein 2 Q7L8L6 FAST kinase domain-containing protein 5, mitochondrial Q9Y613 FH1/FH2 domain-containing protein 1 Q2TAA5 GDP-Man:Man(3)GlcNAc(2)-PP-Dol alpha-1,2-mannosyltransferase Q8WUA4 General transcription factor 3C polypeptide 2 Q9NWU2 Glucose-induced degradation protein 8 homolog Q16769 Glutaminyl-peptide cyclotransferase Q8WWP7 GTPase IMAP family member 1 Q8ND71 GTPase IMAP family member 8 P63096 Guanine nucleotide-binding protein G(i) subunit alpha-1 P48723 Heat shock 70 kDa protein 13 O95757 Heat shock 70 kDa protein 4L Q9Y241 HIG1 domain family member 1A, mitochondrial P0C0S5 Histone H2A.Z P01892 HLA class I histocompatibility antigen, A-2 alpha chain P01920 HLA class II histocompatibility antigen, DQ beta 1 chain O00629 Importin subunit alpha-3 Q5JVS0 Intracellular hyaluronan-binding protein 4 Q9NR19 Isoform 2 of Acetyl-coenzyme A synthetase, cytoplasmic Q9Y2D5 Isoform 2 of A-kinase anchor protein 2 O15446 Isoform 2 of DNA-directed RNA polymerase I subunit RPA34 Q9H0Q3 Isoform 2 of FXYD domain-containing ion transport regulator 6 Q14353 Isoform 2 of Guanidinoacetate N-methyltransferase Q7Z4I7 Isoform 2 of LIM and senescent cell antigen-like-containing domain protein 2 O43148 Isoform 2 of mRNA cap guanine-N7 methyltransferase Q13356 Isoform 2 of Peptidyl-prolyl cis-trans isomerase-like 2 Q14CB8 Isoform 2 of Rho GTPase-activating protein 19 Q9BPX6 Isoform 3 of Calcium uptake protein 1, mitochondrial Q8NEU8 Isoform 3 of DCC-interacting protein 13-beta Q7L099 Isoform 3 of Protein RUFY3 O95294 Isoform 3 of RasGAP-activating-like protein 1 Q15424 Isoform 3 of Scaffold attachment factor B1 Q9NRL3 Isoform 3 of Striatin-4 Q9NW81 Isoform 4 of ATP synthase subunit s-like protein Q9Y2J2 Isoform 4 of Band 4.1-like protein 3 O75909 Isoform 4 of Cyclin-K Q9BQS8 Isoform 4 of FYVE and coiled-coil domain-containing protein 1 Q14168 Isoform 4 of MAGUK p55 subfamily member 2 P46087 Isoform 4 of Probable 28S rRNA (cytosine(4447)-C(5))-methyltransferase Q96P11 Isoform 4 of Probable 28S rRNA (cytosine-C(5))-methyltransferase Q5ST30 Isoform 4 of Valine--tRNA ligase, mitochondrial P49750 Isoform 4 of YLP motif-containing protein 1 Q7Z6K5 Isoform C15orf38-AP3S2 of Arpin P09496 Isoform Non-brain of Clathrin light chain A Q8IXV7 Kelch domain-containing protein 8B Q8NBF6 Late secretory pathway protein AVL9 homolog Q15334 Lethal(2) giant larvae protein homolog 1 Q9UQ13 Leucine-rich repeat protein SHOC-2 Q9NZU1 Leucine-rich repeat transmembrane protein FLRT1 Q9NR99 Matrix-remodeling-associated protein 5 Q687X5 Metalloreductase STEAP4 Q9BTC8 Metastasis-associated protein MTA3 Q9BVV7 Mitochondrial import inner membrane translocase subunit Tim21 Q9Y5J7 Mitochondrial import inner membrane translocase subunit Tim9 Q969M1 Mitochondrial import receptor subunit TOM40B Q9H8L6 Multimerin-2 Q02083 N-acylethanolamine-hydrolyzing acid amidase P03905 NADH-ubiquinone oxidoreductase chain 4 P22307 Non-specific lipid-transfer protein Q9BVI4 Nucleolar complex protein 4 homolog Q9BZE4 Nucleolar GTP-binding protein 1 P78316 Nucleolar protein 14 Q9NR30 Nucleolar RNA helicase 2 P31483 Nucleolysin TIA-1 isoform p40 Q9H4L5 Oxysterol-binding protein-related protein 3 Q9C0B5 Palmitoyltransferase ZDHHC5 Q9Y3E5 Peptidyl-tRNA hydrolase 2, mitochondrial Q96BW9 Phosphatidate cytidylyltransferase, mitochondrial O15305 Phosphomannomutase 2 Q9HAB8 Phosphopantothenate--cysteine ligase Q96JA3 Pleckstrin homology domain-containing family A member 8 Q6UN15 Pre-mRNA 3'-end-processing factor FIP1 O60508 Pre-mRNA-processing factor 17 Q86Y79 Probable peptidyl-tRNA hydrolase P98196 Probable phospholipid-transporting ATPase IH O95456 Proteasome assembly chaperone 1 Q96DE9 Protein CXorf40B Q96C01 Protein FAM136A Q9BQD7 Protein FAM173A Q96ND0 Protein FAM210A Q969G5 Protein kinase C delta-binding protein B7ZBB8 Protein phosphatase 1 regulatory subunit 3G A6NK58 Putative lipoyltransferase 2, mitochondrial Q5SRD1 Putative mitochondrial import inner membrane translocase subunit Tim23B Q9HBR0 Putative sodium-coupled neutral amino acid transporter 10 Q96GD0 Pyridoxal phosphate phosphatase Q53H96 Pyrroline-5-carboxylate reductase 3 P04626 receptor tyrosine-protein kinase erbB-2 O75787 Renin receptor Q9NWS8 Required for meiotic nuclear division protein 1 homolog Q9UKL0 REST corepressor 1 Q12774 Rho guanine nucleotide exchange factor 5 Q9NP77 RNA polymerase II subunit A C-terminal domain phosphatase SSU72 Q70HW3 S-adenosylmethionine mitochondrial carrier protein Q08AF3 Schlafen family member 5 P62341 Selenoprotein T Q9NRX5 Serine incorporator 1 Q13523 Serine/threonine-protein kinase PRP4 homolog Q13315 Serine-protein kinase ATM Q9H788 SH2 domain-containing protein 4A Q9UJC5 SH3 domain-binding glutamic acid-rich-like protein 2 Q9Y6A9 Signal peptidase complex subunit 1 Q8NCG7 Sn1-specific diacylglycerol lipase beta P19634 Sodium/hydrogen exchanger 1 P11166 Solute carrier family 2, facilitated glucose transporter member 1 O15020 Spectrin beta chain, non-erythrocytic 2 P11474 Steroid hormone receptor ERR1 Q8WU79 Stromal membrane-associated protein 2 Q96JG6 Syndetin O14662 Syntaxin-16 Q5T5C0 Syntaxin-binding protein 5 Q15554 Telomeric repeat-binding factor 2 Q53QW1 Testis-expressed protein 44 Q9UL33 Trafficking protein particle complex subunit 2-like protein Q00403 Transcription initiation factor IIB P01137 Transforming growth factor beta-1 Q8WUH2 Transforming growth factor-beta receptor-associated protein 1 Q14232 Translation initiation factor eIF-2B subunit alpha O15321 Transmembrane 9 superfamily member 1 Q9HC07 Transmembrane protein 165 Q9H649 tRNA (cytosine(34)-C(5))-methyltransferase, mitochondrial Q32P41 tRNA (guanine(37)-N1)-methyltransferase Q70EL2 Ubiquitin carboxyl-terminal hydrolase 45 Q96K76 Ubiquitin carboxyl-terminal hydrolase 47 Q9H8M7 Ubiquitin carboxyl-terminal hydrolase MINDY-3 O14562 Ubiquitin domain-containing protein UBFD1 Q96N11 Uncharacterized protein C7orf26 O00160 Unconventional myosin-If Q9H267 Vacuolar protein sorting-associated protein 33B Q9H898 Zinc finger matrin-type protein 4 Q9Y6M5 Zinc transporter 1 Q15043 Zinc transporter ZIP14

As described above, in order to differentially diagnose thyroid follicular cancer from thyroid follicular tumor, the protein(s) disclosed in Table 1 may be used.

Example 3. Selection of final marker protein for differential diagnosis of thyroid follicular cancer

Of the marker proteins listed in Table 1, 13 protein markers were finally selected in consideration of experimental significance, detection possibility in various specimens, and ease of detection.

Specifically, according to Examples 1 and 2, proteins that were not detected in the thyroid follicular adenoma group and the thyroid hyperproliferative nodule group but were identified with high significance only in the thyroid follicular cancer group were first selected. Among the first-selected proteins, proteins that satisfy all of the following four criteria were secondarily selected:

① Proteins registered in the plasma proteome database;

② proteins registered in the exosome proteome database;

③ The scaled abundance measured in LS-MS data is 50 or higher; And

④ Peptide spectrum matched (PSM) of 2 or more in LS-MS data.

Criteria ① and ② above are not only in tumor cells obtained by tissue, microneedle aspiration, but also in relatively specimens by additionally selecting proteins present in plasma, blood, tumor tissue, or exosomes secreted from cells among the first selected protein markers. It is to differentially diagnose thyroid follicular cancer using the protein marker according to the present invention in plasma, blood, whole blood, urine, microneedle wash, saliva, and exosomes contained therein.

In addition, the criteria ③ and ④ are intended to facilitate detection because they exist relatively more abundantly among the first-selected protein markers, and the number of matching peptide spectra is at least 2 and thus easy to discover.

Protein markers finally selected through the first and second selection processes are shown in Table 2 below.

Thyroid follicular cancer (FTC)-specific protein not detected in thyroid hyperproliferative nodules (NH) and thyroid follicular adenoma (FA) (secondary screening) Accession No. gene ID Thyroid follicular cancer specific protein (13) PPD ^One) EPD ²⁾ SA ³⁾ CV% #P ⁴⁾ PSM ⁶⁾ P0C0S5 H2AFZ Histone H2A.Z HPRD_00823 O 300 64.32 4 385 O95757 HSPA4L Heat shock 70 kDa protein 4L HPRD_07137 O 300 81.77 5 12 Q9BTC8 MTA3 Metastasis-associated protein MTA3 HPRD_12359 O 85.4 31.91 2 7 Q13523 PRPF4B Serine/threonine-protein kinase PRP4 homolog HPRD_09087 O 300 45.78 2 5 P02452 COL1A1 Collagen alpha-1(I) chain HPRD_00362 O 300 73.02 One 4 P62851 RPS25 40S ribosomal protein S25 HPRD_01593 O 300 156.11 2 4 Q15043 SLC39A14 Zinc transporter ZIP14 HPRD_12288 O 300 62.41 One 3 P11166 SLC2A1 Solute carrier family 2, facilitated glucose transporter member 1 HPRD_00683 O 300 72.47 One 3 Q9NR09 BIRC6 Baculoviral IAP repeat-containing protein 6 HPRD_05731 O 300 26.6 One 2 Q16769 QPCT Glutaminyl-peptide cyclotransferase HPRD_06140 O 300 45.75 2 2 Q687X5 STEAP4 Metalloreductase STEAP4 HPRD_11637 O 300 48.16 2 2 Q8N3C0 ASCC3 Activating signal cointegrator 1 complex subunit 3 HPRD_09980 O 300 54.11 One 2 P26992 CNTFR Ciliary neurotrophic factor receptor subunit alpha HPRD_00348 O 300 64.57 2 2

^One) PPD, Plasma Proteome Database; ²⁾ EPD, Exosome Proteome Database; ³⁾ SA, Scaled Abundance; ⁴⁾ #P, Number of Peptide; ⁵⁾ PSM, Peptide Spectrum Matched

In summary, the protein marker of the present invention can quickly and easily differentiate and diagnose thyroid follicular cancer from thyroid hyperproliferative nodules or thyroid follicular adenoma even before surgery for follicular tumors in the thyroid gland. By accurately diagnosing thyroid follicular cancer, which was impossible, it is expected that unnecessary surgery can be reduced and only thyroid follicular cancer that requires surgery can be treated.

In addition, the composition according to the present invention is produced as a kit, and is expected to be used to quickly examine whether a thyroid tumor is undergoing surgery from a patient sample.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that other specific forms can be easily modified without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and are not limiting.

Claims (15)

A composition for differential diagnosis of thyroid follicular tumor, comprising as an active ingredient an agent for measuring the expression level of one or more proteins selected from the group consisting of proteins described in Table 2 below or mRNA expression level thereof:
[Table 2]

The method of claim 1,
The composition is a composition for differential diagnosis of thyroid follicular tumor, characterized in that it further comprises an agent for measuring the expression level of one or more proteins selected from the group consisting of the proteins shown in Table 3 or mRNA thereof:
[Table 3]

The method of claim 1,
The thyroid follicular tumor is a composition for differential diagnosis of thyroid follicular tumor, characterized in that at least one selected from the group consisting of thyroid follicular cancer, thyroid follicular adenoma, and thyroid hyperproliferative nodule.
The method of claim 1,
The composition is a composition for differential diagnosis of thyroid follicular tumor, characterized in that the thyroid follicular cancer can be diagnosed by distinguishing it from thyroid follicular adenoma or thyroid hyperproliferative nodule.
The method of claim 1,
The agent for measuring the expression level of the mRNA is characterized in that the primer set or probe that specifically binds to the mRNA, a composition for differential diagnosis of thyroid follicular tumor.
The method of claim 1,
The composition for differential diagnosis of thyroid follicular tumor, characterized in that the agent for measuring the expression level of the protein is an antibody or aptamer specific for the protein.
The method of claim 1,
The composition is for determining whether or not a thyroid follicle tumor is operated and a surgical range, a composition for differential diagnosis of a thyroid follicle tumor.
A kit for differential diagnosis of thyroid follicular tumor, comprising the composition of any one of claims 1 to 7.
A method of providing information necessary for differential diagnosis of thyroid follicular tumor, comprising measuring the mRNA expression level of one or more proteins selected from the group consisting of the proteins shown in Table 2 below in a biological sample isolated from a subject:
[Table 2]

The method of claim 9,
The method further comprises the step of measuring the expression level of one or more proteins selected from the group consisting of the proteins listed in Table 3 below, or mRNA expression level thereof, a method of providing information necessary for differential diagnosis of thyroid follicular tumors:
[Table 3]

The method of claim 9,
The method further comprises the step of determining as thyroid follicular cancer when the expression of one or more proteins selected from the group consisting of the proteins described in Table 2 or mRNA thereof is detected or increased. How to provide the necessary information for differential diagnosis.
The method of claim 9,
The method for providing information necessary for differential diagnosis of thyroid follicular tumor, characterized in that the subject is a patient suspected of having a thyroid follicular tumor.
The method of claim 9,
The biological sample is at least one selected from the group consisting of blood, whole blood, plasma, urine, tissue, cells, organs, bone marrow, microneedle aspiration samples, microneedle wash solutions, core needle biopsy samples, and saliva. A method of providing information necessary for differential diagnosis of thyroid follicular tumor.
The method of claim 9,
The protein expression level was Western blot, ELISA, radioimmunoassay, radioimmunoassay, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunostaining, immunoprecipitation assay, complement fixation assay, mass Method for providing information necessary for differential diagnosis of thyroid follicular tumor, characterized in that it is measured by one or more methods selected from the group consisting of mass spectrometry, FACS, and protein chips.
The method of claim 9,
The mRNA expression level is characterized in that it is measured by one or more methods selected from the group consisting of PCR, RNase protection assay, northern blotting, southern blotting, in situ hybridization, and DNA chip. A method of providing information necessary for differential diagnosis of thyroid follicular tumors.

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