KR20180037462A - Composition for treatment and diagnosis of pancreatic neuroendocrine tumors - Google Patents

Abstract

The present invention relates to compositions and methods for diagnosis and prognosis of pancreatic neuroendocrine tumors. More particularly, the present invention relates to the use of HuD in the diagnosis and prognosis of pancreatic nerve endocrine tumors by confirming the association of pancreatic nerve endocrine tumors according to HuD expression For example.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for diagnosing pancreatic neuroendocrine tumors,

The present invention relates to a composition and method for diagnosis and prognosis of pancreatic neuroendocrine tumors, and more particularly, to a method for diagnosing pancreatic neuroendocrine tumors and predicting their prognosis by confirming the degree of expression of HuD.

The Hu family includes HuB, HuC and HuD, and these Elav / Hu proteins are known to bind to the U or AU-rich RNA sites of the target transcript through conserved RNA recognition motifs (RRMs 1-3) (Hinman and Lou, 2008; Pascale et al., 2008). Although HuD is known to promote neurite outgrowth (Kasashima et al., 1999; Abdelmohsen et al., 2010), the physiological role of HuD in animals is known to be complex. HuD targets a variety of proteins that are preferentially expressed in the nerve, such as GAP-43, acetylcholinesterase, tau, PSD-95, neuroserpin, and mRNA encoding musashi-1 do.

On the other hand, neuroendocrine tumors that occur mainly in the gastrointestinal tract, pancreas, and rectum are different in terms of symptoms and survival rates. Pancreatic neuroendocrine tumors (PNETs) are known to be a rare disease with a total prevalence of less than 1 per 100,000. The prognostic factors related to clinical features and survival were not known (Halfdanarson TR, Rabe KG, Rubin J, Petersen GM, Pancreatic neu-roendocrine tumors (PNETs): incidence, prognosis and recent trend toward improved survival. Ann Oncol 2008; 19: 1727-33), recent incidence is increasing (Fischer L, Bergmann F, Schimmack S, Hinzi, Priess S, Muller-Stich BP, et al. Outcome of surgery for pancreatic neuroendocrine neo-plasms. Br J Surg 2014; 101: 1405-12.). Pancreatic neuroendocrine tumors can be classified into functional and nonfunctional neuroendocrine tumors according to their clinical features. They are classified based on the clinical manifestations associated with active neuroendocrine hormones secreted by the tumor itself, which can not be distinguished microscopically . Although functional neuroendocrine tumors are known to have clinical symptoms associated with neuroendocrine hormones secreted from tumors, in clinical trials, even if they complain of such symptoms, it is difficult to diagnose them because of the rareness of the disease, There are many cases. In the case of functional neuroendocrine tumors of the pancreas, the incidence of malignant tumors with invasion or distant metastasis to the surrounding tissues was 10% for insulin, 60% for gastrin, 50% for nonfunctioning neuroendocrine tumors, (Pancreatic endocrine tumors: Diagnostic pitfalls. Hepa-togastroenterology 1999; 46: 679-90). The rate of pancreatic adenocarcinoma is slower than that of pancreatic adenocarcinoma. (Eriksson B, Oberg K. Neuroendocrine tumors of the pancreas. Br J Surg 2000; 87: 129-31).

The stage classification of pancreatic neuroendocrine tumors is largely divided into AJCC 7th edition and World Health Organization (WHO) classification. In the 7th edition of the American Joint Committee on Cancer (AJCC), pancreatic neuroendocrine tumors were introduced, but the classification of pancreatic cancer was applied. The WHO classification, which was revised in 2010, divided the tumors into three grades (Rindi G, Kloppel G, Alhman H, Caplin M, Virchows Arch 2006; 449: 395-401.) The tumor was found to be involved in the tumors that were included in the previous classification The size has been excluded and its clinical value has not yet been fully understood.

Pancreatic neuroendocrine tumors are not well known yet, but they are one of the most terrible diseases that lead to mortality due to poor prognosis when they occur in the pancreas, which is not well known by screening. In addition, since cancer has become more frequent in recent years, it is necessary to develop a diagnostic method based on research and interest of academia.

Rindi G, Kloppel G, Alhman H, Caplin M, Couvelard A, de Herder WW, et al. TNM staging of foregut (neuro) endocrine tumors: a consen- sus proposal including a grading system. Virchows Arch 2006; 449: 395-401.

In the present invention, the correlation between the amount of HuD expression and pancreatic endocrine neoplasia is investigated, and the present invention is applied to the diagnosis and prognosis of pancreatic endocrine tumors.

To achieve the above object, the present invention provides a composition for diagnosing pancreatic neuroendocrine tumors, comprising an agent for measuring the expression level of the HuD gene.

In addition, the present invention provides a composition for diagnosing pancreatic neuroendocrine tumor prognosis, comprising an antibody against HuD protein.

The present invention also provides a method for providing information for diagnosing pancreatic neuroendocrine tumors, comprising measuring the amount of HuD expression in a sample isolated from a human body.

In addition, the present invention provides a method for providing information for diagnosing a prognosis of a patient with pancreatic neuroendocrine tumor, comprising the step of measuring the amount of HuD expression in a sample separated from the human body.

According to the present invention, a decrease in HuD expression promotes pancreatic neuroendocrine tumor growth, and a decrease in HuD expression results in poor prognosis in patients with pancreatic neuroendocrine tumors. HuD overexpression inhibits pancreatic nerve endocrine tumors and HuD We conclude that HuD can be used for the diagnosis and prognosis of pancreatic nerve endocrine tumors because pancreatic endocrine tumor patients with increased expression have high survival rate and good prognosis.

FIG. 1 is a graph showing the degree of expression of HuD protein in tissues of a pancreas-derived neuroendocrine patient, and thus mitotic count and tumor size. FIG.
FIG. 2 shows the expression patterns of p27 and Ki-67 in HuD negative group and HuD positive group pancreas-derived neuroendocrine patient tissues.
FIG. 3 is a graph showing survival rate through Disease free survival (DFS) and progression free survival in human pancreas-derived neuroendocrine tumor patients according to HuD expression (HuD negative group and HuD positive group).
FIG. 4 is a graph showing the survival rate of an insulinoma cell line according to inhibition of HuD expression.
FIG. 5 is a graph showing cell cycle and cell growth of an insulinoma cell line according to inhibition of HuD expression.
Figure 6 shows the growth of insulinoma cell lines following inhibition of HuD expression Colony formation assay. ≪ / RTI >
FIG. 7 is a graph showing the insulin species formation through the size of insulin according to inhibition of HuD expression.
8 is a graph showing the survival rate of the insulinoma cell line overexpressing HuD.
FIG. 9 shows changes in insulin cell cycle and cell growth overexpressing HuD. FIG.
Figure 10 shows the growth of insulinoma cell lines with overexpression of HuD Colony formation assay.

The present invention will be described in detail. However, it should be understood that the present invention may be embodied with various changes and modifications without departing from the scope of the present invention.

The term "detection" or "measurement" as used herein means to quantify the concentration of a detected or measured subject.

As used herein, the phrase "agent for measuring the expression level of the HuD gene" means a molecule capable of detecting the mRNA level or the HuD protein level of the HuD gene by confirming the expression level of the HuD gene, Antibody, primer or probe.

As used herein, the term "primer" refers to a nucleic acid sequence having a short free 3 hydroxyl group that can form base pairs with a complementary template and is a starting point for template strand copy Short nucleic acid sequence " Primers can initiate DNA synthesis in the presence of reagents for the polymerization reaction (i. E., DNA polymerate or reverse transcriptase) and four different nucleoside triphosphates at the appropriate buffer solution and temperature. In the present invention, PCR amplification using a sense of HuD gene and an antisense primer can be used to determine the degree of HuD expression by measuring whether or not a desired product is produced and its level. The PCR conditions, the lengths of the sense and antisense primers can be modified on the basis of those known in the art, and thus the present invention is not particularly limited thereto.

In the present invention, the term "probe" means a nucleic acid fragment such as RNA or DNA corresponding to a short period of a few nucleotides or several hundreds of nucleotides capable of specifically binding with mRNA, . The probe may be prepared in the form of an oligonucleotide probe, a single stranded DNA probe, a double stranded DNA probe, or an RNA probe. In the present invention, hybridization using a probe complementary to the HuD gene can be used to diagnose the degree of HuD gene expression through hybridization. The selection and hybridization conditions of a suitable probe can be modified on the basis of those known in the art, so that the present invention is not particularly limited thereto.

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

The term "antibody" as used herein in the present invention means a specific protein molecule indicated for an antigenic site as a term known in the art. For the purpose of the present invention, the antibody refers to an antibody that specifically binds to a protein expressed in the HuD gene, which is a marker of the present invention, and the antibody can be prepared by a well-known method. Also included are partial peptides that can be made from the protein. The form of the antibody of the present invention is not particularly limited, and any polyclonal antibody, monoclonal antibody or antigen-binding antibody thereof may be included in the antibody of the present invention and include all immunoglobulin antibodies. Furthermore, the antibodies of the present invention include special antibodies such as humanized antibodies.

In one aspect, the present invention relates to a composition for diagnosing pancreatic neuroendocrine tumors, which comprises an agent for measuring the expression level of HuD (human antigen D) gene. In one embodiment, measuring the amount of expression of the HuD gene can be found by determining the level of expression of the HuD gene mRNA or the level of HuD protein encoded by the gene. RT-PCR, Competitive RT-PCR, Real-time RT-PCR, etc. can be used to identify the amount of HuD gene mRNA by using primer pairs or probes. , RNase protection assay (RPA), Northern blotting, DNA chip, and the like. The amount of the HuD protein encoded by the HuD gene can be determined by using an antibody that specifically binds to the protein. The analysis method includes Western blotting, enzyme linked immunosorbent assay (ELISA) Immunoassay, radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immuno staining, immunoprecipitation assay, complement fixation assay (RI) Complement Fixation Assay), FACS, protein chip, and the like. In one embodiment, the pancreatic neuroendocrine tumor may be a functional tumor associated with hormone secretion, may be an insulin species, and may be an insulinoma, a gastrin species or a glucagon species, depending on the type of hormone, but is most preferably an insulinoma.

In one aspect, the invention is directed to a composition for diagnosing pancreatic neuroendocrine tumor prognosis, comprising an antibody to a HuD protein. In one embodiment, the pancreatic neuroendocrine tumor may be a functional tumor associated with hormone secretion, may be an insulin species, and may be an insulinoma, a gastrin species or a glucagon species, depending on the type of hormone, but is most preferably an insulinoma.

In one aspect, the invention relates to a method of providing information for diagnosing pancreatic endocrine tumors, comprising measuring the amount of HuD expression in a sample isolated from the human body. In one embodiment, the pancreatic neuroendocrine tumor may be a functional tumor associated with hormone secretion, may be an insulin species, and may be an insulinoma, a gastrin species or a glucagon species, depending on the type of hormone, but is most preferably an insulinoma. In one embodiment, measurement of the amount of HuD expression can be confirmed by measuring the amount of HuD gene mRNA or HuD protein level. In one embodiment, the method comprises measuring the level of mRNA of the HuD gene or the expression level of the HuD gene in a sample isolated from a patient to provide information for pancreatic neuroendocrine tumor diagnosis; And comparing the level of expression of the measured mRNA or protein thereof with a control sample isolated from a patient who has not been afflicted with the disease, to provide information for diagnosis of pancreatic neuroendocrine tumors. In one embodiment, the method may include determining that the risk of developing pancreatic neuroendocrine tumors is high if the expression level is decreased by 50% or more compared to the amount of HuD expression of a normal human who has not been infected.

The sample of the patient includes, but is not limited to, tissue, cell, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid or urine. In addition, a specific method for measuring the expression level of the mRNA or the protein of the gene can detect the expression of the gene at the mRNA level or protein level, and the mRNA or protein is isolated from the biological sample using a known process can do. The mRNA or HuD protein levels of the HuD gene may be expressed as absolute (e.g., ug / ml) or relative (e.g., relative intensity of the signal) difference. Preferably, immunohistological staining using one or more antibodies against the Hud protein can be performed. After sampling and fixing the specimen from the patient, the paraffin-embedded block can be prepared by methods well known in the art. They are made into sections with a thickness of a few um and attached to a glass slide, and then the paraffin-embedded block can be reacted with any of the above-mentioned antibodies by a known method. The unreacted antibody may then be washed and labeled with one of the above-mentioned detection labels to read the label of the antibody on a microscope.

In one aspect, the invention relates to a method of providing information for the prognosis of a pancreatic neuroendocrine tumor patient, comprising measuring the amount of HuD expression in a sample isolated from the human body. In one embodiment, the prognosis of the pancreatic neuroendocrine tumor patient can be assessed for risk by Disease free survival (DFS) or progression free survival. In one embodiment, the amount of HuD expression can be determined by measuring the amount of HuD gene mRNA or HuD protein level. In one example, the HuD-positive group showed a significantly increased number of cells staying in the G1 phase compared to the negative group, and the mitotic cell was significantly decreased, and the cell division was markedly decreased (see Table 1). In one embodiment, the pancreatic neuroendocrine tumor may be a functional tumor associated with hormone secretion, may be an insulin species, and may be an insulinoma, a gastrin species or a glucagon species, depending on the type of hormone, but is most preferably an insulinoma. In one embodiment, the method may include determining that the risk of a prognosis is high if expression of the HuD is reduced by 50% or more relative to the amount of HuD expression in a normal human subject not afflicted with the disease.

The present invention will be described in more detail with reference to the following examples. However, the following examples are only for the purpose of illustrating the present invention, and thus the present invention is not limited thereto.

Example 1. Confirmation of correlation between HuD expression and pancreas-derived neuroendocrine tumors

1-1. Identification of HuD expression pattern in pancreas-derived neuroendocrine tumors

After staining 88 tissue of pancreas-derived neuroendocrine patients with anti-HuD antibody (Santa Cruz Biotechnology, Inc.), DAB signal intensity was analyzed by slide scanner and DAB signal intensity was analyzed to determine the degree of expression of HuD protein Respectively. The pancreatic tissues were classified into HuD negative group and HuD positive group according to the presence or absence of brightness of DAB signal specific to HuD protein. Clinical data of human pancreas-derived neuroendocrine tumors classified by lesion according to TNM (tumor-node-metastasis) classification were analyzed and the degree of expression of HuD (HuD negative group and HuD positive group) and tumor size, metastasis, And the results are shown in Table 1 below.

HuD Negative (23) Positive (65) P Sex male 43 13 (56.5) 20 (46.2) 0.393 female 45 10 (43.5) 35 (53.8) Age (year) 54 53.4 ± 13.5 52.3 ± 11.4 0.488 Size (cm) 2.4 4.6 ± 4.2 2.7 ± 2.8 0.006 * Grade by 2010 WHO classification G1 64 11 (47.8) 53 (81.5) <0.001 * G2 20 8 (34.8) 12 (18.5) G3 4 4 (17.4) 0 (0.0) Ki-67 index (%) 4.1 ± 8.9 1.2 ± 2.5 0.271 Mitotic count (10HPF) 5.9 ± 10.9 0.7 ± 2.1 0.001 * Extent of invasion Limited in pancreas 63 11 (47.8) 52 (80.0) 0.003 * Extended to other organ 25 12 (52.2) 13 (20.0) Angioinvasion Absent 58 11 (47.8) 47 (72.3) 0.033 * Present 30 12 (52.2) 18 (27.7) pT   ( AJCC  7 th  ed) One 32 3 (13.0) 29 (44.6) 0.004 * 2 31 8 (34.8) 23 (35.4) 3 25 12 (52.2) 13 (20.0) pN 0 80 21 (26.3) 59 (90.8) 0.980 One 8 2 (8.7) 6 (9.2) pM 0 81 21 (91.3) 60 (92.3) 1,000 One 7 2 (8.7) 5 (7.7) Associated syndrome Sporadic 80 18 (78.3) 62 (95.4) 0.009 * Von - Hippel - Lindau syndrom  ( VHL ) 5 2 (8.7) 3 (4.6) Multiple endocrine neoplasm (MEN) 3 3 (13.0) 0 (0.0)

mean SD, * P < 0.05

As a result, the mitotic count and tumor size of the HuD negative group were significantly increased compared to the HuD positive group, and the division ability of the tumor was increased (FIG. 1).

1-2. Confirm cell division through cell cycle analysis

In HuD negative group and HuD positive group, antibody specific for p27 protein, which is a cell cycle regulator, and Ki-67, a cell division marker (Santa Cruz Biotechnology, Inc.), was used for immunostaining Respectively. After dyeing, DAB signals were converted to digital images, and the differences between the two groups were quantitatively analyzed using Image J software.

As a result, the HuD negative group showed a decrease in p27 protein and an increase in Ki-67 compared to the HuD positive group (Fig. 2). In addition, we analyzed the correlation between expression of HuD and p27 using this, and confirmed that HuD and p27 were positively correlated.

Example  2. HuD  Expression and Pancreatic Origin Neuroendocrine tumor  Analysis of patient's prognosis

Clinical data of human pancreas-derived neuroendocrine tumors classified by lesion according to TNM (tumor-node-metastasis) classification were used to determine HuD expression, Disease free survival (DFS) and progression free survival ) Were analyzed using Excel and Graphpad.

As a result, the survival rate of the HuD negative group was significantly lower than that of the positive group, indicating that the patient with the HuD negative group showed a bad prognosis (FIG. 3). To further confirm the specificity of HuD protein in pancreatic neuroendocrine tumors, clinical indicators of pancreatic endocrine tumors were analyzed by univariate analysis and multivariate analysis according to presence or absence of HuD protein. As a result, it was confirmed that the relative expression level of HuD was significantly different from that of pancreatic endocrine neoplasm in both analyzes.

Univariate analysis Multivariate analysis N progress
n (%) Mean (month)
meanSE P hazard ratio
[95% CI] P HuD Negative 23 14 (60.9) 70 ± 11 <0.001 * 3.506 [1.246-9.871] 0.018 * Positive 65 9 (13.8) 181 ± 11 One Sex male 43 14 (32.6) 134 ± 18 0.042 * One female 45 9 (20.0) 114 ± 10 0.28 [0.095-0.827] 0.021 * Age (year) 0.998 [0.962-1.036] 0.931 Size () 1.298 [1.178-1.431] <0.001 * 1.209 [1.071-1.385] 0.002 * Grade by 2010 WHO classification G1 64 12 (18.7) 158 ± 17 0.001 * G2 20 7 (35.0) 88 ± 14 G3 4 4 (100.0) 38 ± 29 Ki-67 index (%) 1.042 [1.005-1.080] 0.025 * 1.01 [0.963-1.058] 0.694 Mitotic count (10HPF) 1.101 [1.059-1.145] <0.001 * Extent of invasion Limited in pancreas 63 10 (15.9) 148 ± 21 <0.001 * One Extended to other organ 25 13 (52.0) 70 ± 12 1.113 [0.332-3.726] 0.862 Angioinvasion Absent 58 7 (12.1) 170 ± 19 <0.001 * One Present 30 16 (53.3) 70 ± 11 3.338 [1.128-9.8722] 0.029 * Perineural invasion Absent 73 16 (21.9) 133 ± 18 0.005 * One Present 15 7 (46.7) 46 ± 8 0.842 [0.210-3.384] 0.843 pT (AJCC 7th ed) One 32 2 (6.2) 192 ± 16 <0.001 * 2 31 8 (25.8) 94 ± 11 3 25 13 (52.0) 70 ± 12 pN 0 80 19 (23.7) 129 ± 18 0.002 * One One 8 5 (62.5) 36 ± 11 3.097 0.680-14.119] 0.144 pM 0 81 18 (22.2) 128 0.003 * One One 7 5 (71.4) 59 2.029 [0.435-9.458] 0.368

by Cox regression analysis, hazard ratio [95% confidence interval], * P <0.05

Example  3. HuD  Through inhibition of expression Insulinoma  Growth confirmation

3-1. HuD  Suppression of expression Insulinoma  Determine cell viability

The mouse insulinoma cell line, βTC6, was cultured in DMEM media containing 10% FBS and 1% antibiotic. The HuD-specific siRNA (GCAUCCUGGUUGAUCAAGU) (Genolution Inc.) was injected into the cultured cell line in the form of liposome using lipofectamine 2000 (Invitrogen). Subsequently, 20mg / ml MTT solution was added to the cells, and after 16 hours, the survival rate of the cell line was confirmed by MTT assay comparing the degree of formation of formazane.

As a result, it was deduced that the inhibition of HuD increased the number of insulinoma cells by increasing the survival rate of the insulinoma cell line inhibiting the expression of HuD by siRNA (FIG. 4).

3-2. HuD  Suppression of expression Insulinoma  Identification of cell cycle and cell growth

As in Example 3-1, HuD siRNA was transfected into insulinoma cell line and cultured for 48 hours. The cultured cells were fixed in ethanol solution and analyzed for cell cycle changes using FACS equipment. In addition, control group treated with control siRNA and experimental group treated with HuD siRNA were inoculated into the same amount of culture dish, cultured for 3 days, and cell growth was analyzed by counting total cells using FACS equipment.

As a result, in HuD siRNA-treated cells, cells in the G2 / M phase of the cell cycle were increased, whereas cells in the G0G1 phase were decreased in comparison with the control group. Thus, the expression of HuD was decreased in the cells of the insulinoma cell line Increasing cleavage has been shown to promote cell growth and increase cell number (Figure 5).

3-3. HuD  Suppression of expression Insulinoma  Confirm cell growth

The control siRNA and HuD siRNA were each transfected into the insulinoma cell line as described in Example 3-1. The cells were then divided into 100 cells per 35 mm culture dish and cultured for 3 weeks to confirm the number of colonies formed.

As a result, it was found that insulinoma cells inhibiting HuD expression using siRNA significantly increased their growth compared to the control group (Fig. 6).

3-4. HuD  Suppression of expression Insulinoma  tumor Formability  Confirm

Diabetic model mice were injected with BALB / c at 6 weeks of age and a dose of 200 mg / Kg of streptozotocin. For transplantation of insulinoma cells, matrigel and insulinoma cells were mixed and injected three times a week into the subcutaneous tissues of mice. This procedure was performed 10 times in total. Tumor formation was observed for about 5 weeks after inoculation. All animal experiments were conducted in accordance with Ajou University and Catholic University Animal Experimental Research Ethics Code.

As a result, it was confirmed that in the case of insulinoma cells in which expression of HuD was inhibited, the tumor-forming ability was significantly increased as compared with the control cells without the expression of HuD (Fig. 7).

Example  4. HuD  Over-expression Insulinoma  growth Inhibition  Confirm

4-1. HuD  Overexpression Insulinoma  Determine cell viability

In contrast to Example 3-1., We examined the growth of insulin species by overexpression of HuD. Specifically, myc-tagged HuD plasmid was transfected into insulinoma cell line βTC6 using lipofectamine to induce overexpression of HuD, and cell viability of HuD-overexpressing cell line was confirmed by MTT assay.

As a result, the survival rate of HuD overexpressed insulinoma cell lines was reduced by more than 40%, suggesting that HuD induces the death of insulin species (FIG. 8).

4-2. HuD  Overexpression Insulinoma  Identification of cell cycle and cell growth

The myc-tagged HuD plasmid was transfected into the insulinoma cell line beta TC6 to overexpress HuD, and the cell cycle and cell growth were analyzed by the same method as in Example 4-2 using FACS equipment.

As a result, it was confirmed that, when HuD was overexpressed, cells of G2 / M group were decreased and cells of G0 / G1 group were increased compared to the control group, and that HuD overexpression inhibited the growth of insulinoma cells ).

4-3. HuD  Overexpression Insulinoma  Confirm cell growth

The myc-tagged HuD plasmid was transfected into the insulinoma cell line, beta TC6, and the cells were dispensed into a 35 mm culture dish of 100 cells each. Cell lines were cultured for 3 weeks after the dispensation and the number of colonies formed was confirmed.

As a result, the overexpression of HuD decreased the growth of the insulinoma cell line compared to the control (Fig. 10).

Claims (12)

A composition for diagnosing pancreatic nerve endocrine tumors, comprising an agent for measuring the expression level of a HuD gene. The composition for diagnosing pancreatic neuroendocrine tumors according to claim 1, wherein the measurement of the expression level of the HuD gene is a measurement of the mRNA level of HuD gene or the level of HuD protein. The composition for diagnosing pancreatic nerve endocrine tumors according to claim 1, wherein the pancreatic endocrine tumor is an insulin type. A composition for diagnosing pancreatic neuroendocrine tumor prognosis, comprising an antibody to a HuD protein. 5. The composition for diagnosing pancreatic neuroendocrine tumor according to claim 4, wherein the pancreatic endocrine tumor is an insulinoma. A method for providing information for diagnosing pancreatic neuroendocrine tumors, comprising measuring the amount of HuD expression in a sample separated from the human body. 7. The method of claim 6, wherein the pancreatic endocrine tumor is an insulinoma. 7. The method according to claim 6, wherein the measurement of the amount of HuD expression is performed by measuring the amount of HuD gene mRNA or HuD protein level. A method for providing information for diagnosing a prognosis of a pancreatic neuroendocrine tumor patient, comprising the step of measuring the amount of HuD expression in a sample isolated from a human body. The method according to claim 9, wherein the prognosis of the pancreatic neuroendocrine tumor patient is DFS (Diseasefree survival) or progression free survival. . 10. The method according to claim 9, wherein the measurement of the amount of HuD expression is performed by measuring the amount of mRNA of the HuD gene or the level of HuD protein to confirm the prognosis of the pancreatic endocrine tumor patient. 10. The method of claim 9, wherein the pancreatic endocrine tumor is an insulinoma.

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