METTL7A methylation site and application thereof

Abstract

The invention relates to application of the expression level or methylation level of METTL7A in diagnosis of silent pituitary neuroendocrine tumors. The methylation level of the METTL7A comprises the methylation level of a methylation site cg05097930 of the METTL7A. The application can be used for preoperative diagnosis of subtypes of silent pituitary neuroendocrine tumors, solves the hysteresis and limitation of dependence on postoperative pathology and iconography in the prior art, and realizes early precise diagnosis and treatment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of molecular diagnosis, in particular to METTL7A methylation site and application thereof, more particularly to application of METTL7A methylation level in distinguishing silent adrenocorticotropic hormone pituitary neuroendocrine tumor and silent gonadotropin pituitary neuroendocrine tumor. BACKGROUND

[0002] Pituitary neuroendocrine tumor (PitNET) is one of the most common intracranial tumors, accounting for about 15% of all intracranial tumors. A series of clinical manifestations of PitNET are caused by excessive hormone secretion and invasion of surrounding structures (such as II-V cranial nerves, hypothalamus and internal carotid artery). Although most PitNETs are considered to be benign, more than 40% of PitNETs require invasive surgical treatment. Due to the difficulty of complete surgical resection and the limitation of radiotherapy and chemotherapy options, the treatment of PitNETs is still challenging.

[0003] Classification of PitNETs is based on immunohistochemistry and transcription factor expression, such as PIT-1, TPIT, and SF-1 lineage. Silent PitNETs can be divided into 8 subtypes according to immunohistochemical expression of pituitary hormones and pituitary-specific transcription factors: silent gonadotroph adenomas (SGAs), silent corticotroph adenomas (SCAs), silent growth hormone adenomas (SGHA), silent TSH secreting adenomas (STAs), silent prolactinomas (SPAs), multiple hormone PIT-1 positive pituitary neuroendocrine tumors, zero cell hormone pituitary neuroendocrine tumors, dual / triple pituitary neuroendocrine tumors. Among the 8 subtypes of silent PitNETs, silent corticotroph adenomas (SCAs) and gonadotroph adenomas (SGAs) are the most common. SCAs are one of the most common silent adenomas, accounting for 3%-19% of non-functioning adenomas. SCAs belong to the corticotroph lineage, and immunohistochemistry shows ACTH positivity, pituitary transcription factor is T-PIT, and there is no clinical manifestation of increased cortisol secretion and Cushing's disease. Compared with other silent pituitary endocrine tumors, SCAs have more common sphenoid sinus invasion and higher recurrence rate in patients. SGAs are also one of the most common silent adenomas, most of which are gonadotroph tumors. Gonadotroph adenomas are usually silent tumors and only in a few cases can cause clinical syndromes related to gonadotrophin secretion. The recurrence rate of silent gonadotroph pituitary neuroendocrine tumors is lower than that of silent prolactin pituitary neuroendocrine tumors, silent growth hormone pituitary neuroendocrine tumors, and silent corticotroph pituitary neuroendocrine tumors. However, the identification of SCAs and SGAs still mainly relies on conventional methods such as postoperative pathology.

[0004] Aggressive pituitary tumours (APTs): tumours that are unresponsive to standard treatment (i.e. surgery, conventional medical treatment and radiotherapy) and exhibit multiple local recurrences. The abnormal rapid growth and / or recurrence of aggressive pituitary neuroendocrine tumours can lead to an increase in morbidity and mortality in patients. The mechanism of pituitary neuroendocrine tumour invasiveness is not yet clear. At present, the invasiveness of tumours is mainly diagnosed by conventional methods such as imaging (tumour growth invading the cavernous sinus), postoperative pathology (high mitotic index, ki67 index > 3%) and the like, which have their limitations.

[0005] According to the WHO 2022 classification, the main method for identifying the invasiveness of silent pituitary neuroendocrine tumour (PITNETs) subtypes is as follows: 1. Transcription factor-based molecular typing based on PIT1, TPIT and SF1 can distinguish silent subtypes (such as silent ACTH tumour). In TPIT tumours, splice diversity (especially ESRP1-regulated splicing events) is associated with invasiveness. Patients in the high PSI (splicing acceptance rate) group have a poorer prognosis, and ESRP1 dysregulation is an important marker. 2. Gene mutation: Gsα (gsp) mutation is found in 40% of invasive GH tumours; PKCα mutation promotes local infiltration by activating collagenase; high expression of PTTG induces angiogenesis and infiltration. 3. Pathological and clinical integrated assessment: according to the WHO 2022 classification, the invasiveness is determined by combining Ki-67 index, mitotic count and tumour infiltration (such as cavernous sinus invasion). Combined with intraoperative features: hard texture of the tumour, wrapping around blood vessels or tight adhesion suggest invasiveness, but still need to be confirmed by postoperative pathology. Surgical resection is the main treatment, and it is difficult to individualize the preoperative diagnosis and treatment plan to reduce the recurrence rate of patients (3.9 years of follow-up, 29% of SCA patients need to undergo a second surgery). Therefore, the index for preoperative differential diagnosis of pituitary neuroendocrine tumour subtypes and invasiveness is urgently needed. The research on plasma biomarkers for the differential diagnosis of silent pituitary neuroendocrine tumours (silent PITNETs) is still in the exploratory stage, only a few molecular markers show potential value, and new biomarkers are urgently needed.

[0006] The classic oncogenes and tumor suppressor genes are absent in pituitary tumors, so more and more researchers begin to study the changes of DNA promoter methylation. Studies have shown that genes involved in cell growth and signal transduction, such as RAS family members, pituitary tumor apoptosis gene (PTAG), etc., all show changes in methylation status. The increase of DNA methyltransferase (DNMT1 and DNMT3A) may also be related to the invasiveness of pituitary tumors. Studies have shown that chemokines (CCL2, CCL5, etc.) regulate tumor invasion and metastasis through DNA methylation of molecules such as STAT3. Plasma ctDNA (circulating tumor DNA) can capture tumor-specific gene mutations to assist diagnosis. The level of ctDNA is related to tumor burden and treatment response. Plasma ctDNA methylation is closely related to the invasiveness of pituitary neuroendocrine tumors, but no study has screened out related DNA methylation indicators in the plasma of patients with high sensitivity and consistent with tissue expression. SUMMARY

[0007] The purpose of the present application is to provide a detection agent for obtaining the expression level or methylation level of METTL7A in a sample in the preparation of a diagnostic product for silent pituitary neuroendocrine tumors, or the application of the expression level or methylation level of METTL7A in the diagnosis of silent pituitary neuroendocrine tumors.

[0008] In one aspect, the present application provides a detection agent for obtaining the expression level and / or methylation level of METTL7A (methyltransferase like 7A) in a sample in the preparation of a diagnostic product for silent pituitary neuroendocrine tumors.

[0009] In one embodiment, the methylation level of METTL7A is the methylation level of METTL7A methylation site cg05097930.

[0010] In one embodiment, the diagnosis includes early screening.

[0011] In one embodiment, the detection agent is one or several detection agents used in methylation-specific PCR (MSP), methylation sequencing, methylation chip or pyrosequencing.

[0012] In one embodiment, the sample is a plasma sample and / or a tumor tissue sample.

[0013] In one embodiment, the silent pituitary neuroendocrine tumor comprises silent gonadotroph adenomas (SGAs), silent corticotroph adenomas (SCAs), silent growth hormone adenomas (SGHA), silent TSH secreting adenomas (STAs), silent prolactinomas (SPAs), multiple hormone PIT-1 positive pituitary neuroendocrine tumors, zero cell hormone pituitary neuroendocrine tumors, and dual / triple pituitary neuroendocrine tumors.

[0014] In one embodiment, the silent pituitary neuroendocrine tumor comprises silent corticotroph adenomas and silent gonadotroph adenomas.

[0015] In one embodiment, the silent pituitary neuroendocrine tumor is silent corticotroph adenomas and silent gonadotroph adenomas.

[0016] In one embodiment, the GENEBANK ID of the METTL7A is 25840.

[0017] In one embodiment, the methylation level of the METTL7A comprises the methylation level of the METTL7A methylation site cg05097930.

[0018] In one embodiment, the methylation level of the METTL7A is the methylation level of the METTL7A methylation site cg05097930.

[0019] Preferably, the use comprises use of a reagent for detecting the expression level or methylation level of METTL7A in the preparation of a kit for differentiating silent corticotroph adenomas and silent gonadotroph adenomas.

[0020] Preferably, the use comprises use of a reagent for detecting the methylation level of the METTL7A methylation site cg05097930 in the preparation of a kit for differentiating silent corticotroph adenomas and silent gonadotroph adenomas.

[0021] In one embodiment, the detection agent or reagent comprises a methylation detection primer pair, the sequence of which is shown as SEQ ID No. 1-2.

[0022] In another aspect, the present application provides use of METTL7A expression level and / or methylation level in diagnosis of silent pituitary neuroendocrine tumors.

[0023] In another aspect, the present application provides use of METTL7A methylation site as a diagnostic marker for silent pituitary neuroendocrine tumors; preferably, the METTL7A methylation site is METTL7A gene cg05097930 methylation site.

[0024] In another aspect, the present application provides a product for diagnosis of silent pituitary neuroendocrine tumors, wherein the product comprises reagents for detecting METTL7A expression level or methylation level.

[0025] In one embodiment, the product is in the form of a kit.

[0026] In another aspect, the present application provides use of a reagent for detecting METTL7A expression level and / or methylation level in preparation of a kit for identifying subtypes of silent pituitary neuroendocrine tumors.

[0027] In one embodiment, the subtypes of silent pituitary neuroendocrine tumors comprise silent gonadotroph adenomas (SGAs), silent corticotroph adenomas (SCAs), silent growth hormone adenomas (SGHA), silent TSH secreting adenomas (STAs), silent prolactinomas (SPAs), multiple hormone PIT-1 positive pituitary neuroendocrine tumors, zero cell hormone pituitary neuroendocrine tumors, and dual / triple pituitary neuroendocrine tumors.

[0028] In one embodiment, the subtypes of silent pituitary neuroendocrine tumors comprise silent adrenocorticotroph pituitary neuroendocrine tumors and silent gonadotroph pituitary neuroendocrine tumors.

[0029] In one embodiment, the silent pituitary neuroendocrine tumor subtype is silent adrenocorticotropic hormone pituitary neuroendocrine tumor and silent gonadotropin pituitary neuroendocrine tumor.

[0030] In another aspect, the present application provides a method for diagnosing a silent pituitary neuroendocrine tumor, comprising the step of detecting the expression level or methylation level of METTL7A in a sample.

[0031] In one embodiment, the method specifically comprises:

[0032] 1) collecting a plasma sample or a tumor tissue sample to be tested;

[0033] 2) amplifying METTL7A in the plasma sample or the tumor tissue sample by PCR with specific primers;

[0034] 3) detecting the amplification effect by agarose gel electrophoresis;

[0035] 4) judging the silent pituitary neuroendocrine tumor subtype of the sample according to the detection result.

[0036] In one embodiment, the step 2) comprises the step of amplifying the methylation site cg05097930 of METTL7A by PCR with specific primers.

[0037] In another aspect, the present application provides a reagent or kit for distinguishing silent adrenocorticotropic hormone pituitary neuroendocrine tumor and silent gonadotropin pituitary neuroendocrine tumor, comprising a reagent for detecting the expression level and / or methylation level of METTL7A.

[0038] In another aspect, the present application provides a reagent or kit for identifying the silent pituitary neuroendocrine tumor subtype, comprising a reagent for detecting the expression level and / or methylation level of METTL7A.

[0039] In another aspect, the present application provides a method for distinguishing silent adrenocorticotropic hormone pituitary neuroendocrine tumor and silent gonadotropin pituitary neuroendocrine tumor, comprising the step of detecting the expression level and / or methylation level of METTL7A in a sample.

[0040] In one embodiment, the method is one or several of methylation-specific PCR (MSP), methylation sequencing, methylation chip or pyrosequencing.

[0041] Preferably, the method is a methylation-specific PCR (MSP) method.

[0042] In one embodiment, the method comprises the following steps:

[0043] 1) collecting a plasma sample or a tumor tissue sample to be tested;

[0044] 2) amplifying METTL7A in the plasma sample or the tumor tissue sample by specific primers through PCR;

[0045] 3) detecting the amplification effect by agarose gel electrophoresis;

[0046] 4) judging the silent pituitary neuroendocrine tumor subtype of the sample according to the detection result.

[0047] In an embodiment, the step 1) is collecting plasma in the sample to be tested; preferably, collecting a plasma ctDNA (circulating tumor DNA) sample in the sample to be tested;

[0048] In an embodiment, the step 2) is amplifying METTL7A in the plasma by specific primers through PCR.

[0049] Preferably, the step 2) comprises a step of performing PCR amplification by using a PCR kit; preferably, the specific primer pair sequence is shown in SEQ ID No. 1-2; preferably, the step 2) comprises a step of amplifying METTL7A methylation site cg05097930 by specific primers through PCR.

[0050] In an embodiment, the method for judging the silent pituitary neuroendocrine tumor subtype of the sample is that, when the METTL7A methylation result is positive (i.e., the METTL7A methylation result is methylation), the sample is a silent gonadotropin pituitary neuroendocrine tumor.

[0051] In an embodiment, the METTL7A methylation result is negative (i.e., the METTL7A methylation result is unmethylation), the sample is a silent adrenocorticotropin pituitary neuroendocrine tumor.

[0052] In an embodiment, the method for judging the invasiveness of the sample is that, when the METTL7A methylation result is positive (i.e., the METTL7A methylation result is methylation), the invasiveness of the sample is weak.

[0053] In an embodiment, the METTL7A methylation result is negative (i.e., the METTL7A methylation result is unmethylation), the invasiveness of the sample is strong.

[0054] In one embodiment, the method for determining the silent pituitary neuroendocrine tumor subtype of a sample is that, when the METTL7A methylation result is positive (i.e., the METTL7A methylation result is methylation), the sample is a weakly invasive silent gonadotropin pituitary neuroendocrine tumor.

[0055] In one embodiment, the METTL7A methylation result is negative (i.e., the METTL7A methylation result is unmethylation), the sample is a strongly invasive silent adrenocorticotropin pituitary neuroendocrine tumor.

[0056] In one embodiment, the METTL7A methylation level of the sample is elevated, the sample is a weakly invasive silent gonadotropin pituitary neuroendocrine tumor.

[0057] In one embodiment, the elevated METTL7A methylation level of the sample refers to the METTL7A methylation level of the sample compared with that of a normal sample and / or a sample with a silent adrenocorticotropin pituitary neuroendocrine tumor.

[0058] In one embodiment, the elevated METTL7A methylation level of the sample refers to that the METTL7A methylation degree of the sample is higher than that of a normal sample and / or a sample with a silent adrenocorticotropin pituitary neuroendocrine tumor, for example, 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times of the METTL7A methylation degree of a normal sample and / or a sample with a silent adrenocorticotropin pituitary neuroendocrine tumor, or more than 10 times of the METTL7A methylation degree of a normal sample and / or a sample without a silent gonadotropin pituitary neuroendocrine tumor.

[0059] A person skilled in the art can determine the normal METTL7A methylation range according to the METTL7A methylation level of a normal sample and / or a sample with a silent adrenocorticotropin pituitary neuroendocrine tumor. When the METTL7A methylation level of the sample to be tested is higher than the normal METTL7A methylation range, or when the METTL7A methylation level of the sample to be tested is 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times of the upper limit of the normal METTL7A methylation range, or even more than 10 times of the upper limit of the normal METTL7A methylation range, the sample to be tested is a sample of a silent gonadotropin pituitary neuroendocrine tumor.

[0060] In another aspect, the present application provides a method for identifying a silent pituitary neuroendocrine tumor subtype, the method comprising the step of detecting the expression level and / or methylation level of METTL7A in a sample.

[0061] Advantageous effects

[0062] The present application provides a method for distinguishing a silent adrenocorticotropic pituitary neuroendocrine tumor and a silent gonadotropin pituitary neuroendocrine tumor by detecting the expression level and methylation level of METTL7A in a sample, which can be used for preoperative diagnosis and evaluation of the invasiveness of the tumor. BRIEF DESCRIPTION OF DRAWINGS

[0063] Figure 1 Omics screening differential molecule Venn diagram.

[0064] Figure 2 Expression of METTL7A at mRNA and protein levels in SCAs group and SGAs group.

[0065] Figure 3 METTL7A methylation level is increased in SGAs tumor tissue.

[0066] Figure 4 METTL7A methylation level is increased in SGAs plasma tissue.

[0067] Figure 5 METTL7A plasma ctDNA methylation ROC curve. DETAILED DESCRIPTION

[0068] The present application will be further described below in conjunction with the examples. The following description is only a preferred embodiment of the present application and is not intended to limit the present application in other forms. Any person skilled in the art can modify the above disclosed technical content into equivalent embodiments. Any simple modification or equivalent change made in accordance with the technical essence of the present application without departing from the scope of the present application falls within the protection scope of the present application.

[0069] Example 1, screening of target

[0070] Eight samples of silent corticotroph adenomas (SCAs) and seven samples of silent gonadotroph adenomas (SGAs) were collected for differential molecular screening. Specifically, the samples of SGAs with relatively poor invasiveness and relatively low clinical tumor recurrence rate were used as a control group, and the samples of SCAs with relatively high invasiveness and relatively high clinical tumor recurrence rate were used as an experimental group. Bioinformatics analysis was used to screen the most obvious differential molecules between SCAs and SGAs. Batch effects were excluded by principal component analysis (PCA), and DESeq2 was used to screen differential methylation genes (FDR<0.05, FC>2). Key methylation markers were determined by combining LDA (linear discriminant analysis) and heat map clustering.

[0071] Figure 1 A Way chart for differential molecular screening was prepared, and the results showed that there were nine main differential molecules, including METTL7A, CNDP2, ST6GALNAC5, CTSZ, CAST, TBC1D24, SSC5D, and SCG2. Among them, methyltransferase-like 7A (METTL7A, GENEBANK ID: 25840) was the most significant differential molecule.

[0072] Example 2, Verification of Consistency of Expression of METTL7A in Plasma and Tumor Tissue

[0073] In addition, 21 samples of silent corticotroph adenomas (SCAs) and 20 samples of silent gonadotroph adenomas (SGAs) were collected. The expression of METTL7A in the plasma and tumor tissue of the above samples was detected by PCR and WB methods, and the results are shown in Figure 2 . Figure 2 A is the expression of METTL7A at the mRNA level in the plasma and tumor tissue of the above samples detected by the PCR method, Figure 2 B is the expression of METTL7A at the protein level in the plasma and tumor tissue of the above samples detected by the WB method, Figure 2 C is Figure 2 the statistical results of B. It can be seen that the expression of METTL7A in the silent corticotroph adenoma (SCAs) group was significantly higher than that in the silent gonadotroph adenoma (SGAs) group at the mRNA and protein levels.

[0074] The methylation chip and methylation-specific PCR (MSP) method are used to detect the DNA methylation of METTL7A in the tumor tissue of the sample.

[0075] The specific method of high-throughput sequencing by using the methylation chip is as follows: the 935K methylation chip is used for whole genome methylation sequencing of the sample, the sequencing is completed by using the Illumina NextSeq550 system, the data is subjected to quality control by FastQC, the adapter is removed by Cutadapt, and then compared with the human reference genome (HG38), and the differential methylation region (DMR) is screened.

[0076] The specific method of methylation-specific PCR (MSP) detection is as follows:

[0077] The PCR reaction system is configured by referring to the PCR kit, 3-5 μL of the PCR product is taken for 3% agarose gel electrophoresis to detect the amplification effect, and data analysis is performed.

[0078] The PCR reaction solution is prepared according to the following components (the preparation of the reaction solution is carried out on ice). Considering the error in pipetting, the volume of the prepared premix should be at least 10% more than the total volume of all reactions.

[0079] Reaction system Volume (μL) 2x qPCR reaction solution 10 Forward primer (10 μM) 1 Reverse primer (10 μM) 1 Nuclease-free water 6 cDNA 2 Total volume 20

[0080]

[0081] The primer sequences for detecting the METTL7A methylation site cg05097930 are as follows:

[0082]

[0083] The DNA methylation of the tumor tissue is shown in Figure 3 Figure 3 A is the statistical average level of methylation of all hyper- and hypo- DMR regions of the sample, and the abscissa CG represents the silent adrenocorticotropic hormone pituitary neuroendocrine tumor (SCAs), and EG represents the silent gonadotropin pituitary neuroendocrine tumor (SGAs), and the results show that compared with SGAs, 8103 hypermethylated DMR regions and 3158 hypomethylated DMR regions are detected in SCAs; Figure 3 B is a diagram of the methylation of METTL7A promoter, the gray shaded part is the methylation difference part, Lib1-4 represents the silent gonadotropin pituitary neuroendocrine tumor (SGAs), and LibA-C represents the silent adrenocorticotropic hormone pituitary neuroendocrine tumor (SCAs); Figure 3 ​C is the MSP method to detect the methylation of tumor tissue of patients, M represents methylation, U represents unmethylation, I-V are silent gonadotropin pituitary neuroendocrine tumors (SGAs), VI-X are representative of silent adrenocortical pituitary neuroendocrine tumors (SCAs), which indicates that in the tumor tissue, METTL7A is more methylated in SGAs patients than in SCAs patients. Figure 3 It is shown that the METTL7A methylation degree in the silent gonadotropin neuroendocrine tumor (SGAs) group is significantly higher than that in the silent adrenocortical pituitary neuroendocrine tumor (SCAs) group.

[0084] Example 3, correlation of METTL7A methylation with clinicopathology

[0085] The methylation of the plasma DNA of the samples obtained in Example 2 was detected by the method of methylation specific PCR (MSP) in Example 2, and the results are shown in Table 2. Figure 4 Figure 4 In Table 2, M represents methylation, U represents unmethylation, and the first row I-X is the SCAs patient group; the second row I-XI is the SGAs patient group. The results show that the METTL7A methylation degree in the plasma tissue of the silent gonadotropin pituitary neuroendocrine tumor (SGAs) group is significantly higher than that in the silent adrenocortical pituitary neuroendocrine tumor (SCAs) group, which is consistent with the trend of tumor tissue.

[0086] The distribution of the positive rate of METTL7A methylation of the samples obtained in Example 2 was statistically analyzed, and the results are shown in Table 1. The results show that the METTL7A methylation positive rate in the plasma of the silent adrenocortical pituitary neuroendocrine tumor (SCAs) is only 9.52%, while the positive rate of the silent gonadotropin pituitary neuroendocrine tumor (SGAs) group is 90% (p<0.05), which indicates that METTL7A methylation has high specificity for SGAs; in the SCAs group, the Ki-67 index of the methylation negative sample is significantly higher than that of the methylation positive sample (p<0.05), that is, in the SCAs group, the degree of METTL7A methylation is related to the invasiveness, and the lower the methylation degree, the higher the invasiveness; in the SGAs group, the methylation positive sample is a low invasiveness subtype (Ki-67<3%), which is consistent with the pathological results at a rate of 95% (Ki-67>3% indicates high invasiveness, and Ki-67<3% indicates low invasiveness).

[0087] Table 1 Positive rate distribution of METTL7A methylation in different pathological categories

[0088]

[0089] ​

[0090] The data were statistically analyzed by using SPSS 24.0 statistical software. Through the analysis of the methylation degree of METTL7A and the pathological category in Table 1, the ROC curve (Receiver operating characteristic curve) was drawn, which is mainly used to evaluate the effect of a certain index on the classification / diagnosis of two types of subjects, and to find the best index critical value to make the classification effect the best. The ROC curve is a curve drawn with FPR (False positive rate) as the abscissa and TPR (True positive rate) as the ordinate. The results are shown in Figure 5 The area under the ROC curve can reach 0.902. Therefore, the methylation degree of METTL7A can be used as an index to distinguish the silent gonadotropin adenoma (SGAs) group and the silent adrenocortical hormone adenoma (SCAs) group.

[0091] The above results show that the METTL7A methylation level of the silent gonadotropin pituitary neuroendocrine tumor (SGAs) group is significantly higher than that of the silent adrenocortical hormone pituitary neuroendocrine tumor (SCAs) group, and is negatively correlated with tumor invasiveness (such as cavernous sinus invasion, high Ki-67 index). The METTL7A methylation modification of the silent gonadotropin pituitary neuroendocrine tumor (SGAs) group affects the expression of METTL7A, so that the expression of METTL7A in the silent gonadotropin pituitary neuroendocrine tumor (SGAs) group is lower than that in the silent adrenocortical hormone pituitary neuroendocrine tumor (SCAs) group.

[0092] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details in accordance with all the teachings disclosed herein, and these changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. Use of a detection agent for the expression level and / or methylation level of METTL7A (methyltransferase like 7A) in a sample in the preparation of a product for the diagnosis of silent pituitary neuroendocrine tumors.

2. Use according to claim 1, characterized in that, The METTL7A methylation level is the methylation level of METTL7A methylation site cg05097930.

3. Use according to claim 1, characterized in that, The diagnosis includes early screening.

4. Use according to claim 1, characterized in that, The detection agent is one or several detection agents for methylation-specific PCR (MSP), methylation sequencing, methylation chip or pyrosequencing.

5. Use according to any one of claims 1 to 4, characterized in that, The sample is a plasma sample and / or a tumor tissue sample.

6. Use of a METTL7A methylation site as a diagnostic marker for silent pituitary neuroendocrine tumors; preferably, the METTL7A methylation site is METTL7A gene cg05097930 methylation site.

7. A product for the diagnosis of silent pituitary neuroendocrine tumors, characterized in that, The product contains reagents for the detection of the expression level and / or methylation level of METTL7A; preferably, the METTL7A methylation level is the methylation level of METTL7A methylation site cg05097930.

8. The product of claim 7, wherein, The product is a kit.

9. A method of diagnosing a silent pituitary neuroendocrine tumor, characterized in that, The method comprises the step of detecting the expression level and / or methylation level of METTL7A in a sample; preferably, the METTL7A methylation level is the methylation level of METTL7A methylation site cg05097930.

10. The method of claim 9, wherein, The method specifically comprises: 1) Collecting a plasma sample or a tumor tissue sample to be tested; 2) PCR amplification of METTL7A in the plasma sample or the tumor tissue sample by specific primers; 3) Agarose gel electrophoresis to detect the amplification effect; 4) Determine the silent pituitary neuroendocrine tumor subtype of the sample according to the detection results.

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