CN111808950B - Thyroid papillary carcinoma miRNA marker and application thereof - Google Patents
Thyroid papillary carcinoma miRNA marker and application thereof Download PDFInfo
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Abstract
The invention provides a miRNA marker related to thyroid papillary carcinoma, which is miRNA-503-5p; meanwhile, the application of the miRNA marker in preparing a tool for predicting thyroid cancer risk or diagnosing thyroid nipple cancer is provided. According to the invention, the miRNA data of the papillary thyroid cancer in the GEO and TCGA databases are analyzed, miRNA related to the papillary thyroid cancer is screened out, and the expression of the miRNA in a clinical sample is verified by fluorescent quantitative PCR, so that a basis is provided for diagnosis and treatment of the papillary thyroid cancer. The test proves that miRNA-503-5p can effectively distinguish papillary thyroid cancer specimens from normal specimens. The invention provides a new diagnosis method for clinically diagnosing papillary thyroid cancer at the molecular level.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a thyroid papillary carcinoma miRNA marker and application thereof.
Background
Papillary thyroid carcinoma (Papillary thyroid cancer, PTC) is the most common histological type of thyroid carcinoma, accounting for about 90% of thyroid carcinomas, and is the malignant tumor of the endocrine system with the fastest growing incidence in recent years. In recent years, with the increase of thyroid diagnosis means such as ultrasonic examination and fine needle biopsy, the incidence of papillary thyroid cancer has been remarkably increased, and the incidence of papillary thyroid cancer has become the fourth cancer worldwide. Notably, the incidence of thyroid papillary carcinoma tumors >4cm in diameter and localized and distant metastatic tumors also increased, resulting in increased PTC mortality. Surgical excision of papillary thyroid carcinoma presents a number of complications, such as recurrent laryngeal nerve injury and hypoparathyroidism, which have a significant impact on the quality of life of the patient. Therefore, diagnosis and intervention of papillary thyroid cancer are performed early, and the method has very important significance for prognosis and quality of life of patients with papillary thyroid cancer.
PTC is a secret illness and patients often have no special symptoms. PCT is screened clinically mainly by methods such as palpation, ultrasound, nuclide imaging, etc., but the diagnostic level of these methods is still very limited. At present, the cytological biopsy by fine needle puncture under ultrasonic guidance is the most effective examination mode for diagnosing thyroid nodules before operation, but a part of thyroid nodules are difficult to diagnose clearly, and the detection accuracy and sensitivity are low. Therefore, research into new diagnostic methods to improve the diagnostic efficiency of PTC has become a major direction of research.
microRNAs (miRNAs) is a small non-coding RNA of 19 to 25 nucleotides in length involved in epigenetic regulation of gene expression, which binds to the 3' UTR region of mRNA, down regulates protein-encoding gene expression, and inhibits protein synthesis at the translational level. It is estimated that about 1/3 of the genes in organisms are regulated by miRNAs. miRNAs are widely involved in pathophysiological processes of disease (especially in malignant tumors), act as "hinges" in gene regulatory networks, control many targets, and have become a promising new potential target for diagnosis and monitoring of malignant tumor biomarkers and cancer treatment.
Detecting the expression level of miRNA may provide a reference for clinical diagnosis of papillary thyroid cancer. Currently, studies have shown that abnormal miRNA expression will lead to abnormal expression of genes associated with the occurrence of cancer, thereby inducing the occurrence of cancer. Therefore, finding new miRNA markers for diagnosis and detection of papillary thyroid cancer has great significance.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, the miRNA marker related to the papillary thyroid cancer can be used for judging the papillary thyroid cancer tissue sample, and an effective means is provided for the clinical diagnosis and risk prognosis of the papillary thyroid cancer.
The invention also provides application of the miRNA marker in preparation of a tool for predicting thyroid cancer risk or diagnosing thyroid nipple cancer.
The invention also provides application of the miRNA marker in a high-throughput sequencing platform.
The invention also provides a chip for predicting the risk of papillary thyroid cancer or diagnosing papillary thyroid cancer.
The invention also provides a kit for predicting the risk of or diagnosing papillary thyroid cancer.
The invention also provides a product for diagnosis and screening of papillary thyroid cancer.
According to an embodiment of the first aspect of the invention, the miRNA marker is miRNA-503-5p.
Further, the miRNA-503-5p is selected from at least one of the following miRNAs: initial miRNA-503-5p, precursor miRNA-503-5p and mature miRNA-503-5p; the initial miRNA-503-5p is sheared and expressed into mature miRNA-503-5p in human cells; the precursor miRNA-503-5p is sheared and expressed into mature miRNA-503-5p in human cells.
The miRNA marker according to the specific embodiment of the invention has at least the following beneficial effects: miRNA-503-5p is used as a miRNA marker related to thyroid papillary carcinoma, the expression level of the miRNA-503-5p in thyroid papillary carcinoma tissues is higher than that in other tissues, and the miRNA-503-5p is characterized in that the expression level in the tissues of thyroid papillary carcinoma patients with poorer prognosis is higher than that in the tissues of thyroid papillary carcinoma patients with better prognosis, so that the miRNA-503-5p can be used for preparing related tools for predicting the risk of thyroid cancer or diagnosing the thyroid papillary carcinoma, and the diagnosis of the thyroid papillary carcinoma patients can be carried out by detecting the amount of miRNA-503-5p in the tissues.
In some specific embodiments of the invention, the miRNA markers further comprise sequences obtained by base modification or addition of bases at the 5' end of miRNA-503-5p.
It will be appreciated that miRNA-503-5p of the invention includes functional equivalents, i.e., variants, of the constitutive nucleic acid molecules that exhibit the same function as the intact miRNA-503-5p nucleic acid molecules, although they are mutated by deletion, substitution or insertion of nucleotide residues.
It is well known in the art that to ensure the stability of miRNA, a protective base, such as TT, may be added at one end of the miRNA, or the miRNA base may be modified, but the function of the miRNA is not affected. Therefore, it is well known to those skilled in the art that the sequence obtained by base modification of miRNA-503-5p or addition of a base at the 5' end without affecting the function of miRNA-503-5p is also included in the scope of the present invention.
Further, the miRNA-503-5p is mature miRNA-503-5p, the nucleotide sequence of the mature miRNA-503-5p is shown as SEQ ID NO.1 in a sequence table, and the sequence of the SEQ ID NO.1 is: UAGCAGCGGGAACAGUUCUGCAG.
Although mature miRNA-503-5p is used in some embodiments, it is contemplated by those skilled in the art that the use of the initial miRNA (pi-miRNA-503-5 p), the precursor miRNA (pre-miRNA-503-5 p), will achieve the same technical effect as mature miRNA-503-5p, as the cells have the ability to further process the initial miRNA (pi-miRNA-503-5 p) or the precursor miRNA (pre-miRNA-503-5 p) into mature miRNA-503-5p. The nucleotide sequence of the precursor miRNA-503-5p is shown as SEQ ID NO.2 in a sequence table, and the sequence of SEQ ID NO.2 is as follows: UGCCCUAGCAGCGGGAACAGUUCUGCAGUGAGCGAUCGGUGCUCUGGGGUAUUGUUUCCGCUGCCAGGGUA.
The miRNA-503-5p nucleic acid molecules of the invention may exist in single-stranded or double-stranded form. Mature miRNA-503-5p is predominantly in single-stranded form, whereas miRNA-503-5p precursors are partially self-complementary to form a double-stranded structure. The nucleic acid molecules of the invention may be in the form of RNA, DNA, PNA, LNA.
According to a second aspect of the invention there is provided the use of miRNA-503-5p in the manufacture of a tool for predicting the risk of thyroid cancer or diagnosing papillary thyroid cancer.
Experiments of the invention prove that: the level of miRNA-503-5p in the tissue with the papillary thyroid carcinoma is significantly higher than that of the tissue without the papillary thyroid carcinoma; thus, if the level of miRNA-503-5p in the papillary thyroid cancer tissue of a subject is significantly increased compared to the level of miRNA-503-5p in the tissue in which papillary thyroid cancer does not occur, the subject can be judged to have developed papillary thyroid cancer, thereby providing a diagnostic basis for the formulation of a clinical treatment regimen.
The invention also provides application of miRNA-503-5p in a tool for judging papillary thyroid cancer. If the level of miRNA-503-5p in the tissue of the subject's papillary thyroid cancer is significantly increased compared to the level of miRNA-503-5p in the tissue in which the papillary thyroid cancer does not occur, it is indicative that the subject is developing papillary thyroid cancer.
Further, the means for predicting the risk of papillary thyroid cancer and determining whether papillary thyroid cancer occurs include, but are not limited to, a kit, a chip, a test paper, etc.
Further, the kit comprises reagents for detecting the expression level of miRNA-503-5p in a test sample. The reagent comprises a primer and/or a probe for detecting miRNA-503-5p.
Further, the reagents comprise reverse transcription primers and/or amplification primers used in qPCR experiments; the reverse transcription primer is an Oligo (dT) specific RT primer; an upstream primer sequence of the amplification primer such as; the downstream primer of the amplification primer is a universal reverse primer.
Further, reagents and enzymes commonly used in PCR technology are also included in the kit.
According to a third aspect of the invention, there is provided the use of the above-described miRNA-503-5p in a high throughput sequencing platform. The expression level of miRNA-503-5p in the papillary thyroid cancer tissue of the sample to be detected can be obtained through high-throughput sequencing, and the result of the sample to be detected is compared with the tissue sample without the papillary thyroid cancer, so that whether the sample to be detected has the risk of the papillary thyroid cancer or whether the sample to be detected has the papillary thyroid cancer can be easily judged. Therefore, the application of obtaining the correlation between miRNA-503-5p and the value papillary carcinoma through high-throughput sequencing is also included in the protection scope of the invention.
The chip according to the fourth aspect of the invention, the chip comprising a solid support; and an oligonucleotide probe immobilized on the solid support, the oligonucleotide probe comprising a portion or all of a sequence that specifically corresponds to miRNA-503-5p. The oligonucleotide probes may also include oligonucleotide probes against mirnas that have been reported in the prior art to be useful in determining whether papillary thyroid cancer has occurred. The condition that the detection probes of the miRNAs are placed on the same chip to jointly judge the papillary thyroid cancer by detecting the miRNA indexes is also included in the protection scope of the invention.
Further, the solid phase carriers include various materials commonly used in the field of gene chips, such as, but not limited to, nylon membranes, glass or silicon wafers modified with active groups (e.g., aldehyde groups, amino groups, etc.), unmodified glass slides, plastic sheets, etc.
In the present invention, the miRNA chip may be prepared by a conventional method for manufacturing a biochip known in the art, for example, when a modified slide or a silicon wafer is used as a solid phase carrier, and an amino-modified poly-dT string is contained at the 5' end of a probe, an oligonucleotide probe may be prepared into a solution, and then spotted on the modified slide or the silicon wafer using a spotter to be arranged into a predetermined sequence or array, and then fixed by standing overnight, thereby obtaining the miRNA chip of the present invention. If the nucleic acid does not contain amino modifications, the preparation method can also be referred to as: wang Shenwu, ind. Infinite Instructions on Gene diagnosis technology-nonradioactive Manual; J.L.erisi, V.R.Iyer, P.O.BROWN.Exploring the metabolic and genetic control of gene expression on a genomic scale.science,1997;278:680 and Ma Liren, jiang Zhonghua Ji, biochip Beijing: chemical industry Press 2000,1-130.
According to a fifth aspect of the invention, the kit comprises reagents for detecting the expression level of miRNA-503-5p in a papillary thyroid cancer tissue of a subject. Comparing the expression level of miRNA-503-5p in the tissue without the occurrence of papillary thyroid cancer, if the expression level of miRNA-503-5p in the papillary thyroid cancer is detected by the kit to be significantly increased, the subject is judged to have a high risk of or has the occurrence of papillary thyroid cancer.
Further, the reagent comprises a primer and/or a probe for detecting the expression level of miRNA-503-5p. The reagents also include other primers and/or probes for mirnas that have been reported in the prior art to be useful in determining the risk of papillary thyroid cancer, or whether papillary thyroid cancer is occurring. The condition that the detection primers and/or probes of the miRNAs are placed in the same kit and the papillary thyroid cancer is jointly judged by detecting the miRNA indexes is also included in the protection scope of the invention.
The miRNA-503-5p of the present invention may be natural or synthetic, or may be obtained by transfecting cells with a vector capable of expressing a DNA fragment of miRNA-503-5p. The vector comprises a viral vector and a eukaryotic vector.
The viral vector may be any suitable vector including, but not limited to, retroviral vectors, adenoviral vectors, adeno-associated viral vectors, herpes viral (e.g., herpes simplex, vaccinia and EB virus) vectors, alphaviral vectors.
Eukaryotic expression vectors may be any suitable expression vector including, but not limited to, pCMV-Myc expression vectors, pcdna3.0 expression vectors, pcdna3.1 expression vectors, pEGFP expression vectors, pEF Bos expression vectors, pTet expression vectors, pTRE expression vectors, or vectors engineered on the basis of well-known expression vectors, such as pBin438, pCAMBIA1301, etc.
The DNA fragment capable of expressing miRNA-503-5p can be obtained as follows: searching the position and specific sequence information of miRNA-503-5p on a genome from an miRNA database (http:// microrner.ac.uk/sequences /), determining the position of miRNA-503-5p initial miRNA according to the genome sequence, designing specific primers in the interval of 500-800bp upstream and downstream of the miRNA-503-5p initial miRNA position, and amplifying the sequence in the middle of the primers to obtain the DNA fragment for expressing miRNA-503-5p.
A product according to the sixth aspect of the invention for detecting the expression level of miRNA-503-5p; the product is any one of the following 1) to 4):
1) Diagnosing or assisting in diagnosing whether the sample to be tested is a papillary thyroid cancer sample;
2) Screening or assisting in screening whether the sample to be tested is a product of a papillary thyroid cancer sample;
3) Diagnosing or assisting in diagnosing whether the person to be tested is a thyroid papillary carcinoma patient;
4) Screening or assisting in screening whether the person to be tested is a product of a papillary thyroid cancer patient.
Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a graph showing the expression level of miRNA-503-5p in papillary thyroid carcinoma tissue and paracancerous tissue in the TCGA database according to example 1 of the present invention;
FIG. 2 is a graph showing the expression level of miRNA-503-5p in the papillary thyroid carcinoma tissue and the paracancerous tissue in the data set GSE113629 of the GEO database of example 1;
FIG. 3 is a schematic diagram showing the correlation of miRNA-503-5p and prognosis with clinical data and expression data in the TCGA database of example 1 of the present invention;
FIG. 4 is a graph showing the real-time fluorescence quantitative PCR detection of the expression level of miRNA-503-5p in papillary thyroid carcinoma tissues and paracancerous tissues in example 2 of the present invention.
Detailed Description
The invention is further illustrated below in conjunction with specific examples, which are provided solely for the purpose of illustrating the invention and are not meant to limit the scope of the invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1: screening of miRNA related to papillary thyroid carcinoma
1. Thyroid papillary carcinoma data retrieval
The method comprises the steps of constructing a search term ("thyroid AND cancer" [ MeSH terminals ] OR "papillary AND thyroid AND cancer" [ All Fields ]) AND ("gse" [ Filter ] AND "[ Organism ]), AND searching in an NCBI GEO (Gene Expression Omnibus) database according to a preset sample screening strategy to obtain 1 set of thyroid papillary carcinoma MiNRA data sets. The TCGA (The Cancer Genome Atlas, cancer genome map) data was obtained by downloading all data of papillary thyroid carcinoma (tumor code THCA) via GDCRNATools package in R language environment. A preset sample screening strategy: the limiting study type was "non-coding RNAprofiling by array", and a dataset meeting the following criteria would be included in our study: (1) the selected dataset must simultaneously include whole genome miRNA expression data; (2) these data are from biopsies of the papillary thyroid case group and the control group; (3) the study considered either the normalized or the original dataset.
2. Thyroid papillary carcinoma miRNA expression data integration analysis
Performing background correction and standardization on 1 set of thyroid cancer miNRAGEO data and TCGA thyroid cancer data through transcriptome data analysis software, then performing t-test to obtain P values, calculating an effector, combining the P values by Fisher test, combining the effector by adopting a random effect model, screening differential expression miRNAs, searching intersection, screening out 29 differential expression miRNAs, wherein 19 genes with up-regulated expression levels and 10 genes with down-regulated expression levels. Based on searching databases such as universal, known network, pubmed, science and the like, the miRNAs verified in the literature are screened out from 19 upregulated miRNAs, 3 upregulated miRNAs are determined to carry out RT-qPCR verification, and finally the miRNA-503-5p with the best expression is selected as a target molecule.
3. Results
FIG. 1 is a graph showing the expression level of miRNA-503-5p in papillary thyroid carcinoma tissues and paracancerous tissues in the TCGA database of this example. As the TCGA data results in fig. 1 show, miRNA-503-5p expression levels in papillary thyroid carcinoma tissues were significantly increased compared to the paracancestral thyroid carcinoma tissues (p < 0.0001). The results of the T test calculation using two samples by GraphPad Prism8 software showed that the P-value < = 0.05, the elevation was considered statistically significant, whereas the P-value < = 0.0001 for miRNA-503-5P, so that the cancer tissue was considered significantly elevated relative to the paracancerous tissue.
FIG. 2 is a graph showing the expression level of miRNA-503-5p in the tissue of papillary thyroid carcinoma and the tissue beside the carcinoma in the data set GSE113629 of the GEO database of the present example; as shown by GEO data results in fig. 2, miRNA-503-5p expression levels in papillary thyroid carcinoma tissues were significantly increased compared to paracancestral thyroid carcinoma tissues (p < 0.0001).
From the results shown in fig. 1 and 2, it can be demonstrated that the miRNA-503-5p obtained by screening can be applied to the detection of papillary thyroid cancer tissues, and since the expression of miRNA-503-5p has significant differences in the papillary thyroid cancer tissues and the paracancerous tissues, whether the detected tissues are papillary thyroid cancer tissues or not can be judged by judging the expression level of miRNA-503-5p, thereby diagnosing whether the papillary thyroid cancer is caused.
4. Correlation analysis of miRNA-503-5p and prognosis
Using TCGA expression data and clinical information, the association between miRNA-503-5p expression and patient OS (Overall Survival) was assessed. Patients were cut off at the median survival time of each sample, and the patients were divided into high-expression and low-expression groups, and survival differences were evaluated using the survivintype package in the R language.
FIG. 3 is a schematic diagram showing the correlation of miRNA-503-5p with prognosis in clinical data and expression data in TCGA databases according to the embodiments of the present invention; as shown in fig. 3, miRNA-503-5p is highly expressed, and patient survival rate is low; miNRA-503-5p is expressed in a low mode, and the survival rate of patients is high. miRNA-503-5p was inversely related to patient prognosis with statistical difference (×p=0.031). Thus, as can be seen from the results of FIG. 3, the prognosis of a patient with papillary thyroid cancer can be predicted by detecting the expression level of miRNA-503-5p in the tissue of the patient.
Example 2: real-time fluorescence quantitative PCR (polymerase chain reaction) verification of differentially expressed miRNA-503-5p
1. miRNA extraction
An equal volume of lysate was added to every 200 μl of serum using the Tiangen miRNA extraction kit, and mixed for 30 seconds with shaking by a shaker. After 5min at room temperature, centrifugation at 12,000rpm for 10min, the supernatant was taken, 200 μl chloroform was added, shaking vigorously for 15 seconds, after 5min at room temperature, centrifugation at 12,000rpm for 15min, the sample was divided into three layers: the yellow organic phase, the middle layer and the colorless aqueous phase, RNA is mainly in the aqueous phase, the aqueous phase is transferred to a new tube, absolute ethyl alcohol with the volume of 1/3 of the transfer solution is slowly added and is uniformly mixed, the mixture is transferred to an adsorption column, and the mixture is placed at room temperature for 2min and centrifuged at 12,000rpm for 30 seconds, so that effluent liquid is reserved. Slowly adding 2/3 of the volume of absolute ethyl alcohol, mixing, transferring into an adsorption column, standing at room temperature for 2min, centrifuging at 12,000rpm for 30 seconds, and retaining the adsorption column after centrifuging. 500. Mu.l of deproteinized solution was added to the adsorption column, and the mixture was centrifuged at 12,000rpm at room temperature for 30sec, and the waste solution was discarded. 500 μl of the rinse solution was centrifuged at 12,000rpm for 30 seconds at room temperature. The column was placed in a 2ml collection tube and centrifuged at 12,000rpm for 1min at room temperature to remove residual liquid. The column was then transferred to a new 1.5ml centrifuge tube, 15-30. Mu.l RNase-free water was added and centrifuged at 12,000rpm for 2min at room temperature.
2. Reverse transcription
10 pg-1. Mu.g of RNA template was mixed with 2. Mu.l of 10-fold buffer, 2. Mu.l of dATP (10 mM), 0.5. Mu.l of primer (nucleotide sequence shown as SEQ ID NO.3 in the sequence Listing; sequence TAGCAGCGGGAACAGTTCTGCAG; sequence), 0.5. Mu.l of ribonuclease inhibitor and water, and finally incubated at 37℃for 1h in a volume of 20. Mu.l. Then 1. Mu.l of 0.5. Mu.g/. Mu.l of specific RT primer was added to the reaction tube and incubated on ice for at least 2min immediately after incubation at 70℃to disrupt the secondary structure of RNA and primer. Finally, the 20. Mu.l reaction mixture was mixed with 4. Mu.l of 5-fold buffer, 1. Mu.l of dNTP (10 mM), 0.5. Mu. l M-MLV reverse transcriptase, 0.5. Mu.l of ribonuclease inhibitor, 10. Mu.l of polyA reaction mixture and 4. Mu.l of ribonuclease-free water, and incubated at 42℃for 1 hour.
3. Real-time fluorescent quantitative PCR detection
With a 25. Mu.l reaction system, 3 parallel tubes were set for each sample, and all amplification reactions were repeated more than three times to ensure reliability of the results. The following reaction system was prepared: SYBR Green polymerase chain reaction system 12.5. Mu.l, forward primer (5. Mu.M/l) 1. Mu.l, reverse primer (5. Mu.M/l) 1. Mu.l (the invention uses tailing primer design, the nucleotide sequence of the forward primer is shown as SEQ ID NO.3 in the sequence table, the sequence is TAGCAGCGGGAACAGTTCTGCAG, the reverse primer is universal sequence primer), template cDNA 2. Mu.l, and no enzyme water 8.5. Mu.l. Each operation was performed on ice. The amplification procedure was: 10min at 95℃and 40 cycles (20 s at 95℃and 55s at 60 ℃). The PCR reaction was performed on a fluorescent real-time quantitative PCR instrument using SYBR Green as a fluorescent label. The target band is determined through melting curve analysis and electrophoresis, and the relative quantification is carried out by a delta CT method.
4. Results
As shown in table 1, it can be seen that the expression level of miRNA-305-5p in 11 papillary thyroid cancer tissues was up-regulated compared to the paracancerous tissues, and there was a significant difference compared to the positive control miRNA-31-5p, miRNA-3065-5 p). The miRNA-31-5p and the miRNA-3065-5p are known papillary thyroid cancer markers, and compared with the known papillary thyroid cancer markers, the marker miRNA-305-5p provided by the invention has more obvious difference, so that the marker miRNA-305-5p provided by the invention has higher accuracy and sensitivity when used for detecting the papillary thyroid cancer.
Table 1 shows the expression levels of the target genes (relative to RNU 6B) in the respective samples
FIG. 4 is a graph showing the real-time fluorescence quantitative PCR detection of the expression level of miRNA-503-5p in papillary thyroid carcinoma tissue and paracancestral thyroid carcinoma tissue, and the ordinate represents the relative expression level of miRNA-503-5p. As shown in fig. 4, the miRNA-503-5p content in papillary thyroid cancer tissue was significantly higher than that in paracancerous tissue (< 0.001 p), thus proving that miRNA-503-5p can be used as a marker for detecting papillary thyroid cancer. Statistical treatment was performed with Gram Pad Prism8 software the data of table 1.
In summary, the invention analyzes the miRNA data of papillary thyroid cancer in GEO and TCGA databases, screens miRNA related to papillary thyroid cancer, verifies the expression of the miRNA in clinical samples by using fluorescent quantitative PCR (RT-qPCR), and provides basis for diagnosis and treatment of papillary thyroid cancer. Experiments prove that the miRNA-503-5p has higher expression level in the papillary thyroid cancer tissues and low expression level in normal tissues, so that the correlation with the papillary thyroid cancer can be judged by measuring the miRNA-503-5p expression level in a sample.
The foregoing description is only illustrative of the present invention and is not intended to limit the scope of the invention, and all equivalent changes made by the specification and drawings of the present invention, or direct or indirect application in the relevant art, are included in the scope of the present invention.
Sequence listing
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Claims (4)
1. An application of miRNA markers related to thyroid papillary carcinoma in preparing products for predicting thyroid cancer risk or diagnosing thyroid papillary carcinoma is provided, wherein the miRNA markers are miRNA-503-5p, and the nucleotide sequence of the miRNA-503-5p is shown as SEQ ID NO. 1.
2. The use according to claim 1, wherein the product comprises a kit, a chip or a test paper.
3. A kit for predicting the risk of or diagnosing papillary thyroid cancer, which is characterized by comprising a reagent for detecting the expression level of miRNA-503-5p in a sample, wherein the nucleotide sequence of the miRNA-503-5p is shown as SEQ ID NO. 1.
4. The kit of claim 3, further comprising primers and/or probes for detecting the expression level of miRNA-503-5p.
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