Application of SCTAG in early diagnosis, prognosis evaluation and recurrence early warning of esophageal cancer

Abstract

The invention belongs to the field of biological medicine, and particularly relates to novel application of SCTAG in diagnosis or prediction (including early diagnosis, prognosis evaluation and recurrence early warning) of esophageal cancer. The research finds that high-level SCTAG exists in tumor tissues and serum of the early esophageal cancer, the five-year lifetime of a patient with the high serum SCTAG level before an operation is poor, and the serum SCTAG after the operation is unchanged or increased compared with the serum SCTAG before the operation indicates the high recurrence risk in a short period, so that the SCTAG can be used as a marker for early diagnosis, prognosis evaluation and recurrence early warning of the esophageal cancer.

Description

Technical Field

[0001] The present application belongs to the field of biomedicine and relates to a new use of SCTAG, and specifically to the application of SCTAG in the diagnosis or prediction of esophageal cancer (especially including disease warning, early diagnosis, prognosis assessment and recurrence warning). Background Art

[0002] Esophageal cancer is a highly prevalent digestive tract malignancy worldwide and one of the most common malignancies in China, which seriously affects people's lives and health. China has the highest incidence of esophageal cancer in the world. According to the pathological type classification, esophageal cancer is mainly divided into two types: esophageal squamous cell carcinoma and esophageal adenocarcinoma. At present, squamous cell carcinoma is the main pathological type of esophageal cancer worldwide. In economically underdeveloped areas, its main pathological type is squamous cell carcinoma, while in many countries in North America and Europe (such as the United States, France, Australia, the United Kingdom, the Netherlands, etc.), its main pathological type is adenocarcinoma.

[0003] The incidence of esophageal cancer in China has its own unique geographical distribution characteristics. The most typical feature is the high incidence rate in the border area of ​​Henan, Hebei and Shanxi provinces in the southern section of Taihang Mountain (the highest incidence rate is in Lin County, Henan). From the perspective of China's three major regions of east, central and west, there are also large differences in the incidence of esophageal cancer. In 2015, the incidence rate in central China was 19.6 / 100,000, higher than that in the eastern region (17.2 / 100,000) and the western region (16.8 / 100,000). Moreover, there are urban-rural differences in the incidence of esophageal cancer, and the incidence rate of esophageal cancer in rural areas is significantly higher than that in urban areas. In terms of gender, the incidence rate of esophageal cancer in men is much higher than that in women.

[0004] The exact cause of esophageal cancer is not yet completely clear. Long-term and repeated physical or chemical damage can lead to an increased incidence of esophageal cancer. At present, smoking and heavy drinking have been proven to be important causes. Other causes may include bad eating habits, such as eating less vegetables and fruits, overeating processed meat and fat, hard and overheated food, eating quickly, etc.; lack of certain trace elements and vitamins, and genetic susceptibility factors for esophageal cancer. There are currently no specific serum markers for esophageal cancer. Common standard tumor markers, such as carcinoembryonic antigen, cancer antigen CA19-9 and CA125, have low diagnostic sensitivity and specificity in esophageal cancer, and are of little value in screening, detecting recurrence, predicting treatment response or survival possibility. However, when patients have obvious symptoms of dysphagia, it often means that esophageal cancer has progressed to the middle and late stages and missed the best treatment time. Therefore, finding a suitable screening program and a more effective treatment method is crucial to improving the prognosis of esophageal cancer patients and prolonging their survival period.

[0005] SCTAG (super-sensitive Cancer / Testis Antigen for Gut diagnosis) is a new type of cancer-testis antigen, which is synthesized in late spermatids of many animals and binds to DNA, causing the sperm cell genome to condense into a genetically inactive state. Under physiological conditions, SCTAG is only expressed in testicular tissue and not in other normal tissues. The application of SCTAG in the diagnosis of esophageal cancer has not been mentioned in the prior art. In view of this, the present application is proposed. SUMMARY OF THE INVENTION

[0007] This application accurately finds evidence of SCTAG expression in early esophageal cancer lesions at the protein level: there is a high level of SCTAG protein expression in the serum of patients with early esophageal cancer, and the increased or unchanged level of SCTAG protein in the serum after surgery indicates a high risk of recurrence after surgery. The above results show that SCTAG can not only be used for the early diagnosis of esophageal cancer, but also has an early warning effect on the recurrence of esophageal cancer prognosis. In addition, this application also detected SCTAG protein in cancer tissues, paracancerous tissues and normal tissues of esophageal cancer patients through immunohistochemistry and found that SCTAG was less positively expressed in non-tumor areas (paracancerous tissues and normal tissues) of esophageal cancer patients, and the closer the tissue is to the central area of ​​esophageal cancer, the higher the expression level of SCTAG protein.

[0008] Therefore, the core purpose of this application is to explore a diagnostic or predictive method for esophageal cancer (especially early esophageal cancer), corresponding products and their applications.

[0009] Specifically, this application proposes the following solutions:

[0010] The present application first provides the use of SCTAG protein or gene as a marker in the diagnosis or prediction of esophageal cancer.

[0011] The present application also provides the use of SCTAG protein or gene as a marker in the preparation of esophageal cancer diagnosis or prediction products.

[0012] The present application also provides the use of a detection agent or component for obtaining the SCTAG level in a sample in the diagnosis or prediction of esophageal cancer.

[0013] The present application also provides the use of a detection agent or component for obtaining the SCTAG level in a sample in the preparation of a product for diagnosing or predicting esophageal cancer.

[0014] The present application also provides a product for diagnosing or predicting esophageal cancer, which comprises a detection agent or component for obtaining the SCTAG level in a sample.

[0015] Furthermore, the diagnosis or prediction includes onset prediction / warning, early screening or diagnosis, auxiliary diagnosis, diagnosis (determining whether the disease exists), prediction or judgment of the severity of the disease, prognosis assessment and recurrence monitoring.

[0016] In some preferred aspects, the diagnosis or prediction particularly includes onset warning, early diagnosis or screening, prognosis assessment or recurrence warning of esophageal cancer.

[0017] Furthermore, the sample source includes one or more of tissue, serum, plasma, whole blood, urine, saliva, pleural effusion, ascites or cell culture supernatant; in some preferred aspects, the sample source particularly includes tissue, serum, plasma or whole blood.

[0018] Furthermore, the above-mentioned level includes the gene (nucleic acid) level or protein level, namely the SCTAG gene level or protein level.

[0019] In some aspects, when it is at the nucleic acid level (including DNA level and RNA level, preferably mRNA level), the nucleic acid level can be obtained by sequencing technology, nucleic acid amplification technology, nucleic acid hybridization technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology;

[0020] Further preferably, the nucleic acid level acquisition method includes but is not limited to any of the following methods: gene sequencing, polymerase chain reaction, isothermal amplification reaction, gene chip, probe hybridization, gel electrophoresis, Northern blotting, nucleic acid mass spectrometry or liquid chromatography. Therefore, in one or more specific embodiments, the detection reagent can include primers that can specifically amplify the SCTAG gene or probes that specifically recognize the SCTAG gene.

[0021] In other aspects, when it is a protein level, the protein level is obtained by sequencing technology, immunological technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology;

[0022] Further preferably, the protein level acquisition method includes but is not limited to any of the following methods: amino acid sequencing, enzyme-linked immunosorbent assay, chemiluminescence, immunochromatography, radioimmunoassay, immunohistochemistry, immunoblotting, flow cytometry, gel electrophoresis, protein spectrum or liquid chromatography. Therefore, in one or more specific embodiments, the protein level is obtained by immunological techniques, wherein the reagent can be an antibody that can specifically bind to the SCTAG protein; more preferably, the anti-antibody is a monoclonal antibody to SCTAG, etc.

[0023] Furthermore, the aforementioned product forms include kit forms, device forms (including chip forms, etc.).

[0024] In some aspects, the product is in the form of a kit, in which case the kit may include, in addition to, for example, reagents for detecting SCTAG protein expression or reagents for detecting SCTAG gene expression, sample processing reagents; further preferably, the sample processing reagents may include: at least one or more of sample lysis reagents, sample purification reagents, and sample nucleic acid or protein extraction reagents.

[0025] Furthermore, the aforementioned reagents can be used for quantitative detection or qualitative detection.

[0026] In some aspects, the detection described in the present application can be based on the detection of SCTAG levels alone to achieve the purpose of diagnosis or prediction of esophageal cancer; in other aspects, the detection described in the present application can also include detecting the levels of other markers related to the diagnosis of esophageal cancer, thereby achieving joint monitoring, which are all within the scope of this application.

[0027] The present application also provides a method for predicting or diagnosing esophageal cancer, comprising the following steps:

[0028] (i) obtaining the level of SCTAG in the subject's sample;

[0029] (ii) comparing the SCTAG level with that of a control sample; wherein a significant difference in the SCTAG level between the subject sample and the control sample is an indication that the subject has or will have esophageal cancer in the future; or,

[0030] (ii) comparing with a set threshold absolute amount; wherein the level of the subject's sample being higher than the threshold absolute amount is an indication that the subject has or will have esophageal cancer in the future.

[0031] In some aspects, when the prediction or diagnosis is an early warning of the disease or an early diagnosis, the subject sample is a sample to be tested, and the control sample is a healthy sample; it can be understood that in some specific ways, when the SCTAG detection value is higher, it indicates that the subject has a higher chance of developing esophageal cancer; when the SCTAG detection value is low or absent, it indicates that the subject has a lower chance of developing esophageal cancer.

[0032] In some aspects, when the prediction or diagnosis is a confirmed diagnosis of a disease, the subject sample is a sample from a patient to be examined, and the control sample is a healthy sample.

[0033] In some aspects, when the prediction or diagnosis is prognosis assessment or recurrence monitoring after treatment, the subject sample is a sample from an esophageal cancer patient after treatment, and the control sample is a sample from an esophageal cancer patient before treatment.

[0034] In some specific aspects, when the recurrence monitoring is postoperative recurrence monitoring, the subject sample is a sample of an esophageal cancer patient who has received postoperative treatment, and the control sample is a sample of an esophageal cancer patient who has received preoperative treatment; the specific method may include the following steps:

[0035] (i) obtaining the level of SCTAG in the postoperative samples of the subjects;

[0036] (ii) comparing the SCTAG level with that of the subject's preoperative control sample; when the SCTAG level in the postoperative sample increases or remains unchanged, it indicates a high risk of esophageal cancer recurrence.

[0037] In one or more embodiments of the present application, the postoperative recurrence monitoring includes: taking patient serum, detecting and comparing the changes in the SCTAG gene content or protein expression in the serum samples before and after surgery, monitoring the dynamic changes of the SCTAG gene content or protein expression, and when the serum SCTAG gene content or protein expression increases or remains unchanged after surgery, it indicates a high risk of recurrence; preferably, further screening can be performed, such as pathological detection of tumor tissue or draining lymph nodes and imaging examinations.

[0038] Further, the subject sample includes one or more of tissue, serum, plasma, whole blood, urine, saliva, pleural effusion, ascites or cell culture supernatant. In some preferred aspects, the sample includes tissue, serum, plasma or whole blood; in some more preferred aspects, the sample is a serum sample.

[0039] Furthermore, the SCTAG level includes the gene (nucleic acid) level or protein level of SCTAG.

[0040] In some aspects, when it is the SCTAG nucleic acid level (such as RNA level, especially mRNA level), the nucleic acid level can be obtained by sequencing technology, nucleic acid amplification technology, nucleic acid hybridization technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology; further preferably, the nucleic acid level acquisition method includes but is not limited to any of the following methods: gene sequencing method, polymerase chain reaction method, isothermal amplification reaction method, gene chip method, probe hybridization method, gel electrophoresis method, Northern blotting method, nucleic acid mass spectrometry or liquid chromatography.

[0041] In other aspects, when it is the SCTAG protein level, the method is to detect the protein level of SCTAG, such as protein abundance, protein expression and even protein activity. Preferably, the protein level is obtained by sequencing technology, immunoassay, electrophoresis technology, biomass spectrometry technology or chromatography technology; further preferably, the protein level acquisition method includes but is not limited to any of the following methods: amino acid sequencing, enzyme-linked immunosorbent assay, chemiluminescence, immunochromatography, radioimmunoassay, immunohistochemistry, immunoblotting, flow cytometry, gel electrophoresis, protein spectrum or liquid chromatography. In some specific embodiments, the protein level is obtained by protein immunoassay, so the detection can be carried out by antigen-antibody binding reaction using antibodies to the SCTAG protein; preferably, the antibody used is a monoclonal antibody to the SCTAG protein, etc.

[0042] The present application also provides a method for detecting a marker in a subject having or suspected of having esophageal cancer in vivo or in vitro, the method comprising the step of determining or detecting the level of SCTAG in a sample from the subject.

[0043] The present application also provides a method for evaluating or screening diagnostic or predictive markers for esophageal cancer, comprising the step of performing correlation or consistency analysis between potential biomarkers and SCTAG.

[0044] The present application also provides a method for in vivo or in vitro screening of esophageal cancer therapeutic agents, the method comprising the step of evaluating the SCTAG level of a sample treated with the therapeutic agent.

[0045] Beneficial effects of this application:

[0046] This application discovered and confirmed for the first time that SCTAG protein can be used for the early diagnosis of esophageal cancer and the monitoring of postoperative recurrence, and has significant clinical value. Specifically: 1) This application confirms that SCTAG can be used as a diagnostic marker for esophageal cancer; in particular, SCTAG is used for early screening or early detection of esophageal cancer (such as early esophageal cancer), with good sensitivity and / or specificity; when the SCTAG detection value is high, it indicates that the probability of esophageal cancer in the detected subject is higher; when the SCTAG detection value is low, it indicates that the probability of esophageal cancer in the detected subject is lower. 2) This application confirms that postoperative SCTAG changes can be used for early warning of esophageal cancer recurrence: when the postoperative SCTAG detection value is higher than that before surgery, it indicates that the probability of esophageal cancer recurrence in the detected subject is higher; when the SCTAG detection value is lower than that before surgery, it indicates that the probability of esophageal cancer recurrence in the detected subject is lower. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0048] Figure 1 , gene detection results of tumor tissues and normal tissues; among them, (A) is the SCTAG mRNA expression result in tumor tissues; (B) is the mRNA expression result in normal tissues.

[0049] Figure 2 , the up-regulated expression of SCTAG mRNA in esophageal cancer tumor tissues and adjacent tissues; (A) is the quantitative analysis result of SCTAG mRNA in multiple esophageal cancer tissues; (B) is the quantitative analysis result of SCTAG mRNA in multiple esophageal adjacent tissues.

[0050] Figure 3 , Immunohistochemistry detected the expression level of SCTAG in normal esophageal tissue (left), paracancerous tissue (middle) and cancer tissue (right).

[0051] Figure 4 , Western-Blot was used to detect the expression level of SCTAG in normal esophageal tissue (N) and cancer tissue (T).

[0052] Figure 5 , the diagnostic results of serum SCTAG levels in patients with esophageal cancer; among them, (A) comparison of serum SCTAG levels between the healthy control group and patients with esophageal cancer at different stages; (B) the diagnostic value of SCTAG in early esophageal cancer; (C) the diagnostic value of SCTAG compared with common tumor markers; p<0.05, with statistically significant difference.

[0053] Figure 6 , the results of serum SCTAG level in predicting high recurrence risk after esophageal cancer surgery; among them, (A) the dynamic changes of serum SCTAG before and after surgery; (B) the correlation between preoperative and postoperative serum SCTAG changes and esophageal cancer recurrence; (C) the correlation between preoperative SCTAG level and the prognosis of esophageal cancer patients; p<0.05, with statistical significance.

[0054] Figure 7 , CT images of esophageal cancer recurrence in the groups with decreased (left) and increased (right) serum SCTAG before and after surgery. DETAILED DESCRIPTION OF THE INVENTION

[0056] The technical solution of the present application will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present application.

[0057] The following terms or definitions are provided only to help understand the present application. These definitions should not be construed as having a scope less than that understood by those skilled in the art.

[0058] Unless otherwise defined below, the meaning of all technical terms and scientific terms used in the specific embodiments of the present application is intended to be the same as those generally understood by those skilled in the art. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are still set forth to better explain the present application.

[0059] As used in this application, the terms "include", "comprising", "having", "containing" or "involving" are inclusive or open-ended and do not exclude other unrecited elements or method steps. If a group is defined below as comprising at least a certain number of embodiments, this should also be understood to disclose a group that preferably consists only of these embodiments.

[0060] When referring to a singular noun an indefinite or definite article e.g. "a" or "an", "the" is used, this includes a plural of that noun.

[0061] The terms "approximately" and "substantially" in this application represent the accuracy range that can be understood by those skilled in the art to still ensure the technical effect of the feature in question. The term usually represents ±10% deviation from the indicated value, preferably ±5%.

[0062] Details of this application

[0063] 1. Diagnostic or predictive use

[0064] This application is based on the fact that SCTAG is an oncogene that is abnormally expressed during the formation of esophageal cancer and has important biological functions. This application has carried out detection of SCTAG protein in paracancerous tissues and normal tissues of patients with esophageal cancer, and found that the positive expression of SCTAG protein in non-tumor areas (paracancerous tissues and normal tissues) of patients with esophageal cancer is low, and the expression frequency of SCTAG mRNA in esophageal cancer tumor tissues is higher; in addition, this application has conducted an enzyme-linked immunosorbent assay on the serum of patients with esophageal cancer and healthy subjects, and found that SCTAG expression is also significantly increased in early esophageal cancer. These results show that SCTAG can be used for the early diagnosis of esophageal cancer, that is, it has an early warning prompt effect on the occurrence of early esophageal cancer. This application also compares the levels of serum SCTAG before and after surgery, and finds that the risk of postoperative recurrence in patients with esophageal cancer whose serum SCTAG is increased after surgery is significantly higher than that in patients with esophageal cancer whose serum SCTAG is decreased after surgery.

[0065] Based on these experimental data, the present application can conclude that SCTAG can be used as a diagnostic or predictive marker for esophageal cancer, so the present application at least includes the following applications.

[0066] 1. Application of SCTAG as a marker in the diagnosis or prediction of esophageal cancer.

[0067] 2. Application of SCTAG as a marker in the preparation of diagnostic or predictive products for esophageal cancer.

[0068] 3. Application of SCTAG level detection agents or components in the diagnosis or prediction of esophageal cancer.

[0069] 4. Application of SCTAG level detection agents or components in the preparation of esophageal cancer diagnosis or prediction products.

[0070] The "esophageal cancer" mentioned in this article is also called esophageal cancer, which refers to a malignant tumor originating from the esophageal epithelium. "Early esophageal cancer" usually refers to the invasion of cancer cells limited to the mucosal layer and the mucosal lamina propria, as well as above the submucosal layer, without invading the basal layer, collectively referred to as early esophageal cancer. Gastroenterologists divide the esophagus into 5 layers, and the first and second layers, or the upper 1 / 3 of the third layer, are considered to be early esophageal cancer. At this level, minimally invasive resection under digestive endoscopy can control the risk of lymph node metastasis to less than 25%, especially for the first layer of esophageal cancer, the risk of metastasis can be controlled below 9%. The "diagnosis of early esophageal cancer" mentioned in this article has the same meaning as "early diagnosis of esophageal cancer."

[0071] The "SCTAG" protein described in this article is also known as protamine 1 (PRM1). "SCTAG" and "PRM1" are equivalent in this article. It is a newly discovered cancer-testis antigen (CTA). The SCTAG gene is located on human chromosome 16, with a total length of 399 bp, 1 exon and 2 introns, encoding 51 amino acids, rich in arginine, and positively charged. Protamine is the main protein that binds to DNA in the sperm nucleus. During the elongated spermatid stage of spermatogenesis, human sperm chromatin undergoes a complex transition in which histones are extensively replaced by protamines. This is a carefully regulated transition that includes histone modifications and intermediate and transient replacement of histones by sperm-specific transition proteins. Replacing most histones with SCTAG can promote higher-order chromatin packaging, which is required for normal sperm function. Most patients with abnormal SCTAG levels have increased protamine transcript levels in mature spermatids, suggesting that there may be transcriptional or translational defects. The regulation of SCTAG expression is unique and includes multiple possible mechanisms that may be responsible for dysregulated SCTAG expression and a broad spectrum of defects during spermatogenesis. During spermatogenesis, successful replacement of histone-PRMs is important for normal sperm development, and abnormal SCTAG expression or abnormal SCTAG / 2 ratio can cause male infertility.

[0072] As used herein, the term "diagnosis" generally refers to the process of identifying a medical disorder or disease (such as esophageal cancer) through its signs, symptoms, and especially the results of various diagnostic processes (including detecting the level of SCTAG protein in a biological sample (such as serum) obtained from an individual). As used herein, the term "diagnosis" may include prediction of the future onset of the disease, early screening or diagnosis, auxiliary diagnosis, diagnosis (determining whether the disease exists), prediction or judgment of the severity of the disease, prognosis assessment, monitoring treatment, such as monitoring disease deterioration or recurrence during treatment, evaluating the therapeutic efficacy of the disease, and the selection of a given treatment regimen. Without limitation, in some specific embodiments of the present application, the present application has fully confirmed that the SCTAG level (especially the serum protein expression level) can be used to determine whether the subject has esophageal cancer, and can then be used for early screening, auxiliary diagnosis and diagnosis; in other specific embodiments, the present application also confirms that the unchanged or increased serum SCTAG after surgery indicates a high risk of recurrence in the short term, so the SCTAG level can be used to predict and judge the possibility of recurrence of esophageal cancer after surgery, and can then be used for prognosis assessment and recurrence detection.

[0073] In the uses described in this application, there is no limitation on sample type, sample status, sample processing, or detection method.

[0074] For example, in some embodiments, the sample source can be one or more of tissue, serum, plasma, whole blood, urine, saliva, pleural effusion, ascites or cell culture supernatant selected from the test subject. According to some specific embodiments of the present application, the sample source includes tissue, serum, plasma or whole blood. According to some specific embodiments of the present application, the sample is especially a serum sample. According to some more specific embodiments of the present application, when in postoperative recurrence assessment, the sample is a preoperative and postoperative serum sample of an esophageal cancer patient.

[0075] The sample state of the present application can be in vivo or in vitro; no matter what kind of pretreatment the sample undergoes, as long as the ultimate goal is to detect the SCTAG level of the sample, it is within the scope of the present application.

[0076] The detection agent or component for obtaining the SCTAG level in the sample can be understood to include a detection reagent for directly obtaining the SCTAG level in the sample, or a component for indirectly obtaining the SCTAG level in the sample (such as a chip, setting a computer program to directly obtain an indicator of the SCTAG level in a sample that has been detected).

[0077] The level of SCTAG in the sample can be obtained by obtaining a marker level indicator at the nucleic acid level or protein level. There is no limitation on the acquisition method or detection method, and any method that can be used to directly or indirectly evaluate the level of SCTAG nucleic acid (especially RNA) or protein is suitable for this application.

[0078] It is understood that nucleic acid levels can include DNA levels or RNA levels. There are many ways to detect DNA or RNA levels in this area, including but not limited to sequencing technology, nucleic acid amplification technology, nucleic acid hybridization technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology, all of which can be used in this application. In some ways of this application, include but are not limited to any of the following specific methods: gene sequencing, polymerase chain reaction, isothermal amplification reaction, gene chip method, probe hybridization method, gel electrophoresis, Northern blotting, nucleic acid mass spectrometry or liquid chromatography.

[0079] For example, in some embodiments, the nucleic acid sequencing method described in the present application can be transcriptome sequencing or genome sequencing. In other embodiments of the present application, the nucleic acid sequencing method is high-throughput sequencing, also known as second-generation sequencing ("NGS"). NGS is different from "Sanger sequencing" (first-generation sequencing), which is the electrophoretic separation of chain termination products based on a single sequencing reaction. NGS is a revolutionary change to traditional Sanger sequencing technology, and can sequence hundreds of thousands to millions of nucleic acid molecules at a time. The sequencing platform of the available NGS of the present application is commercially available, including but not limited to Roche / 454FLX, Illumina / Solexa Genome Analyzer and Applied Biosystems SOLID system, etc. Transcriptome sequencing can also quickly and comprehensively obtain nearly all transcripts and gene sequences of a certain species specific cell or tissue in a certain state through a second-generation sequencing platform, which can be used to study gene expression, gene function, structure, alternative splicing and new transcript prediction, etc. In some other embodiments of the present application, the nucleic acid sequencing method can be single-molecule real-time sequencing. Single-molecule DNA sequencing technology is a new generation of sequencing technology developed in the past 10 years, also known as the third-generation sequencing technology, including single-molecule real-time sequencing, true single-molecule sequencing, single-molecule nanopore sequencing and other technologies.

[0080] For example, in other embodiments, the polymerase chain reaction method of the present application includes primers for SCTAG gene amplification, wherein the primer refers to a primer that can specifically amplify the SCTAG encoding gene or mRNA, such as a polynucleotide of a certain length, such as a primer of about 35 nucleotides or longer, which can hybridize with at least a portion of the template sequence and serve as a starting site for synthesizing primer extension products. In some specific embodiments, the probe hybridization method includes a probe of SCTAG, wherein the probe refers to a probe that can specifically identify the SCTAG encoding gene, which is a molecule that can bind to a specific sequence or subsequence or other part of another molecule, generally referring to a nucleic acid probe that binds to another nucleic acid (i.e., a specific sequence of SCTAG as a target nucleotide) through complementary base pairing.

[0081] The detection agent performs detection at the protein level. It can be understood that there are many ways to detect protein in the art, including but not limited to sequencing technology, immunoassay technology, electrophoresis technology, biomass spectrometry technology or chromatography technology, all of which can be used in the present application.

[0082] In some specific embodiments of the present application, it includes but is not limited to any of the following specific methods: amino acid sequencing, enzyme-linked immunosorbent assay, chemiluminescence, immunochromatography, radioimmunoassay, immunohistochemistry, immunoblotting, flow cytometry, gel electrophoresis, protein spectrum or liquid chromatography.

[0083] In some preferred embodiments, the detection agent is used to perform any of the following methods: biological mass spectrometry, amino acid sequencing, electrophoresis, and detection with specific antibodies. The method of detection with specific antibodies can further be immunoprecipitation, immunocoprecipitation, immunohistochemistry, ELISA, and Western Blot. For example, in some embodiments, the method for quantitatively detecting SCTAG at the protein level includes a specific antibody. Among them, a specific antibody refers to an antibody that can specifically recognize SCTAG protein, specifically including but not limited to monoclonal antibodies (such as full-length or complete monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (such as bispecific antibodies with expected biological activity), nano antibodies, or at least one of certain antibody fragments. In one or more embodiments of the present application, the test agent is an antibody against SCTAG protein; preferably, the antibody is a monoclonal antibody. When the expression of the SCTAG protein is positive and / or the expression of the SCTAG gene is positive, it indicates the risk of esophageal cancer (higher risk) or positive esophageal cancer.

[0084] Regarding the specific evaluation criteria of the present application, it can be understood that, generally, when the SCTAG test value is higher, it indicates that the subject tested has a higher probability of developing esophageal cancer; when the SCTAG test value is lower, it indicates that the subject tested has a lower probability of developing esophageal cancer.

[0085] In addition, according to the experimental data of the present application, the SCTAG can be used as an independent indication for the diagnosis or prediction of esophageal cancer, and an effective diagnosis of esophageal cancer can be achieved based only on SCTAG.

[0086] However, it is also understandable that in order to further enhance the diagnostic effect of the disease, the markers detected by the reagent, in addition to the SCTAG of the present application, may also include other nucleic acid fragments, proteins, metabolites, etc. with corresponding diagnostic capabilities that are already well known to those skilled in the art, and used as joint markers. In this way, the combined use of multiple biomarkers is achieved, and they cooperate with each other to achieve better diagnosis or prediction of esophageal cancer, thereby achieving a more effective evaluation purpose. Therefore, in addition to being an independent indication for the diagnosis of systemic lupus erythematosus, the SCTAG can also be combined with other existing known diagnostic markers to improve diagnostic efficiency.

[0087] 2. Products of this application

[0088] Based on the above uses, the detection agent or component capable of detecting SCTAG can be prepared into a corresponding diagnostic or predictive product, which can be used for the diagnosis or prediction of esophageal cancer.

[0089] The product includes reagents or components for detecting SCTAG markers. It is understood that such product forms are diverse, including but not limited to kit forms, system devices, and even computer readable media or computer system forms.

[0090] Kit Format

[0091] In some embodiments of the present application, a kit for detecting or analyzing SCTAG levels to predict esophageal cancer is also disclosed herein. Such a kit may include reagents for detecting the levels of one or more markers and instructions for predicting esophageal cancer based on the detected levels.

[0092] For example, the kits applied for can be of various types, such as a test kit for early warning of esophageal cancer; a test kit for early screening of esophageal cancer; a test kit for auxiliary diagnosis of esophageal cancer; a test kit for prognosis assessment of esophageal cancer; or a test kit for postoperative re-monitoring of esophageal cancer, etc. Regardless of the type of test kit, it contains the aforementioned core components: a detection agent or component capable of detecting SCTAG.

[0093] As mentioned above, the detection agent or component can qualitatively or quantitatively detect the SCTAG level, and the detection level includes the detection of nucleic acid level or protein level.

[0094] When the detection agent is detected at the nucleic acid level, it can be understood that there are many ways of nucleic acid detection in the art, including but not limited to sequencing technology, nucleic acid amplification technology, nucleic acid hybridization technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology, all of which can be used in this application. In some specific embodiments of the present application, including but not limited to any of the following specific methods: gene sequencing, polymerase chain reaction, isothermal amplification reaction, gene chip method, probe hybridization method, gel electrophoresis, Northern blotting, nucleic acid mass spectrometry or liquid chromatography. For example, according to some embodiments of the present application, the kit can be a PCR kit, a probe kit, a gene sequencing kit or a nucleic acid electrophoresis kit for SCTAG gene detection.

[0095] When the detection agent is detected at the protein level, it can be understood that there are many protein detection methods in the art, including but not limited to sequencing technology, immune technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology, all of which can be used in this application; in some specific embodiments of the present application, including but not limited to any of the following specific methods: amino acid sequencing, enzyme-linked immunosorbent assay, radioimmunoassay, immunohistochemistry, immunoblotting, flow cytometry, gel electrophoresis, protein spectrometry or liquid chromatography. For example, according to some embodiments of the present application, the kit can be an antibody kit for SCTAG protein detection (such as ELISA kit, WB kit, IHC kit, etc.), a protein sequencing kit, a protein gel electrophoresis kit or a chromatography kit, etc.

[0096] In some embodiments, such a kit may include a carrier, package or container that is compartmentalized to receive one or more containers, such as vials, tubes, etc., each of which contains one of the individual elements to be used in the method. The kit of the present application may include a container as described above and one or more other containers containing materials required from a commercial end-user standpoint, including buffers, diluents, filters, and package inserts with instructions for use.

[0097] In some embodiments, the kit further includes a sample processing reagent, and the sample processing reagent may include at least one of a sample lysis reagent, a sample purification reagent, and a sample extraction reagent.

[0098] In some embodiments, the product further comprises at least one of a standard, a calibrator, a reference substance, and a buffer. The reference substance is a control substance for testing the effectiveness of the experiment and serves as a control for judging the results. The buffer can be any solution known in the art that can provide appropriate buffer conditions during the detection process.

[0099] The kit can include the instructions for use of a group of reagents.For example, the kit can include instructions for performing at least one assay, such as immunoassay, protein binding assay, antibody-based assay, antigen-binding protein-based assay, protein-based array, enzyme-linked immunosorbent assay (ELISA), flow cytometry, protein array, blot, Western blot, nephelometry, turbidimetry, chromatography, mass spectrometry, enzyme activity and immunoassay, the immunoassay is selected from RIA, immunofluorescence, immunochemiluminescence, immunoelectrochemiluminescence, immunoelectrophoresis, competitive immunoassay and immunoprecipitation.In addition to the above-mentioned components, the kit will further include instructions for implementing the subject method.These instructions can be present in the subject kit in various forms, and one or more forms thereof can be present in the kit.A form in which these instructions can exist is that it is printed information on a suitable medium or substrate, for example, one or more sheets of printed information thereon, the paper is in the packaging of the kit, in the form of a package insert, etc.

[0100] Device form

[0101] In some embodiments of the present application, the present application also includes a device for detecting or analyzing SCTAG levels to predict esophageal cancer disease, which is configured to receive a mixture of one or more reagents and a test sample obtained from a subject to measure the SCTAG level, and a computer system that is communicatively coupled to the device to obtain the measured level and determine a score for predicting the likelihood of esophageal cancer disease in an esophageal cancer patient.

[0102] The device is configured to detect the SCTAG level in the mixture of reagent and test sample. For example, the device can determine the SCTAG level by immunoassay or the assay for nucleic acid detection. The mixture of reagent and test sample can be provided to the device by various containers, and the examples of these containers include holes, vials or tubes of orifice plates (for example, 96 orifice plates). Because of this, the device can have an opening (for example, a slot, a cavity, an opening, a sliding tray), and the opening can receive the container for accommodating the reagent test sample mixture and read to generate the quantitative expression value of a soluble medium. The example of the device includes an ELISA reader (for example, a luminescent ELISA reader, an absorbent ELISA reader, a fluorescent ELISA reader), a spectrometer and a spectrophotometer, and even some chip devices, etc.

[0103] Computer system or computer readable medium

[0104] In some embodiments of the present application, the present application also includes methods or uses of the present application that can be performed on a computer, including detecting or analyzing SCTAG levels by computer to predict esophageal cancer. For example, the construction and execution of a prediction model for generating a score can be implemented in hardware or software or a combination of the two. In one embodiment, a readable storage medium is provided, such as a medium including a data storage material encoded with machine-readable data, and when a machine programmed with instructions for using the data is used, the data storage material can display any data set and execution and results of the prediction model of the present application. Such data can be used for a variety of purposes, such as esophageal cancer patient monitoring, diagnosis, treatment considerations, etc. The embodiment of the above method can be implemented in a computer program executed on a programmable computer, the computer including a processor, a data storage system, a graphics adapter, a network adapter, at least one input device and at least one output device, etc. The display is coupled to the graphics adapter. The program code is applied to the input data to perform the above functions and generate output information. The output information is applied to one or more output devices in a known manner. The computer can be, for example, a personal computer, a microcomputer, or a workstation of conventional design.

[0105] 3. Diagnostic or predictive methods

[0106] The diagnostic or predictive methods of the present application include detection at different stages of the disease course. As mentioned above, the "diagnosis" of the present application may include early warning / prediction of the future onset of the disease, early screening or diagnosis, auxiliary diagnosis, diagnosis (determining whether the disease exists), prediction or judgment of the severity of the disease, prognosis assessment, monitoring treatment, such as monitoring disease deterioration or recurrence during treatment, evaluating the therapeutic efficacy of the disease, and the selection of a given treatment plan. Therefore, the diagnostic methods of the present application at least include esophageal cancer onset warning / prediction methods, esophageal cancer early screening / early diagnosis methods, esophageal cancer diagnosis / auxiliary diagnosis methods, esophageal cancer prognosis / postoperative recurrence assessment methods, etc.

[0107] Marker-based diagnostic methods, as known in the art, generally include the following basic steps or ideas:

[0108] (i) obtaining the level of SCTAG in the subject's sample;

[0109] (ii) comparing the SCTAG level with that of a control sample; wherein a significant difference in the SCTAG level between the subject sample and the control sample is an indication that the subject is at risk of developing esophageal cancer in the future; or,

[0110] (ii) comparing with a set threshold absolute amount; wherein the level of the subject's sample being higher than the threshold absolute amount is an indication that the subject has or will have esophageal cancer in the future.

[0111] The SCTAG level and the like are the same as described above.

[0112] In some specific embodiments of the present application, a set value of SCTAG level can be given, and the set value can be determined according to the expression level of SCTAG of normal samples of normal people and / or non-esophageal cancer patients. For example, the average value of the expression level of SCTAG of normal samples of an appropriate number of samples is selected, or a reasonable multiple is set based on the average value, such as 0.9 times, 0.8 times, 0.7 times, 0.6 times, 0.5 times, etc. When the expression level of SCTAG of the subject is higher than the set value, it is judged to be esophageal cancer. It can be understood that the set value determined based on the average value, or the multiple of the average value needs to have good classification significance, and the known samples can be tested by using commonly used statistical test methods through the classification based on the set value. When the result is statistically significant, it indicates that the set value can be used as a judgment standard.

[0113] As used herein, the terms "individual," "subject," and "patient" are used interchangeably herein and refer to any mammalian subject, particularly a human, in need of diagnosis, treatment or therapy.

[0114] As used herein, the terms "sample", "specimen", "test sample", "subject sample", etc. include various sample types obtained from patients, individuals or subjects and can be used for diagnosis or monitoring determination. Patient samples can be obtained from healthy subjects, sick patients or patients with symptoms related to esophageal cancer. In addition, the sample obtained from the patient can be divided into parts, and only a part can be used for diagnosis. In addition, the sample or a part thereof can be stored under the condition of maintaining the sample for subsequent analysis. This definition specifically includes blood and other liquid samples of biological origin (including but not limited to tissues, cells (including cell culture supernatants), body fluids, serum, plasma, whole blood, urine, semen, saliva, pleural effusion, ascites and feces, etc.). In a specific embodiment, the sample includes a blood sample. In another embodiment, a serum sample is used. This definition also includes samples that are manipulated in any way after the sample is obtained, such as by centrifugation, filtration, precipitation, dialysis, chromatography, treatment with reagents, washing or enrichment of certain cell populations. These terms also include clinical samples, and also include cells in culture, cell supernatants, tissue samples, organs, etc. Samples may also include fresh frozen and / or formalin-fixed, paraffin-embedded tissue blocks, such as blocks prepared from clinical or pathological biopsies, prepared for pathological analysis or studies by immunohistochemistry.

[0115] It is understandable that the subject sample or the control sample can be selected according to actual needs. For example, in the prediction / warning of esophageal cancer, or early screening / diagnosis of esophageal cancer, the subject sample is the sample to be checked, and the control sample is a healthy sample; in one or more embodiments of the present application, when the SCTAG detection value is high, it indicates that the subject has a higher chance of developing esophageal cancer; when the SCTAG detection value is low or absent, it indicates that the subject has a lower chance of developing esophageal cancer. When esophageal cancer is diagnosed or assisted in diagnosis, the subject sample is a sample from a suspected patient, and the control sample is a healthy sample.

[0116] In the method for evaluating the prognosis / recurrence of esophageal cancer, the subject sample is a sample of an esophageal cancer patient after treatment, and the control sample is a sample of an esophageal cancer patient before treatment; in some specific embodiments, when the prediction or diagnosis is for postoperative recurrence monitoring, the subject sample is a sample of an esophageal cancer patient who has received postoperative treatment, and the control sample is a sample of an esophageal cancer patient before preoperative treatment. The specific method includes the following steps: (i) obtaining the level of SCTAG in the postoperative sample of the subject; (ii) comparing the SCTAG level with the preoperative control sample of the subject; when the SCTAG level in the postoperative sample increases or remains unchanged, it indicates that the risk of recurrence of esophageal cancer is high; when the SCTAG level in the postoperative sample decreases, it indicates that the probability of recurrence of postoperative esophageal cancer in the subject is low.

[0117] The implementation scheme of the present application is described in detail below with reference to examples.

[0118] Sample and clinical data collection involved in this embodiment

[0119] Tissue: Tissue samples from 90 patients with esophageal cancer were obtained from the tissue specimen bank of the Third Hospital of Jilin University and the First Hospital of Jilin University. The tissue samples of each patient included tumor tissue, adjacent tissue (2-5 cm away from the tumor), and normal tissue (>5 cm away from the tumor) for extraction of total protein and mRNA.

[0120] Serum: The tissue sample bank of Jilin University Third Hospital and Jilin University First Hospital obtained preoperative serum samples from 244 patients with esophageal cancer and serum samples from 220 healthy subjects. In addition, serum samples were collected from 59 patients undergoing esophageal cancer surgery before and on days 3-8 after surgery. All sera were collected from venous blood in the early morning in an empty stomach using EDTA anticoagulation tubes. The serum collected in this part was used for enzyme-linked immunosorbent assay.

[0121] Pathological sections: Pathological sections of tumor tissues from 68 esophageal cancer patients, as well as corresponding paracancerous tissues and normal esophageal tissues were obtained from the Department of Pathology of the Third Hospital of Jilin University.

[0122] At the same time, their relevant clinical pathological data were collected, including age, gender, tumor location, tumor gross classification, tumor differentiation, depth of invasion, presence of lymph node metastasis, presence of vascular and neural invasion, 5-year survival rate, short-term recurrence rate of esophageal cancer (recurrence criteria: non-surgical hoarseness, mediastinal lymphadenopathy, distant metastasis, etc. within 1 year), and the expression of conventional tumor markers such as CEA, AFP, CA199, and CA724. The TNM staging of esophageal cancer was determined according to the 8th edition of the UICC / AJCC TNM staging guidelines for esophageal cancer.

[0123] Example 1: RT-PCR detection of SCTAG gene levels in esophageal cancer tissues

[0124] The applicant collected tissue samples from 50 patients with esophageal cancer. Each patient included esophageal cancer tissue, paracancerous tissue (2cm-5cm away from the edge of esophageal cancer), and normal tissue (more than 5cm away from the edge of esophageal cancer).

[0125] Agarose gel electrophoresis was performed on the qRT-PCR products of tumor tissues and normal tissues, and the results showed that different levels of SCTAG mRNA expression were present in esophageal cancer tissues. Figure 1 A; however, there is no corresponding SCTAG band at the same position of the qRT-PCR product in normal tissues, see Figure 1 B (The faint bands observed below 100 bp of DNA marker are primer dimers. Figure 1 Also present below the positive band in A).

[0126] Further quantitative analysis of SCTAG mRNA was performed on each sample (GAPDH was used as an internal reference). The results showed that compared with the paired normal tissues, SCTAG mRNA expression was upregulated in 64% of esophageal cancer tissues, see Figure 2 A, SCTAG mRNA expression was upregulated in 66% of esophageal adjacent tissues, see Figure 2 B, It can be seen that there are significant differences in genes between esophageal cancer tissue and normal tissue.

[0127] Example 2: Immunohistochemical detection of SCTAG protein expression in esophageal cancer tissues

[0128] In this example, 68 paraffin tissue sections of esophageal cancer were collected, and paracancerous tissue and normal esophageal mucosal tissue from the same patient were used as controls.

[0129] Immunohistochemical staining showed that SCTAG protein was widely expressed in esophageal cancer tissues, and positive staining of SCTAG protein was observed in the cytoplasm of esophageal cancer cells. SCTAG was also expressed in paracancerous tissues of the esophageal cancer, but the expression intensity was significantly lower than that in cancer tissues, while SCTAG was rarely or not expressed in normal tissues. Figure 3 shown.

[0130] SCTAG has the highest expression level in esophageal cancer tissue and is highly specific. This result shows that SCTAG can be used for the diagnosis of esophageal cancer and has a certain early warning effect on the occurrence of esophageal cancer.

[0131] Example 3: Western-Blot detection of SCTAG protein expression in esophageal cancer tissue

[0132] In this example, fresh esophageal cancer tissues were collected, and normal esophageal mucosal tissues from the same patient were used as controls.

[0133] Western-Blot showed that the expression of SCTAG protein in esophageal cancer tissue was significantly higher than that in the corresponding normal tissue. In normal esophageal cancer mucosal tissue, SCTAG was also expressed in some normal tissues, but the expression intensity was significantly lower than that in cancer tissues, such as Figure 4 shown.

[0134] This result shows that SCTAG can be used for the diagnosis of esophageal cancer and has a certain early warning effect on the occurrence of esophageal cancer.

[0135] Example 4. Diagnostic value of serum SCTAG levels in patients with esophageal cancer

[0136] In this example, the concentration of SCTAG protein in the peripheral serum of healthy subjects and esophageal cancer patients before surgery was detected by enzyme-linked immunosorbent assay to analyze the diagnostic ability of serum SCTAG level for esophageal cancer.

[0137] The results showed that the serum SCTAG level in early esophageal cancer was significantly higher than that in healthy subjects (P<0.001, Figure 5 A), with the progression of tumor, serum SCTAG protein level also increased significantly (P<0.001, Figure 5 A). The diagnostic ability of SCTAG protein in stage I esophageal cancer is high (AUC = 0.9913, Figure 5 B), in addition, SCTAG has high diagnostic sensitivity and specificity in all stages of esophageal cancer, as shown in Table 1. In addition, this example also compares SCTAG with common tumor markers (CEA, AFP, CA199, CA724), and the results show that SCTAG is significantly better than existing tumor markers ( Figure 5 C).

[0138] Table 1. Diagnostic sensitivity and specificity of SCTAG in various stages of esophageal cancer

[0139]

[0140] The experimental results showed that SCTAG expression was significantly elevated in the serum of patients with esophageal cancer and could be used as a marker for early warning and diagnosis of esophageal cancer.

[0141] Example 5: Serum SCTAG level indicates prognosis, recurrence and prognosis assessment of esophageal cancer

[0142] In this example, preoperative and postoperative serum samples were collected from 59 patients with esophageal cancer who underwent surgery at the First Hospital of Jilin University, and the changes in SCTAG protein concentration in peripheral serum before and after surgery were detected by enzyme-linked immunosorbent assay to evaluate the correlation between changes in serum SCTAG levels and postoperative recurrence of esophageal cancer.

[0143] The results showed that compared with the preoperative serum SCTAG level, approximately 55.9% (33 / 59) of the patients had a decrease in serum SCTAG after surgery, approximately 42.4% (25 / 59) of the patients had an increase in serum SCTAG after surgery, and one patient's preoperative and postoperative serum SCTAG levels remained unchanged ( Figure 6 A). The patients were then followed up 1 year after surgery. Statistics showed that only 6% (2 / 33) of the patients with decreased serum SCTAG after surgery experienced recurrence, while as many as 44% (11 / 25) of the patients with increased serum SCTAG after surgery experienced recurrence ( Figure 6 B), and one patient whose serum SCTAG level remained unchanged also relapsed within one year.

[0144] In addition, this study also conducted a Kaplan–Meier analysis of the SCTAG expression level in esophageal cancer tissue and the 5-year survival of patients. The results showed that the 5-year survival rate of patients with higher SCTAG levels in tumor tissue was lower than that of patients with lower SCTAG levels, indicating that the prognosis of patients was poor ( Figure 6 C).

[0145] In addition, in the group of patients with increased SCTAG after surgery, the symptoms of recurrence appeared earlier, and they were more likely to have distant metastasis rather than local lymph node metastasis when they relapsed ( Figure 7 ).

[0146] The results of the above examples show that serum SCTAG levels are significantly increased in patients with esophageal cancer, have a strong ability to diagnose esophageal cancer, and can better assess the risk of esophageal cancer; the increase in serum SCTAG before and after surgery indicates that the risk of recurrence of esophageal cancer is also increased.

[0147] The foregoing description of specific exemplary embodiments of the present application is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present application to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present application and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present application and various different options and changes. The scope of the present application is intended to be limited by the claims and their equivalents.

Claims

1. Use of SCTAG protein or gene as a marker in the diagnosis or prediction of esophageal cancer, or in the preparation of products for the diagnosis or prediction of esophageal cancer.

2. Use of a detection agent or component for obtaining the SCTAG level in a sample in the diagnosis or prediction of esophageal cancer, or in the preparation of a product for the diagnosis or prediction of esophageal cancer.

3. A product for diagnosing or predicting esophageal cancer, characterized in that: Contains a detection agent or component for obtaining the level of SCTAG in a sample.

4. According to claim 1 or 2 or 3, it is characterized in that The diagnosis or prediction particularly includes the onset warning, early diagnosis, prognosis assessment or recurrence warning of esophageal cancer.

5. According to claim 1 or 2 or 3, it is characterized in that The sample comprises one or more of tissue, serum, plasma, whole blood, urine, saliva, pleural effusion, ascites or cell culture supernatant; preferably, the sample comprises tissue, serum, plasma or whole blood.

6. According to claim 1 or 2 or 3, it is characterized in that The level includes gene level or protein level.

7. According to claim 6, it is characterized in that The nucleic acid level is obtained by sequencing technology, nucleic acid amplification technology, nucleic acid hybridization technology, electrophoresis technology, biological mass spectrometry technology or chromatography technology; preferably, the nucleic acid level acquisition method includes but is not limited to any one of the following methods: gene sequencing method, polymerase chain reaction method, isothermal amplification reaction method, gene chip method, probe hybridization method, gel electrophoresis method, Northern blotting method, nucleic acid mass spectrometry or liquid chromatography.

8. According to claim 6, it is characterized in that The protein level is obtained by sequencing technology, immunoassay technology, electrophoresis technology, biomass spectrometry technology or chromatography technology; preferably, the protein level acquisition method includes but is not limited to any of the following methods: amino acid sequencing, enzyme-linked immunosorbent assay, chemiluminescence, immunochromatography, radioimmunoassay, immunohistochemistry, immunoblotting, flow cytometry, gel electrophoresis, protein spectrometry or liquid chromatography.

9. According to claim 8, it is characterized in that The protein level is obtained by immunoassay, wherein the detection agent is a reagent for detecting the expression of SCTAG protein; preferably, the reagent is an antibody against SCTAG protein; more preferably, the antibody against SCTAG protein is a monoclonal antibody.

10. According to claim 1 or 2 or 3, it is characterized in that The product comprises a kit or a device.

11. According to claim 10, it is characterized in that The kit also includes sample processing reagents, which include: at least one or more of a sample lysis reagent, a sample purification reagent, and a sample nucleic acid or protein extraction reagent.

12. A method for early diagnosis or early warning of esophageal cancer, characterized in that: The steps include: (i) obtaining the level of SCTAG in the subject's sample; (ii) comparing the SCTAG level with that of a control sample; wherein a significant difference in the SCTAG level between the subject sample and the control sample is an indication that the subject has or will have esophageal cancer in the future; or, (ii) comparing with a set threshold absolute amount; wherein the level of the subject's sample being higher than the threshold absolute amount is an indication that the subject has or will have esophageal cancer in the future.

13. A method for early warning and assessment of postoperative recurrence of esophageal cancer, characterized in that: The steps include: (i) obtaining the level of SCTAG in the postoperative samples of the subjects; (ii) comparing the SCTAG level with that of the subject's preoperative control sample; when the SCTAG level in the postoperative sample increases or remains unchanged, it indicates a high risk of esophageal cancer recurrence.

14. A method for detecting a marker in a subject having or suspected of having esophageal cancer in vivo or in vitro, characterized in that: The method comprises the step of determining or detecting the level of SCTAG in a sample from a subject.

15. A method for evaluating or screening diagnostic or predictive markers for esophageal cancer, characterized in that: The process includes the steps of correlation or consistency analysis between potential biomarkers and SCTAG.

16. A method for in vivo or in vitro screening of therapeutic agents for esophageal cancer, characterized in that: The method includes the step of evaluating the level of SCTAG in a sample treated with a therapeutic agent.

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