CN116699134A - Application of SCTAG2 in gastric cancer diagnosis and targeting drugs - Google Patents

Application of SCTAG2 in gastric cancer diagnosis and targeting drugs Download PDF

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CN116699134A
CN116699134A CN202210186194.3A CN202210186194A CN116699134A CN 116699134 A CN116699134 A CN 116699134A CN 202210186194 A CN202210186194 A CN 202210186194A CN 116699134 A CN116699134 A CN 116699134A
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sctag2
protein
nucleic acid
expression
gastric cancer
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陈芳芳
房学东
任圣男
杨丁全
邢磊
董永立
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Abstract

The invention belongs to the field of biological medicine, relates to novel application of SCTAG2, and in particular relates to application of an SCTAG2 protein or nucleic acid for encoding the SCTAG2 protein as a diagnosis marker in preparation of a gastric cancer diagnosis or auxiliary diagnosis detection agent, and provides that the expression level of the SCTAG2 in gastric cancer tissues is abnormal for the first time, and the expression level of the SCTAG2 in gastric cancer tissues with larger volume is higher when clinical stage is later. Since tumor volume and clinical stage are risk factors affecting tumor prognosis, it is suggested that the higher the expression level of SCTAG2, the worse the prognosis of gastric cancer patients.

Description

Application of SCTAG2 in gastric cancer diagnosis and targeting drugs
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to application of SCTAG2 in gastric cancer diagnosis and targeted therapy, which comprises application in gastric cancer screening, gastric cancer diagnosis, gastric cancer monitoring, gastric cancer medication guidance and/or gastric cancer prognosis judgment.
Background
At present, worldwide, the death rate of gastric cancer is the second most, the median survival time of gastric cancer in the progressive stage is only 12 months, and about 783000 patients die from gastric cancer worldwide each year. Gastric cancer is a multifactorial disease that includes genetic and environmental factors such as eating habits, smoking, and helicobacter pylori infection. The incidence of gastric cancer has obvious gender and region differences, and the risk of occurrence of gastric cancer of men is two to three times higher than that of women, and meanwhile, compared with developed countries, the incidence rate of gastric cancer in developing countries is also higher, such as China, south America, eastern Europe and east Asia countries. Meanwhile, prognosis of gastric cancer patients is poor, and according to statistics, the survival rate of gastric cancer patients in China is 36% in 5 years, and the survival rates of five years in japan and korea are both over 60%. The main reason is that the early diagnosis rate is low and the urban and rural medical and hygienic resource allocation is unbalanced. Due to lack of physical examination consciousness and insufficient popularity of gastroscopy screening, gastric cancer patients in China have later stage in clinical diagnosis, and lymph node or blood metastasis appears, so that the treatment effect is poor. Therefore, early diagnosis and early treatment are the key for improving the survival rate of gastric cancer patients, and the early diagnosis and early treatment are used for reliably early warning in the early stage of disease occurrence by developing a novel gastric cancer diagnosis technology, so that the screening of high-risk groups is facilitated, the early diagnosis level of gastric cancer is improved, and the effective prevention and control of gastric cancer are realized.
The diagnosis of gastric cancer mainly depends on digestive tract endoscopy, imaging examination, tumor marker detection and the like, wherein the tumor marker detection has the advantages of low cost, small wound and easiness in repetition, and has wider application space. Tumor markers refer to substances that are characteristic of being present in malignant tumor cells, or produced abnormally by tumor cells, or produced by the body's stimulatory response to a tumor. These tumor markers are generally present in the form of metabolites such as antigens, enzymes, hormones, etc. in the tissues, blood and secretions of tumor patients and can be detected by immunological and biochemical methods, etc. The detection of the tumor markers can be helpful for judging the occurrence and the type of the tumor, can well evaluate the stage and the treatment effect of the cancer, and can monitor the recurrence and the metastasis of the tumor.
Carcinoembryonic antigen (CEA) was originally discovered in 1965, and was first used in the diagnosis of gastric cancer in 1981, and can effectively identify and screen high risk populations of gastric cancer. However, serum CEA concentrations can be elevated in chronic diseases such as colitis and other types of cancer and thus are a less specific tumor marker with lower tumor localization. But serum CEA concentration detection can monitor tumor recurrence, and can also be combined with other indexes to evaluate the chemotherapy effect of gastric cancer. CA19-9, CA72-4, CA50 and the like can be used for diagnosing gastric cancer, but the sensitivity and the specificity are not ideal. At present, CEA is still considered as one of the most valuable serum markers for gastric cancer diagnosis, and is often clinically detected in combination with tumor markers such as CA72-4 and the like for improving sensitivity and accuracy. SCTAG2 belongs to the cancer testis antigen family (Cancer testis antigen, CTA), is abnormally expressed only in male testis tissue and malignant tumor tissue, has high tissue specificity, and is an ideal tumor diagnosis marker.
The occurrence of gastric cancer is often considered to be the result of a multifactorial process, including host reactions, bacterial virulence, diet and other environmental factors, and the like. The incidence rate of gastric cancer can be obviously reduced by properly managing and protecting the dangerous factors. Therefore, development of a novel detection agent and kit for diagnosis of gastric cancer, and a drug for treatment and prevention of gastric cancer have been demanded.
Disclosure of Invention
The invention provides that the expression level of the SCTAG2 in gastric cancer tissues is abnormal, and the expression level of the SCTAG2 in gastric cancer tissues with later clinical stage and larger volume is higher. Since tumor volume and clinical stage are risk factors affecting tumor prognosis, it is suggested that the higher the expression level of SCTAG2, the worse the prognosis of gastric cancer patients.
The invention provides application of an SCTAG2 protein or a nucleic acid encoding the SCTAG2 protein as a diagnosis marker in preparation of a gastric cancer diagnosis or auxiliary diagnosis detection agent.
In one or more embodiments of the invention, the detection agent is an agent that detects the expression of an SCTAG2 protein, the agent being an antibody to the SCTAG2 protein; more preferably, the antibody against SCTAG2 protein is a monoclonal antibody.
In one or more embodiments of the invention, the detection agent is an expression agent that detects a nucleic acid encoding an SCTAG2 protein, the agent being a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out a nucleic acid encoding an SCTAG2 protein; more preferably, the polynucleotide is an siRNA or shRNA, or a guide rna for a CRISPR/Cas9 system.
In one or more embodiments of the invention, the detection is a quantitative or qualitative detection. The detection of SCTAG2 protein expression refers to detection of whether SCTAG2 protein expression is positive or negative. Detecting expression of a nucleic acid encoding an SCTAG2 protein refers to detecting whether expression of the nucleic acid encoding an SCTAG2 protein is positive or negative.
In one or more embodiments of the invention, when the expression of the SCTAG2 protein is detected as positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is detected as positive: prompting the risk (higher risk) or positive of gastric cancer; or to indicate a pre-cancerous pre-warning risk of gastric cancer, for example to indicate a risk of progression of a pre-cancerous lesion of gastric cancer (higher risk). The type of the conclusion can be judged by combining the self situation of the object, and the self situation can be: subjects were at a stage of premalignant lesions (basal cell hyperplasia, mild dysplasia, moderate dysplasia, severe dysplasia or carcinoma in situ), subjects were highly suspected of having gastric cancer, gastroscopic results of subjects, etc. The occurrence and development of gastric cancer are 5 stages according to the histological sequence: basal cell proliferation, mild dysplasia, moderate dysplasia, severe dysplasia and carcinoma in situ.
In one or more embodiments of the present invention, the progression of the premalignant lesion is from a relatively anterior stage to a later stage.
In one or more embodiments of the present invention, the sample used is a tissue sample of the subject to be tested; preferably, it is a surgical tissue sample or a biopsy sample, such as an endoscopic biopsy sample.
In one or more embodiments of the invention, the use is the use of preparing a detector for diagnosis of gastric cancer, wherein the risk (higher risk) or positivity of gastric cancer is indicated when the expression of the SCTAG2 protein is positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is positive. Preferably, further screening such as histopathological examination is performed.
In one or more embodiments of the invention, the use is the use of the preparation of a detector for the prediction of the risk of progression of a gastric precancerous lesion, wherein the expression of the SCTAG2 protein is positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is positive, is indicative of a risk of progression of a gastric precancerous lesion (higher risk). Preferably, further screening such as histopathological examination is performed.
The detection agent is used to perform one of polymerase chain reaction or denaturing gradient gel electrophoresis or nucleic acid sequencing or nucleic acid separation chip detection or denaturing high performance liquid chromatography or in situ hybridization or bio-mass spectrometry or HRM methods when the agent is at the nucleic acid level;
the detection agent is used to perform one of a biological mass spectrometry or an amino acid sequencing or electrophoresis method when the agent is at the protein level.
The invention also provides a detection kit for screening gastric cancer.
The invention also provides a detection kit for diagnosing gastric cancer.
The invention also provides a detection kit for monitoring gastric cancer.
The invention also provides a detection kit for guiding the administration of gastric cancer and/or prognosis of gastric cancer.
In one or more embodiments of the invention, the detection kit comprises one of the following detection agents: (1) an agent that detects the expression of SCTAG2 protein; (2) An agent that detects expression of a nucleic acid encoding an SCTAG2 protein.
In one or more embodiments of the invention, the kit includes reagents for detecting other pre-warning markers associated with risk of gastric precancerous lesions in addition to the reagents (1) and (2) above, preferably, the other reagents include, but are not limited to, protamine antigen 1 (SCTAG 2), carbohydrate antigen 72-4 (CA 72-4), carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 24-2 (CA 24-2), carbohydrate antigen 50 (CA 50), carcinoembryonic antigen (CEA), gastrin 17 (G-17).
In one or more embodiments of the invention, the kit further comprises a sample processing reagent comprising at least one of a sample lysing reagent, a sample purifying reagent, and a sample nucleic acid or protein extracting reagent.
The present invention also provides a use of any one selected from the following items (1) to (4) in the preparation of a medicament for the treatment or co-treatment or prevention of gastric cancer: (1) SCTAG2 protein; (2) a nucleic acid encoding an SCTAG2 protein; (3) An agent that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein; (4) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein.
In one or more embodiments of the invention, the agent that reduces the level of SCTAG2 protein expression or inhibits the expression of SCTAG2 protein in item (3) is an antibody against SCTAG2 protein; preferably, the antibody against SCTAG2 protein is a monoclonal antibody;
in one or more embodiments of the present invention, the agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein in item (4) is a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out a nucleic acid encoding an SCTAG2 protein; preferably, the polynucleotide is an siRNA or shRNA, or a guide rna for a CRISPR/Cas9 system.
The present invention also provides a medicament comprising the following item (1) or item (2), (1) a medicament that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein; (2) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein.
In one or more embodiments of the invention, the agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein is a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out a nucleic acid encoding an SCTAG2 protein; preferably, the polynucleotide is an siRNA or shRNA, or a guide rna for a CRISPR/Cas9 system.
In one or more embodiments of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.
The present invention also provides a pharmaceutical product comprising an effective amount of the following item (1) or item (2);
(1) A drug that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein;
(2) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein.
In one or more embodiments of the invention, the first product and the second product each independently comprise one or more pharmaceutically acceptable excipients.
In one or more embodiments of the invention, the amino acid sequence of the SCTAG2 protein is shown in SEQ ID NO. 1.
In one or more embodiments of the invention, the sequence of the nucleic acid encoding the SCTAG2 protein is shown in SEQ ID NO. 2.
In one or more embodiments of the invention, the gastric cancer includes, but is not limited to, gastric adenocarcinoma, adenosquamous carcinoma, squamous carcinoma, and carcinoid.
The beneficial effects are as follows:
the invention discovers and proves that the SCTAG2 can be used for screening gastric cancer, diagnosing gastric cancer, monitoring gastric cancer, guiding gastric cancer medication and/or prognosis of gastric cancer for the first time.
Firstly, an application of a reagent for detecting the content of SCTAG2 in a kit for detecting gastric cancer is provided. The gastric cancer detection comprises gastric cancer screening, gastric cancer diagnosis, gastric cancer monitoring, gastric cancer medication guidance and/or gastric cancer prognosis. The detection is performed on at least one of lavage fluid, tissue or tissue lysate, cell lysate, blood, serum, plasma of the subject. The SCTAG2 content is nucleic acid content or protein content. When the reagent is at the nucleic acid level, the detection reagent is used to perform any one of the following methods: polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid sequencing, nucleic acid separation chip detection, denaturing high performance liquid chromatography, in situ hybridization, biological mass spectrometry, and HRM methods. When the agent is at the protein level, the detection agent is used to perform any one of the following methods: biological mass spectrometry, amino acid sequencing, electrophoresis, and detection with antibodies specifically designed for the mutation site.
Also provided is a gastric cancer detection method for detecting the SCTAG2 content in a sample against a subject sample in vivo or in vitro;
there is also provided a gastric cancer detection model or evaluation system, characterized in that the model comprises: a detection module that enables detection of SCTAG2 content in a sample against a subject sample, either in vivo or in vitro; and the analysis module is used for carrying out analysis and evaluation according to the result of the detection module.
Also provided is a use of any one selected from the following items (1) to (4) in the manufacture of a medicament for the treatment or co-treatment or prevention of gastric cancer: (1) SCTAG2 protein; (2) a nucleic acid encoding an SCTAG2 protein; (3) An agent that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein; (4) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein.
The subject sample is not limited, so that the probability of diagnosing gastric cancer by the subject can be clarified, and the prognosis condition of a gastric cancer patient can be evaluated in a related manner.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 expression levels of SCTAG2 in gastric cancer tissue and paracancerous normal tissue; detecting the expression condition of the SCTAG2 protein in tissues through an immunohistochemical experiment; a-c are stomach adenocarcinoma tissues, d-f are normal gastric mucosa tissues; a, d are amplified 10 times, b, e are amplified 20 times, c, f are amplified 40 times.
FIG. 2 real-time fluorescent quantitative PCR detection of the expression level of SCTAG2 in gastric cancer tissue, paracancerous tissue and normal tissue.
Detailed Description
The following description of the embodiments of the present invention will be made apparent and fully in view of the accompanying drawings, in which some, but not all embodiments of the invention are shown. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The following terms or definitions are provided solely to aid in the understanding of the invention. These definitions should not be construed to have a scope less than understood by those skilled in the art.
Unless defined otherwise hereinafter, all technical and scientific terms used in the detailed description of the invention are intended to be identical to what is commonly understood by one of ordinary skill in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.
As used herein, the terms "comprising," "including," "having," "containing," or "involving" are inclusive or open-ended and do not exclude additional unrecited elements or method steps. The term "consisting of …" is considered to be a preferred embodiment of the term "comprising". If a certain group is defined below to contain at least a certain number of embodiments, this should also be understood to disclose a group that preferably consists of only these embodiments.
The indefinite or definite article "a" or "an" when used in reference to a singular noun includes a plural of that noun.
The terms "about" and "substantially" in this invention mean the range of accuracy that one skilled in the art can understand yet still guarantee the technical effect of the features in question. The term generally means a deviation of + -10%, preferably + -5%, from the indicated value.
Furthermore, the terms first, second, third, (a), (b), (c), and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.
The diagnosis is based on the SCTAG2 as an oncogene, abnormal expression occurs in the stomach cancer forming process, and the gene has important biological functions, so that the SCTAG2 can be used for stomach cancer diagnosis and prognosis evaluation research. Accordingly, the gastric cancer detection methods provided by the present invention in some aspects are one such method: it is contemplated that it is within the scope of the invention to include an SCTAG2 content assay step for gastric cancer detection in a subject.
The gastric cancer detection of the invention comprises, but is not limited to, gastric cancer screening, gastric cancer diagnosis, gastric cancer monitoring, gastric cancer medication guidance, gastric cancer prognosis judgment and/or the like.
The converted protein SCTAG 2) belongs to cancer testis antigen family (Cancer testis antigen, CTA), is abnormally expressed only in male testis tissue and malignant tumor tissue, has high tissue specificity, and is an ideal tumor diagnosis marker. Without limitation, the amino acid sequence of the SCTAG2 is as shown in the prior art, such as SEQ ID NO. 1:
MSTSRKLKSHGMRRSKSRSPHKGVKRGGSKRKYRKGNLKSRKRGD DANRNYRSHL；
the corresponding coding nucleic acid sequences are also known in the art, for example as shown in SEQ ID NO. 2:
atgtcgacca gccgcaaatt aaagagtcat ggcatgagga ggagcaagag ccgatctcct cacaagggag tcaagagagg tggcagcaaa agaaaatacc gtaagggcaa cctgaaaagt aggaaacggg gcgatgacgc caatcgcaat taccgctccc acttgtga。
the analysis steps of the present invention are not limited to the type of sample, the state of the sample, the processing of the sample, or the detection means.
For example, in some embodiments, the sample type may be at least one selected from the group consisting of blood, serum, plasma, urine, lavage, tissue or tissue lysates, cell culture supernatants of the subject;
in some embodiments, the sample state may be in vivo or in vitro; whatever pretreatment the sample is subjected to, it is within the scope of the invention as long as the final purpose is to perform the SCTAG2 content detection on the sample.
Of course, the sample detection means of the present invention may be varied and any currently known means available for quantitative or qualitative determination of proteins or genes are theoretically suitable for use in the present invention, and thus, in some embodiments, the SCTAG2 level is nucleic acid content or protein content.
In some embodiments, the detection agent detects at the nucleic acid level.
The nucleic acid level (DNA or RNA level) detecting agent may be any agent known to those skilled in the art, for example, a nucleic acid (usually a probe or primer) which hybridizes to the DNA or RNA and is labeled with a fluorescent label. And it is also easy for those skilled in the art to detect cDNA after reverse transcription of mRNA into cDNA, conventional substitutions of these techniques do not exceed the scope of the present invention.
In some embodiments, the detection agent is used to perform any one of the following methods:
polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid sequencing, nucleic acid separation chip detection, denaturing high performance liquid chromatography, in situ hybridization, biological mass spectrometry, and HRM methods. In some embodiments, the polymerase chain reaction is selected from the group consisting of restriction fragment length polymorphism method, single strand conformational polymorphism method, taqman probe method, competitive allele-specific PCR, and allele-specific PCR.
In some embodiments of the invention, the nucleic acid sequencing method may be transcriptome sequencing or genomic sequencing. In still other embodiments of the invention, the nucleic acid sequencing method is high throughput sequencing, also known as second generation sequencing ("NGS"). NGS is distinguished from "Sanger sequencing" (first generation sequencing), which is based on the electrophoretic separation of chain termination products in a single sequencing reaction. NGS is a revolutionary revolution to traditional Sanger sequencing technology, and can sequence hundreds of thousands to millions of nucleic acid molecules at a time. Sequencing platforms that can use NGS of the present invention are commercially available, including but not limited to Roche/454FLX, illumina/Solexa GenomeAnalyzer and Applied Biosystems SOLID system, and the like. Transcriptome sequencing can also rapidly and comprehensively obtain almost all transcripts and gene sequences of specific cells or tissues of a certain species under a certain state through a second generation sequencing platform, and can be used for researching gene expression quantity, gene functions, structure, alternative splicing, new transcript prediction and the like. In other embodiments of the invention, the nucleic acid sequencing method may be single molecule real-time sequencing, and single molecule DNA sequencing technology is a new generation sequencing technology developed in recent 10 years, also called third generation sequencing technology, including single molecule real-time sequencing, true single molecule sequencing, single molecule nanopore sequencing, and the like.
In some embodiments, the detection agent detects at the protein level.
In some embodiments, the detection agent is used to perform any one of the following methods: biological mass spectrometry, amino acid sequencing, electrophoresis, and detection with antibodies specifically designed for the mutation site. The detection method using an antibody specifically designed for the mutation site may further be immunoprecipitation, co-immunoprecipitation, immunohistochemistry, ELISA, western Blot, etc.
It will be appreciated that with respect to the specific evaluation criteria of the present invention, when SCTAG2 is expressed positively, the higher the probability of diagnosing gastric cancer in the subject is suggested, and the higher the expression level, the worse the prognosis is suggested; when SCTAG2 was expressed negatively, the subjects were prompted to have a lower chance of diagnosing gastric cancer.
Of course, the evaluation application of the invention can be not only single detection of SCTAG2, but also detection in combination with other markers known at present, such as: protamine antigen 1 (SCTAG 2), pepsinogen I/II, carbohydrate antigen 72-4 (CA 72-4), carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 24-2 (CA 24-2), carbohydrate antigen 50 (CA 50), carcinoembryonic antigen (CEA), and gastrin 17 (G-17).
Likewise, the application of the reagent for detecting the SCTAG2 content in the gastric cancer detection kit is not limited by the sample type, the sample state, the sample treatment, the detection means and the like; the gastric cancer detection comprises gastric cancer screening, gastric cancer diagnosis, gastric cancer monitoring, gastric cancer medication guidance, gastric cancer prognosis judgment and/or the like.
In some embodiments, the detection is performed against at least one of blood, serum, plasma, tissue or tissue lysate, cell culture supernatant of the subject.
In some embodiments, the SCTAG2 content is a nucleic acid content or a protein content. Further, when the reagent is at the nucleic acid level, the detection reagent is used to perform any one of the following methods: polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid sequencing, nucleic acid separation chip detection, denaturing high performance liquid chromatography, in situ hybridization, biological mass spectrometry, and HRM methods. Further, when the agent is at the protein level, the detection agent is used to perform any one of the following methods: biological mass spectrometry, amino acid sequencing, electrophoresis, and detection with antibodies specifically designed for the mutation site.
Regarding the specific evaluation criteria of the present invention, it can be appreciated that when SCTAG2 is expressed positively, the higher the probability of diagnosing gastric cancer by the subject is prompted, and the higher the expression level, the worse the prognosis is prompted; when SCTAG2 was expressed negatively, the subjects were prompted to have a lower chance of diagnosing gastric cancer.
Of course, for the purpose of detecting comprehensiveness or more accuracy of results, the evaluation application of the present invention may be not only a single detection for SCTAG2, but also a detection in combination with a currently known marker, such as: protamine antigen 1 (SCTAG 2), pepsinogen I/II, carbohydrate antigen 72-4 (CA 72-4), carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 24-2 (CA 24-2), carbohydrate antigen 50 (CA 50), carcinoembryonic antigen (CEA), and gastrin 17 (G-17).
The kit for detecting gastric cancer is mainly a kit, and the kit comprises a reagent for detecting the content of SCTAG2 in a sample of a subject; the gastric cancer detection comprises gastric cancer screening, gastric cancer diagnosis, gastric cancer monitoring, gastric cancer medication guidance, gastric cancer prognosis judgment and/or the like. In some embodiments, the reagent is a nucleic acid level detection reagent or a protein level detection reagent. Further, when the reagent is at the nucleic acid level, the detection reagent is used to perform any one of the following methods: polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid sequencing, nucleic acid separation chip detection, denaturing high performance liquid chromatography, in situ hybridization, biological mass spectrometry, and HRM methods. Further, when the agent is at the protein level, the detection agent is used to perform any one of the following methods: biological mass spectrometry, amino acid sequencing, electrophoresis, and detection with antibodies specifically designed for the mutation site. In some embodiments, the kit further comprises reagents for detecting other components for use in a combination assay, including but not limited to: protamine antigen 1 (SCTAG 2), pepsinogen I/II, carbohydrate antigen 72-4 (CA 72-4), carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 24-2 (CA 24-2), carbohydrate antigen 50 (CA 50), carcinoembryonic antigen (CEA), and gastrin 17 (G-17). In some embodiments, the kit further comprises a sample processing reagent comprising at least one of a sample lysing reagent, a sample purifying reagent, and a sample nucleic acid or protein extracting reagent.
The following describes the technical content of the present invention in detail with reference to specific examples.
EXAMPLE 1 expression of SCTAG2 protein in stomach adenocarcinoma tissue
The study collects paraffin tissue sections of gastric cancer patients who were diagnosed at the daily friendship hospital in Jilin university between 6 months 2014 and 6 months 2017, comprising: 120 gastric adenocarcinoma histopathological sections and 96 normal gastric mucosa histopathological sections. Immunohistochemistry was used to detect the expression of SCTAG2 in the tissue. While their associated clinical pathology data including age, sex, degree of tumor cell differentiation, depth of infiltration (T), lymph node metastasis (N), clinical stage (TNM), general tumor typing, tumor size, vascular and neurological involvement, and expression of gastric adenocarcinoma tumor markers EGFR, p53 and HER-2 were counted.
The immunohistochemical results showed: SCTAG2 protein is widely expressed in stomach adenocarcinoma tissues, and the positive rate is 48.33% (58/120). The SCTAG2 protein is mainly expressed in cytoplasm of gastric adenocarcinoma cells, while the expression of the SCTAG2 protein is not found in normal gastric mucosa tissues, which indicates that the expression of the SCTAG2 in gastric adenocarcinoma tissues has higher specificity and is a reliable diagnostic marker (figure 1).
Furthermore, the study collates and counts the clinical pathological information of gastric cancer patients, and the correlation between the malignancy degree of the tumor and the expression positive rate of the SCTAG2 protein is analyzed by using a chi-square test. The results show that the positive rate of SCTAG2 protein expression in gastric cancer tissues is positively correlated with clinical stage (p=0.022) and tumor volume (p=0.046) of patients, namely, the later the clinical stage is, the higher the positive rate of SCTAG2 protein in gastric adenocarcinoma tissues with larger tumor volume is. Because patients with gastric adenocarcinoma with late clinical stage and large tumor volume are extremely easy to have local recurrence and distant metastasis, the treatment effect is poor and the prognosis is poor, the expression level of the SCTAG2 protein in gastric adenocarcinoma tissues can be used as a marker for evaluating the prognosis of gastric adenocarcinoma.
TABLE 1 relationship between the expression positive rate of SCTAG2 protein in gastric adenocarcinoma tissue and the clinical pathological characteristics
※ P is less than 0.05, has statistical significance
EXAMPLE 2 expression of SCTAG2 mRNA in stomach adenocarcinoma tissue
The study collected 20 pairs of freshly frozen gastric adenocarcinoma paired tissue samples, including cancer tissue, paracancerous tissue (2-5 cm from tumor margin) and normal tissue (more than 5cm from tumor margin), were subjected to real-time fluorescent quantitative PCR to detect the expression level of SCTAG2 in the tissue (fig. 2).
The results show that SCTAG2 expression levels were significantly elevated in cancer and paracancerous tissues, above normal tissues. The SCTAG2 has a certain correlation with the occurrence and development of gastric cancer, and the expression of the SCTAG2 in gastric cancer tissues has tissue specificity, so that the SCTAG2 can be a specific marker for diagnosing gastric cancer.
The foregoing descriptions of specific exemplary embodiments of the present invention are presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application to thereby enable one skilled in the art to make and utilize the invention in various exemplary embodiments and with various modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Sequence listing
<110> Chen Fangfang
<120> application of SCTAG2 in gastric cancer diagnosis and targeting drug
<160> 2
<170> SIPOSequenceListing 1.0
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<211> 55
<212> PRT
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
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Met Ser Thr Ser Arg Lys Leu Lys Ser His Gly Met Arg Arg Ser Lys
1 5 10 15
Ser Arg Ser Pro His Lys Gly Val Lys Arg Gly Gly Ser Lys Arg Lys
20 25 30
Tyr Arg Lys Gly Asn Leu Lys Ser Arg Lys Arg Gly Asp Asp Ala Asn
35 40 45
Arg Asn Tyr Arg Ser His Leu
50 55
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<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
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atgtcgacca gccgcaaatt aaagagtcat ggcatgagga ggagcaagag ccgatctcct 60
cacaagggag tcaagagagg tggcagcaaa agaaaatacc gtaagggcaa cctgaaaagt 120
aggaaacggg gcgatgacgc caatcgcaat taccgctccc acttgtga 168

Claims (10)

  1. Use of an SCTAG2 protein or a nucleic acid encoding an SCTAG2 protein as a diagnostic marker for the preparation of a diagnostic or diagnostic-aid detector for gastric cancer.
  2. 2. Use according to claim 1, characterized in that: the amino acid sequence of the SCTAG2 protein is shown as SEQ ID NO. 1; the sequence of the nucleic acid for encoding the SCTAG2 protein is shown as SEQ ID NO. 2.
  3. 3. Use according to claim 1, characterized in that: the detection agent is a reagent for detecting the expression of the SCTAG2 protein, and the reagent is an antibody for resisting the SCTAG2 protein; more preferably, the antibody against SCTAG2 protein is a monoclonal antibody; the detection agent is an expression agent for detecting a nucleic acid encoding an SCTAG2 protein, the agent being a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out the nucleic acid encoding the SCTAG2 protein; more preferably, the polynucleotide is an siRNA or shRNA, or a guide rna for a CRISPR/Cas9 system.
  4. 4. Use according to claim 2, characterized in that: the detection is quantitative detection or qualitative detection; the detection of SCTAG2 protein expression refers to detection of whether SCTAG2 protein expression is positive or negative. Detecting expression of a nucleic acid encoding an SCTAG2 protein refers to detecting whether expression of the nucleic acid encoding an SCTAG2 protein is positive or negative.
  5. 5. Use according to claim 2, characterized in that: when the expression of the SCTAG2 protein is detected as positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is detected as positive: prompting the risk (higher risk) or positive of gastric cancer; or to indicate a pre-cancerous pre-warning risk of gastric cancer, for example to indicate a risk of progression of a pre-cancerous lesion of gastric cancer (higher risk).
  6. 6. Use according to claim 1, characterized in that: the sample used by the detection agent is a tissue sample of a detected object; preferably, a surgical tissue sample or a biopsy sample such as an endoscopic biopsy sample; such stomach cancers include, but are not limited to, gastric adenocarcinoma, adenosquamous carcinoma, squamous carcinoma, and carcinoid.
  7. 7. Use according to claim 1, characterized in that: the use is the use for preparing a diagnostic reagent for gastric cancer, wherein the risk (higher risk) of gastric cancer or the positivity of gastric cancer is indicated when the expression of the SCTAG2 protein is positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is positive. Preferably, further screening such as histopathological examination is performed; the use is the use for preparing a detection agent for predicting the disease progression risk of gastric precancerous lesions, wherein the detection agent prompts the disease progression risk (higher risk) of gastric precancerous lesions when the expression of the SCTAG2 protein is positive and/or the expression of the nucleic acid encoding the SCTAG2 protein is positive; preferably, further screening such as histopathological examination is performed.
  8. 8. A test kit comprising one of the following test agents: (1) an agent that detects the expression of SCTAG2 protein; (2) An agent that detects expression of a nucleic acid encoding an SCTAG2 protein; the detection kit is used for screening gastric cancer, diagnosing gastric cancer, monitoring gastric cancer, guiding gastric cancer medication and/or prognosis of gastric cancer; the kit comprises reagents for detecting other early warning markers related to the risk of gastric precancerous lesions in addition to the reagents (1) and (2), preferably, other reagents include, but are not limited to, protamine antigen 1 (SCTAG 2), carbohydrate antigen 72-4 (CA 72-4), carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 24-2 (CA 24-2), carbohydrate antigen 50 (CA 50), carcinoembryonic antigen (CEA) and gastrin 17 (G-17); the kit further includes a sample processing reagent including at least one of a sample lysing reagent, a sample purifying reagent, and a sample nucleic acid or protein extracting reagent.
  9. 9. Use of any one selected from the following items (1) to (4) in the manufacture of a medicament for the treatment or co-treatment or prevention of gastric cancer: (1) SCTAG2 protein; (2) a nucleic acid encoding an SCTAG2 protein; (3) An agent that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein; (4) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein; the agent for reducing the expression level of SCTAG2 protein or inhibiting the expression of SCTAG2 protein in item (3) is an antibody against SCTAG2 protein; preferably, the antibody against SCTAG2 protein is a monoclonal antibody; the agent for reducing the expression level of a nucleic acid encoding an SCTAG2 protein or inhibiting the expression of a nucleic acid encoding an SCTAG2 protein as described in item (4) is a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out a nucleic acid encoding an SCTAG2 protein; preferably, the polynucleotide is an siRNA or shRNA, or a guide rna for a CRISPR/Cas9 system.
  10. 10. A medicament comprising the following item (1) or item (2), (1) a medicament that reduces the expression level of SCTAG2 protein or inhibits the expression of SCTAG2 protein; (2) An agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein; the agent that reduces the expression level of a nucleic acid encoding an SCTAG2 protein or inhibits the expression of a nucleic acid encoding an SCTAG2 protein is a nucleic acid construct comprising a polynucleotide for completely knocking out or partially knocking out a nucleic acid encoding an SCTAG2 protein; preferably, the polynucleotide is an siRNA or shRNA, or is a guide rna for a CRISPR/Cas9 system, the pharmaceutical composition further comprising one or more pharmaceutically acceptable excipients.
CN202210186194.3A 2022-02-28 2022-02-28 Application of SCTAG2 in gastric cancer diagnosis and targeting drugs Pending CN116699134A (en)

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