Application of urine secretion type Ly-6 / uPAR related protein (SCURP1) and immune globulin lambda variable 3-91 (IGLV3-9) in diagnosis and treatment of prostatic cancer
SLURP1 and IGLV3-9 in urine were screened through liquid phase secondary mass spectrometry, and detection kits were developed to solve the problems of early diagnosis and identification of prostate cancer in the prior art, and non-invasive and convenient urine marker detection was achieved, reducing examination costs and patient burden.
Patent Information
- Application Number
- CN202311497064.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-10
- Publication Date
- 2025-05-13
AI Technical Summary
It is difficult for the prior art to effectively use urine markers for early diagnosis and differential diagnosis of prostate cancer, and routine examination methods have problems such as high cost, strong invasiveness, and great psychological and economic burden on patients.
Protein spectrometry was established in urine samples of healthy people, prostate hyperplasia and prostate cancer patients through liquid phase secondary mass spectrometry (LC-MS/MS), and differentially expressed urine protein markers SLURP1 and IGLV3-9 were screened out, and corresponding detection kits were developed for the diagnosis, differentiation, diagnosis of condition, evaluation of treatment effect, monitoring and prognosis evaluation of prostate cancer.
It realizes efficient detection of SLURP1 and IGLV3-9 in urine, which can be used for the early diagnosis and identification of prostate cancer, reduces the cost of examination and the psychological and economic burden of patients, and provides a non-invasive and convenient diagnostic method.
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Abstract
Description
Technical Field
[0001] The present invention relates to the field of proteomics research, and specifically to the application of secretory Ly-6 / uPAR-related protein (SLURP1) and immunoglobulin lambda variable 3-91 (IGLV3-9) in urine in the preparation of reagents for prostate cancer diagnosis, differential diagnosis, disease severity assessment, treatment effect evaluation, monitoring, prognosis assessment and mechanism research. Background Art
[0002] Prostate cancer is a serious disease that endangers men's health worldwide. The clinical progression of prostate cancer is slow, and most patients have no obvious symptoms in the early stage. As prostate cancer develops, clinical symptoms such as dysuria, dysuria, and frequent urination are often manifested, which are similar to the symptoms of benign prostatic hyperplasia. At present, the screening and diagnosis of PCa are mainly based on blood PSA detection, rectal examination, imaging examination, and prostate puncture biopsy. However, benign prostatic hyperplasia, prostate inflammation or biopsy can lead to elevated PSA, and in terms of pathological tissue structure, the disease often leads to irregular prostate morphology, making it difficult to distinguish prostate cancer from the partition structure. In my country, most prostate cancer patients are already in the middle and late stages when they seek medical treatment, and the cost of the above-mentioned examinations and invasive punctures cause psychological and economic burdens on patients. Therefore, it is necessary to find relatively easy-to-obtain and patient-acceptable early diagnosis and differential diagnosis indicators for prostate cancer in clinical practice, especially for those with a family history of prostate cancer and elderly men. It has important practical significance.
[0003] Urine is the final metabolic product of blood. Changes in its composition, quantity and properties carry various information about the occurrence, development and prognosis of urinary system diseases. Compared with other body fluid samples such as serum, urine has the advantages of non-invasive acquisition and convenient collection. Prostate cancer cells and substances produced by their metabolism can enter urine from the prostate, which is of great significance for the diagnosis and identification of prostate cancer. However, there are currently few related biomarkers. Therefore, there is an urgent need to develop efficient urine markers for the diagnosis and identification of prostate cancer. Summary of the invention
[0004] In order to solve the above problems, the purpose of the present invention is to provide one or two urine protein markers SLURP1 and IGLV3-9 for use in the preparation of preparations for prostate cancer diagnosis, differential diagnosis, disease severity assessment, treatment effect evaluation, monitoring, prognosis assessment and mechanism research.
[0005] Preferably, the amino acid sequences of SLURP1 and IGLV3-9 in the urine are shown in SEQ ID NO.1 and SEQ ID NO.2 respectively. (SEQ ID NO.1: MASRWAVQLL LVAAWSVGCG EALKCYTCKE PMTSASCRTI TRCKPEDTACMTLVVTVEAE YPFNQSPVVT RSCSSSCVAT DPDSIGAAHL IFCCFRDLCN SEL) (SEQ ID NO.2: SYELTQPLSV SVALGQTARI TCGGNNIGSK NVHWYQQKPG QAPVLVIYRDSNRPSGIPER FSGSNSGNTA TLTISRAQAG DEADYYCQVW DSSTA) Preferably, the preparation is a kit for detecting the expression levels of SLURP1 and IGLV3-9 in the urine of prostate cancer patients.
[0006] Preferably, the preparation kit includes one or more of an immunological method of antigen-antibody reaction and a kit thereof, such as aptamer antibodies or antibody fragments that can specifically bind to SLURP1 and IGLV3-9.
[0007] Preferably, the kit further comprises components selected from the following group: any one or a combination of solid phase carrier, diluent, reference substance, standard substance, quality control substance, detection antibody, second antibody, second antibody diluent, luminescent reagent, washing solution, color developing solution and stop solution.
[0008] Preferably, the standards include SLURP1 and IGLV3-9 standards, and humanized tag antibody standards; preferably, the quality control products include: SLURP1 and IGLV3-9 quality control products, and humanized tag antibody quality control products; preferably, the solid phase carrier includes: microparticles, microspheres, glass slides, test strips, plastic beads, liquid chips, microplates or affinity membranes, etc. and other carriers with equivalent functions.
[0009] Preferably, the material of the solid phase carrier is any one of polyvinyl chloride, polystyrene, polyacrylamide, cellulose, and carriers with similar functions.
[0010] The inventors first collected urine samples from healthy people, patients with prostate hyperplasia, and prostate cancer, and established urine protein profiles for each group of subjects using liquid chromatography tandem mass spectrometry (LC-MS / MS). The data obtained from the mass spectrometry of the prostate cancer group, prostate hyperplasia group, and normal control group were quantitatively calculated, and finally the differential proteins SLURP1 and IGLV3-9 were obtained.
[0011] The present invention verifies through research that compared with healthy people, SLURP1 and IGLV3-9 in urine are highly expressed in urine of prostate cancer patients, which can be used for disease diagnosis; compared with prostate hyperplasia, SLURP1 and IGLV3-9 are also highly expressed in urine of prostate cancer patients, which can be used for disease identification. Therefore, it is proposed that the detection of SLURP1 and IGLV3-9 in urine can be used for the diagnosis, differential diagnosis, judgment of disease severity, evaluation of treatment effect, monitoring, prognosis evaluation and mechanism research of prostate cancer.
[0012] The present invention takes advantage of the non-invasive acquisition of urine specimens, large-scale repeated sampling, and convenient storage, and uses urine specimens to detect SLURP1 and IGLV3-9 in urine.
[0013] In order to make the above and other purposes, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are given below and described in detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] Figure 1 This is a graph showing the levels of SLURP1 and IGLV3-9 in urine in prostate cancer, prostate hyperplasia and healthy control groups.
[0015] Figure 2 Western blotting and ELISA were used to detect the expression of SLURP1 in urine in prostate cancer group, prostate hyperplasia and healthy control group.
[0016] Figure 3 It is an evaluation of the diagnostic and differential efficacy of SLURP1 in urine for prostate cancer. Implementation
[0017] Example 1 Urine specimen collection and processing Patients with prostate cancer and prostate hyperplasia were selected as the prostate cancer group and prostate hyperplasia group, and healthy subjects who underwent physical examination during the same period were selected as the normal control group. Fresh morning urine samples were collected from the subjects in each group after admission, and urine from the morning catheter was collected from those who could not urinate normally and placed in a dry and clean container.
[0018] The urine samples were subjected to protein extraction and enzymatic desalination, and the extracted protein concentration was measured. The urine protein profiles of different groups were constructed using advanced liquid chromatography-mass spectrometry (LC-MS / MS).
[0019] Example 2 Differential peptide data analysis All experiments were performed three times, and all values are expressed as mean ± standard deviation. The analysis results of differentially expressed proteins were obtained by fold change 1.2. PA value <0.05 was considered statistically significant, and differentially expressed proteins were screened, and global cluster analysis was performed on the target proteins.
[0020] Compared with healthy people, SLURP1 and IGLV3-9 in urine are highly expressed in the urine of prostate cancer patients, which can be used for disease diagnosis; compared with prostate hyperplasia, SLURP1 and IGLV3-9 are also highly expressed in the urine of prostate cancer patients, which can be used for disease identification. Figure 1 As shown, there is a significant difference in expression.
[0021] Example 3 Validation of SLURP1 in urine Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to detect SLURP1 in the urine of healthy people and prostate cancer patients. Figure 2 As shown. Compared with healthy people, SLURP1 is highly expressed in the urine of prostate cancer patients; compared with prostate hyperplasia, SLURP1 is also highly expressed in the urine of prostate cancer patients. ( p <0.01). ROC curve analysis showed that SLURP1 had high diagnostic and identification sensitivity and specificity for prostate cancer.
[0022] Although the present invention has been disclosed as above in terms of preferred embodiments, it is not intended to limit the present invention. Any technician in the relevant technical field may make some changes and improvements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be based on what is defined in the claims.
Claims
1. The use of urine protein markers in the preparation of preparations for prostate cancer diagnosis, differential diagnosis, disease severity assessment, treatment effect evaluation, monitoring, prognosis assessment and mechanism research, characterized in that: The biomarkers include one or two of the following proteins: secretory urinary Ly-6 / uPAR-related protein (SLURP1) and immunoglobulin lambda variable 3-91 (IGLV3-9).
2. The use according to claim 1, characterized in that: The biomarkers include one or two thereof; preferably, the biomarkers include at least SLURP1, and further preferably, the biomarkers include SLURP1 and IGLV3-9.
3. The use according to claim 1, characterized in that: The amino acid sequences of the urine SLURP1 and IGLV3-9 are shown in SEQ ID NO: 1-2.
4. The use according to claim 1, characterized in that: The preparation is used to detect the expression level of either or both of SLURP1 and IGLV3-9.
5. The use according to claim 4, characterized in that: in, The preparation is used for prostate cancer diagnosis, differential diagnosis, disease severity assessment, treatment effect evaluation, monitoring, prognosis assessment and mechanism research. And / or, the preparation includes antigen-antibody detection immunoreagent; preferably, it includes urine antigen-antibody detection immunoreagent.
6. The use according to claim 5, characterized in that: The reagent preparation kit includes an immunological method for antigen-antibody reaction and a kit thereof, and can specifically bind to one or more of SLURP1 and IGLV3-9 aptamer antibodies or antibody fragments.
7. The use according to claim 5, characterized in that: The kit also includes components selected from the following: any one or a combination of solid phase carrier, diluent, reference substance, standard substance, quality control substance, detection antibody, second antibody, second antibody diluent, luminescent reagent, washing solution, color developing solution, and stop solution.
8. The use according to claim 9, characterized in that: The standard products include SLURP1 and IGLV3-9 standard products, and humanized tag antibody standard products; the quality control products include: SLURP1 and IGLV3-9 quality control products, and humanized tag antibody quality control products; the solid phase carrier includes: microparticles, microspheres, glass slides, test strips, plastic beads, liquid chips, microplates or affinity membranes, as well as other carriers with equivalent functions.
9. The use according to claim 9, characterized in that: The material of the solid phase carrier is any one of polyvinyl chloride, polystyrene, polyacrylamide, cellulose and carriers with similar functions.