CN110734979B - Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient - Google Patents
Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient Download PDFInfo
- Publication number
- CN110734979B CN110734979B CN201911111701.1A CN201911111701A CN110734979B CN 110734979 B CN110734979 B CN 110734979B CN 201911111701 A CN201911111701 A CN 201911111701A CN 110734979 B CN110734979 B CN 110734979B
- Authority
- CN
- China
- Prior art keywords
- stamp
- gene
- mrna
- multiple myeloma
- patients
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Oncology (AREA)
- Biochemistry (AREA)
- Hospice & Palliative Care (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses application of OC-STAMP as a marker for evaluating the prognosis risk of multiple myeloma patients. According to the invention, the expression level of OC-STAMP in bone marrow cells of an MM patient and a healthy donor is analyzed through an RT-qPCR method, and the expression level of the gene in the MM patient is found to be remarkably higher than that of a normal control and possibly related to clinical course. Subsequent studies of the clinical significance of this gene revealed that high expression of this gene was an independent risk factor for low Progression Free Survival (PFS) in MM patients. OC-STAMPs are of great value in assessing prognostic risk in patients with multiple myeloma.
Description
Technical Field
The invention belongs to the technical field of medical detection, and particularly relates to application of OC-STAMP as a marker for evaluating prognosis risk of multiple myeloma patients.
Background
Multiple Myeloma (MM) is the second most common hematological malignancy that originates from plasma cells, accounting for 10-15% of all hematological tumors. Myeloma cells proliferate maliciously in the bone marrow and secrete monoclonal immunoglobulins, which can cause organ and tissue damage. Infiltration of bone marrow can lead to suppression of normal hematopoiesis and the appearance of anemia, infection, and bleeding. Infiltration in bone can stimulate osteoclasts to enhance their osteolytic action, which can lead to bone pain, bone defects, hypercalcemia, and pathological fractures. The disease is well developed in the elderly, the average age of the disease is about 70 years old, and the proportion of male and female is (1.6-3): 1. The current treatment of MM is predominantly drug therapy. In terms of prognosis stratification and curative effect judgment, cytogenetic indexes and biochemical indexes are often relied on, and specific molecular biological markers are lacked.
OC-STAMP (osteo plastic proteins-membrane protein) is a novel gene found in osteoclasts, whose expression is up-regulated during differentiation of monocytes into osteoclasts. The gene expression product of OC-STAMP has high similarity with DC-STAMP (cell Expressed derived Transmembrane protein) protein family, is conserved at the carboxyl terminal, and has a main function as a key molecule in the process of fusing and differentiating osteoclasts. Recent studies have shown that the OC-STAMP gene encodes a cell membrane anchoring receptor under induction of RANKL (receptor activator of nuclear factor-. kappa.b ligand) and interacts with DC-STAMP during monocyte fusion to form osteoclasts.
Disclosure of Invention
It is an object of the present invention to assess the prognostic risk of multiple myeloma patients.
The invention firstly protects the new application of a substance for detecting OC-STAMP genes or a substance for detecting mRNA encoded by the OC-STAMP genes or a substance for detecting protein encoded by the OC-STAMP genes.
The invention provides an application of a substance for detecting OC-STAMP gene or a substance for detecting mRNA encoded by the OC-STAMP gene or a substance for detecting protein encoded by the OC-STAMP gene in any one of the following (1) to (6):
(1) preparing a product for prognostic risk assessment of multiple myeloma patients;
(2) assessing a prognostic risk for multiple myeloma patients;
(3) preparing a product for prognosis of progression-free survival assessment of multiple myeloma patients;
(4) assessing the prognosis for progression-free survival in patients with multiple myeloma;
(5) preparing a product for the prognostic assessment of progression-free survival time in patients with multiple myeloma;
(6) assessing progression-free survival time for multiple myeloma patients.
The invention also protects products for the prognostic risk and/or progression-free survival assessment of multiple myeloma patients.
The product for evaluating the prognosis risk and/or progression-free survival rate and/or progression-free survival time of multiple myeloma patients comprises a substance for detecting OC-STAMP gene or a substance for detecting mRNA encoded by the OC-STAMP gene or a substance for detecting protein encoded by the OC-STAMP gene.
The product for the prognosis of risk and/or progression-free survival assessment of multiple myeloma patients further comprises a data processing device; the data processing device is internally provided with a module; the module has the functions as shown in (a1) and (a 2):
(a1) taking in-vitro bone marrow of a to-be-detected population consisting of a plurality of multiple myeloma patients without any treatment measures as a sample, determining the expression quantity of mRNA (messenger ribonucleic acid) coded by an OC-STAMP gene in each sample, arranging and halving the to-be-detected population according to the sequence of the expression quantity of the mRNA from low to high, taking 1/2 with low expression quantity as an OC-STAMP gene mRNA low expression group, and taking the rest 1/2 of the to-be-detected population as an OC-STAMP gene mRNA high expression group;
(a2) determining the prognostic risk and/or the prognostic progression-free survival rate and/or the progression-free survival time of a test subject from said test population according to the following criteria: the prognosis risk of the testee in the OC-STAMP gene mRNA low-expression group is lower or the candidate is lower than that of the testee in the OC-STAMP gene mRNA high-expression group; the prognosis non-progression survival rate of the testee in the OC-STAMP gene mRNA low expression group is higher than or candidate higher than that of the testee in the OC-STAMP gene mRNA high expression group; the prognosis progression-free survival time of the testees in the OC-STAMP gene mRNA low expression group is longer than or candidate to be longer than that of the testees in the OC-STAMP gene mRNA high expression group.
In any of the above applications or products, the material for detecting the expression level of the OC-STAMP gene can be a reagent and/or an apparatus required for detecting the expression level of the OC-STAMP gene by using a method in the prior art, such as a reagent and/or an apparatus required for detecting the expression level of the OC-STAMP gene by using high-throughput sequencing, or a reagent and/or an apparatus required for detecting the expression level of the OC-STAMP gene by using quantitative PCR.
Furthermore, the substance for detecting the OC-STAMP gene is a substance for detecting mRNA encoded by the OC-STAMP gene. The substance for detecting mRNA encoded by the OC-STAMP gene can comprise specific amplification primers and/or probes for detecting the mRNA of the OC-STAMP gene. In a specific embodiment of the invention, the specific amplification primers for detecting the mRNA of the OC-STAMP gene are primer pairs consisting of two single-stranded DNAs shown as a sequence 1 and a sequence 2 in a sequence table; the probe for detecting the mRNA of the OC-STAMP gene is a single-chain probe shown as a sequence 3 in a sequence table, the 5 'end of the single-chain probe can be marked with a fluorescence reporter group (such as FAM), and the 3' end of the single-chain probe can be marked with a fluorescence quenching group (such as BHQ).
Furthermore, the substance for detecting mRNA encoded by the OC-STAMP gene can also comprise specific amplification primers and/or probes for detecting mRNA of an internal reference gene. The reference gene may specifically be the ABL1 gene. In one specific embodiment of the invention, the specific amplification primer for detecting the mRNA of the ABL1 gene is a primer pair consisting of two single-stranded DNAs shown as a sequence 4 and a sequence 5 in a sequence table; the probe for detecting the mRNA of the ABL1 gene is a single-chain probe shown as a sequence 6 in a sequence table, the 5 'end of the single-chain probe can be marked with a fluorescence reporter group (such as FAM), and the 3' end of the single-chain probe can be marked with a fluorescence quenching group (such as BHQ).
The invention also discloses a method for evaluating the prognosis risk of a multiple myeloma patient.
The method for evaluating the prognosis risk of the multiple myeloma patient comprises the following steps: detecting the expression level of the OC-STAMP gene in the mononuclear cells of a patient to be detected: the prognosis risk of the test patient with low expression level of the OC-STAMP gene in the mononuclear cell is lower than or is lower than that of the test patient with high expression level of the OC-STAMP gene in the mononuclear cell.
The invention finally provides a method for evaluating the prognosis progression-free survival rate and/or progression-free survival time of a multiple myeloma patient.
The method for evaluating the prognosis progression-free survival rate and/or progression-free survival time of the multiple myeloma patients comprises the following steps: detecting the expression level of the OC-STAMP gene in the mononuclear cells of a patient to be detected: the prognosis non-progression survival rate of the patient to be tested with low expression level of the OC-STAMP gene in the mononuclear cell is higher than or is candidate to be higher than that of the patient to be tested with high expression level of the OC-STAMP gene in the mononuclear cell; the test patient having a low expression level of the OC-STAMP gene in the mononuclear cell has a longer prognosis progression-free survival time than or is candidate for a test patient having a higher expression level of the OC-STAMP gene in the mononuclear cell.
In any of the above methods, the detecting the expression level of the OC-STAMP gene in the mononuclear cells of the patient to be tested can be performed by using the substance for detecting the mRNA encoded by the OC-STAMP gene.
The application of the OC-STAMP gene as a target in the preparation of the kit also belongs to the protection scope of the invention; the application of the kit is any one of the following (1) to (6):
(1) preparing a product for prognostic risk assessment of multiple myeloma patients;
(2) assessing a prognostic risk for a patient with multiple myeloma;
(3) preparing a product for prognosis of progression-free survival assessment of multiple myeloma patients;
(4) assessing progression-free survival prognosis for multiple myeloma patients;
(5) preparing a product for the prognostic assessment of progression-free survival time in patients with multiple myeloma;
(6) assessing progression-free survival time for multiple myeloma patients.
In the present invention, the prognostic risk is the prognostic risk of multiple myeloma patients at initial diagnosis. The high or low of the prognostic risk can be embodied as all or part of the following indexes: progression free survival time, progression free survival rate. The longer the progression-free survival, the lower the prognostic risk; the higher the progression-free survival, the lower the risk of prognosis.
In the present invention, the OC-STAMP gene Sequence is shown as Sequence 9 in the Sequence Listing (NCBI Reference Sequence: NM-080721.3).
According to the invention, the OC-STAMP expression level in bone marrow cells of an MM patient is analyzed through an RT-qPCR method, and the high expression of the OC-STAMP is found to be an independent risk factor of low-progression-free survival (PFS) of the MM patient for the first time by combining clinical course and survival data. OC-STAMPs are of great value in assessing prognostic risk in patients with multiple myeloma.
Drawings
FIG. 1 shows the expression levels of OC-STAMP in bone marrow cells of naive and remitted MM patients and healthy donors. Lines represent median and quartile values; p < 0.0001.
FIG. 2 is a ROC plot of the OC-STAMP diagnostic first-visit MM.
FIG. 3 is a graph of the relationship between OC-STAMP expression level and clinical course of MM. Expression levels of OC-STAMP in bone marrow mononuclear cell specimens for initial diagnosis, remission and disease progression in 5 MM patients.
FIG. 4 is a graph of the relationship of OC-STAMP expression level to MM prognostic progression-free survival.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
The OC-STAMP gene Sequence in the invention is shown as Sequence 9 in the Sequence table (NCBI Reference Sequence: NM-080721.3).
Example 1 use of OC-STAMP as a marker for assessing the prognostic risk of multiple myeloma patients
Study object and method
1. Study object
Bone marrow specimens of 160 cases of multiple myeloma patients (all patients informed consent) collected by the institute of hematological diseases in the national hospital of Beijing university were collected as study subjects from 1 month to 2016 (1 month to 2016) in 2014, including 100 cases of men, 60 cases of women, the median age of 60 years, and the age range of 33 to 87 years, and were followed up to death, missed visits, or 8 months in 2019. The diagnostic criteria for multiple myeloma are referenced to National Comprehensive Cancer Network (NCCN) guidelines, and staging is performed according to the Durie-Salmonon (D-S) staging system, the International Staging System (ISS), and the revised International staging System (R-ISS). Treatment and efficacy assessment of multiple myeloma patients was performed with reference to the above guidelines. Progression Free Survival (PFS) is defined as the time from the start of the initial treatment to the first occurrence of disease progression, which is an event.
In addition, 60 bone marrow specimens were collected during follow-up of 55 naive multiple myeloma patients, including 55 specimens with Complete Remission (CR), Very Good Partial Remission (VGPR), Partial Remission (PR), and 5 specimens with disease progression after treatment. Normal control bone marrow specimens were obtained from adult healthy volunteers in a total of 42 cases. The specimens used in the study protocol were approved by the ethical committee of the people's hospital of Beijing university. All healthy volunteers and patients signed informed consent.
2. Extraction of bone marrow mononuclear cells and RT-qPCR
Mononuclear cells in a bone marrow specimen are separated by Ficoll lymphocyte separation and density gradient centrifugation, RNA is extracted from the mononuclear cells and is reversely transcribed into cDNA. And (2) performing RT-qPCR by using cDNA as a template and adopting an OC-STAMP primer pair (the OC-STAMP primer pair consists of an OC-STAMP upstream primer and an OC-STAMP downstream primer, the size of an amplification product is 101bp, and the nucleotide sequence is shown as a sequence 7) and an OC-STAMP probe.RT-qPCR was performed using cDNA as a template and ABL1 primer pair (ABL1 primer pair consisting of ABL1 upstream primer and ABL1 downstream primer, amplification product size 124bp, nucleotide sequence shown in sequence 8) and ABL1 probe. A10. mu.L PCR reaction system was configured using the PCR Master Mix kit as follows: 5 μ L
Universal PCR Master Mix; upstream primer 0.9. mu.M, downstream primer 0.9. mu.M, probe 0.25. mu.M; 150-500ng cDNA, primer sequence and fluorescent probe sequence are shown in Table 1. qPCR was performed using ABI 7500FAST PCR amplification apparatus under the following reaction conditions: 50 ℃ for 2min, 95 ℃ for 10min, then 95 ℃ for 15s, 60 ℃ for 1min for 40 cycles. The copy numbers of OC-STAMP and ABL1 are calculated by a standard curve method by taking ABL1 as an internal reference. Serially diluted ABL 1-expressing plasmids (see "ABL 1 plasmid" in "Gabert J, Beillard E, van der Velden VH, Bi W, Grimwide D, Pallisgard N, et al. Standard and quality control of" real-time "quantitative reverse transcription polymerase reaction of fusion gene transformations for reactive separation in free-space-a Europe agricultural Cancer promoter. Leukemia.2003; 17: 2318- 6 、10 5 、10 4 、10 3 、10 2 、10 1 And 10 0 Copy/. mu.L) and OC-STAMP positive bone marrow samples were used to construct standard quantification curves. The curve threshold is set to 0.082. Ct values of ABL1 and OC-STAMP are obtained through amplification curves of reference genes ABL1 and OC-STAMP in a sample and a set threshold (0.082), copy numbers of the sample ABL1 and the sample OC-STAMP are obtained according to an ABL1 standard curve (because the amplification efficiency of the OC-STAMP is close to that of ABL1, calculation is carried out by referring to the ABL1 standard curve for reducing experimental errors), the copy number of the sample OC-STAMP is divided by the copy number of ABL1 to be the sample OC-STAMP expression quantity, and in order to be consistent with a form reported by clinical routine, the result is multiplied by one hundred percent, and finally the sample OC-STAMP expression quantity is presented in a form of the copy number of the OC-STAMP/copy number of the ABL1 multiplied by one percent.
TABLE 1 OC-STAMP and ABL1 primer sequences
3. Statistical analysis
Statistical analysis was performed using SPSS22.0, Graphpad Prism 7. And (3) comparing the differences of the two groups of data, wherein chi-square test is adopted for classified variable data, t test is adopted for continuous variable data, and the difference is less than 0.05, so that the statistical significance is achieved. Receiver Operating Characterization (ROC) curves were used to evaluate specificity and sensitivity of diagnostic markers. The Youden Index (Youden Index) is used to calculate the cut-off value for a diagnosis. Survival analysis was performed using Kaplan-Meier with log-rank test applied. The Cox proportional hazards regression model performs a multifactorial analysis. The variables with P more than 0.1 are gradually eliminated by a back-off method, and P less than 0.05 has statistical significance.
Second, research results
1. Expression levels of OC-STAMP in multiple myeloma
The expression level of OC-STAMP in bone marrow cells of a patient with preliminary-diagnosis multiple myeloma (median 0.52%; range 0-121%) is significantly higher than that of remission patients (0.01%; 0-0.4.7%; P < 0.0001) and healthy donor controls (0.02%; 0-0.10%; P < 0.0001%). While there was no significant difference in expression levels between remission patients and healthy donors (P ═ 0.85) (fig. 1).
ROC curve analysis showed that the area under the curve (AUC) of OC-STAMP diagnostic MM was 0.867 (95% confidence interval 0.820-0.915; P < 0.0001; FIG. 2), and the OC-STAMP expression level corresponding to the maximum approximate ascending index was 0.11%. When this is the diagnostic threshold of MM, the overexpression rate of OC-STAMP in primary MM is 69.375%.
2. OC-STAMP expression level is related to clinical course of multiple myeloma
In order to further study the relationship between OC-STAMP and the clinical course of adult multiple myeloma, 5 bone marrow specimens of adult multiple myeloma patients at the time of initial diagnosis, remission and disease progression were analyzed for the expression level of OC-STAMP.
The results showed that the level of OC-STAMP expression in bone marrow cells was significantly lower at full remission than at initial diagnosis, and significantly higher at relapse (disease progression) (fig. 3).
3. Relationship between OC-STAMP expression level and general clinical characteristics of multiple myeloma
160 patients with initial diagnosis of multiple myeloma were classified into OC-STAMP high-expression group and OC-STAMP low-expression group according to the median value of OC-STAMP expression level.
The results show that patients in the OC-STAMP high expression group are more likely to appear in the stages IIIA and IIIB (P is 0.035) in the D-S stage at the initial diagnosis, and the patients in the OC-STAMP high expression group are more likely to appear low serum albumin (less than 35g/L, P is 0.036), the bone marrow plasma cell proportion is more than or equal to 50% (P is 0.039), and more than 3 bone destruction (P is 0.000) is more likely to occur. In addition, patients in the OC-STAMP high expression group were more likely to develop high-risk cytogenetic markers (P0.014) associated with poor prognosis (table 2).
TABLE 2 relationship between OC-STAMP expression level and the general clinical features of preliminary-diagnosis adult multiple myeloma
4. Relationship between OC-STAMP expression level and prognosis of multiple myeloma patients
Survival analysis of 132 patients with prognostic information by Kaplan-Meier method revealed that 48-month progression-free survival rates (PFS) were 4.8% and 27.7% in OC-STAMP high-expression and low-expression patients, respectively, and that OC-STAMP high-expression patients had shorter progression-free survival times than low-expression patients (median 13.38vs.21.57 months; P ═ 0.0025; fig. 4). To further clarify the effect of OC-STAMP on prognosis, the following factors were included in the multifactorial analysis, due to the number of factors affecting MM prognosis: beta 2 microglobulin level (more than or equal to vs. < 3.5mg/L) in initial diagnosis, lactate dehydrogenase level (more than or equal to vs. < 245g/L) in initial diagnosis, and receiving hematopoietic stem cell transplantation (if v.no). The results show that high expression of OC-STAMP is an independent risk factor for progression-free survival (table 3).
TABLE 3 Multi-factor analysis of Progression Free Survival (PFS) in adult multiple myeloma patients
| End of business | Risk ratio (95% confidence interval) | P value |
| OC-STAMP: high expression vs. Low expression | 1.865(1.224-2.843) | 0.004* |
| |
1.257(0.779-2.026) | 0.348 |
| Lactate dehydrogenase levels:<245g/L vs.≥245g/L | 1.114(0.648-1.916) | 0.695 |
| receiving hematopoietic stem cell transplantation: is vs. no | 0.825(0.487-1.399) | 0.476 |
Sequence listing
<110> Beijing university Hospital (second clinical medical college of Beijing university)
Application of <120> OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient
<160>9
<170>PatentIn version 3.5
<210>1
<211>21
<212>DNA
<213>Artificial Sequence
<400>1
tgtggactgg gctcagaagt t 21
<210>2
<211>22
<212>DNA
<213>Artificial Sequence
<400>2
ttcatccctt tcctcttcaa cc 22
<210>3
<211>31
<212>DNA
<213>Artificial Sequence
<400>3
cacggtcaag tatgatgtgg catacactgt c 31
<210>4
<211>31
<212>DNA
<213>Artificial Sequence
<400>4
tggagataac actctaagca taactaaagg t 31
<210>5
<211>21
<212>DNA
<213>Artificial Sequence
<400>5
gatgtagttg cttgggaccc a 21
<210>6
<211>28
<212>DNA
<213>Artificial Sequence
<400>6
ccatttttgg tttgggcttc acaccatt 28
<210>7
<211>101
<212>DNA
<213>Artificial Sequence
<400>7
tgtggactgg gctcagaagt tgccaactgt gcccatcacg ctcacggtca agtatgatgt 60
ggcatacact gtcctgggct tcatcccttt cctcttcaac c 101
<210>8
<211>124
<212>DNA
<213>Artificial Sequence
<400>8
tggagataac actctaagca taactaaagg tgaaaagctc cgggtcttag gctataatca 60
caatggggaa tggtgtgaag cccaaaccaa aaatggccaa ggctgggtcc caagcaacta 120
catc 124
<210>9
<211>2123
<212>DNA
<213>Artificial Sequence
<400>9
agggattccc aagtcccagc gattccccca cttcctctcc acctgccaca gctgccagcg 60
accgcccgcc tgaaaccact gccatttgga cagcatgcca ggccacccag gagcagctga 120
gcaacttgtc aagaccgggt ggaggtcctg gcacttgggg ttctggaagg cccttgcccc 180
actgcaggct gcctgggacg ccttctccca gcctgttcca gccagctgtg gccagctgct 240
gacccagctc ctcctgtgtg cctccctggc tgctgctgct gcaggtctgg tttatcactg 300
gctggcatcc ttgctgcttt atcctcctgg accttcagcc atggttgcca ctgtctgtgg 360
cctcctggtc ttcctgagcc tgggcctggt acccccagtc cgctgcctgt ttgcactcag 420
cgtgcccacc ctgggtatgg agcagggccg ccggctgctc ctgtcctaca gcactgccac 480
cctggccatt gctgtggtgc ccaacgtcct ggccaacgtg ggtgcggccg ggcaggtgct 540
gaggtgtgtc accgagggct ccctggagag tctcctcaat accactcacc agctgcatgc 600
agcatccagg gctctgggcc ccacaggcca ggcaggcagc cggggcctga catttgaggc 660
ccaggacaat ggctctgcct tctaccttca catgctcagg gtcactcagc aggtcctgga 720
ggatttctct ggcctggagt ccctggcccg ggcagcagcg ctagggaccc agcgagtggt 780
cacagggctg tttatgttgg gcctcctggt ggagtcggca tggtacctcc attgctacct 840
gacagacctg cggtttgaca atatctacgc cactcaacag ctgacccagc ggttggcaca 900
ggcccaggct acacacctcc tggcccctcc acccacctgg ctgctccagg cggctcagct 960
gaggctgtca caggaggagc tgttgagttg tcttctaagg ctggggctgc ttgccctgct 1020
cctcgtggcc acggctgtgg cggtggccac agaccatgta gccttcctcc tggcacaggc 1080
tactgtggac tgggctcaga agttgccaac tgtgcccatc acgctcacgg tcaagtatga 1140
tgtggcatac actgtcctgg gcttcatccc tttcctcttc aaccagctgg ctccggagag 1200
ccccttcctc tccgtccaca gctcctacca atgggagctc cgcctcacct ccgcccgctg 1260
cccactgcta cccgcccggc gtccccgcgc agctgccccg ctggccgcgg gggccctgca 1320
gctcctggcg ggctccacgg tgctcctgga ggcctacgcc cgccgcctgc ggcatgccat 1380
cgccgcttcc ttcttcacag cccaggaggc gaggagggtc cgccacctgc acgcccggct 1440
ccagcgaaga cacgacaggc accaaggcca gcagctgccc ctaggggatc cttcttgcgt 1500
ccccacaccc agacctgcct gcaagcctcc ggcatggata gactacaggc tggatgcctt 1560
aagaaccgag agcagtgagg gagaagggaa agagctttgg agttgcagag acctgagttg 1620
taaccttggt cctgtgccgc ctccctgtgt gaccttgggt aagtcacttc acctctctga 1680
gcctcggttt ctacatctgc ataacgacag catatttacc attgatgtga cctacttccc 1740
acgcagggat gtggtcagga tggaaggaaa tactgggcat gataggcctg gataaccggt 1800
aaagaaccat gcaaaggcga agacaaggag tgcagagaga gctcatggtt cctccaggct 1860
ggttggcgat caggctcatc tcatctgcac caactgctct acttgttaga tggagacctt 1920
gcatcatgaa tttctcgaaa tgctcctgga acttatttat atgcctcaaa atcctctaaa 1980
ctcatttata gtaacccata gttttaattt tataaataaa cgtatttatt aaatcttaga 2040
ttacgttatt cctctgcctc aaattccttc agggcctttc cattgccagg attagatttt 2100
gccaaattaa aatataggac act 2123
Claims (9)
1. Detection ofOC-STAMPSubstance or detection of genesOC-STAMPAgents or assays for mRNA encoded by genesOC-STAMPUse of a substance of a gene-encoded protein in any one of the following (1) to (3):
(1) preparing a product for prognostic risk assessment of multiple myeloma patients;
(2) preparing a product for assessing the prognosis of progression-free survival rate of multiple myeloma patients;
(3) and preparing a product for evaluating the prognosis of progression-free survival time of the multiple myeloma patients.
2. Use according to claim 1, characterized in that: the detection is carried outOC-STAMPThe substance of mRNA encoded by the gene is detectedOC-STAMPPrimers and/or probes specific for the mRNA encoded by the gene.
3. Use according to claim 2, characterized in that: the specific amplification primer is a primer pair consisting of two single-stranded DNAs shown as a sequence 1 and a sequence 2 in a sequence table; the probe is a single-stranded probe shown as a sequence 3 in a sequence table.
4. Product for prognostic risk assessment in patients with multiple myeloma comprising detectionOC-STAMPSubstance or detection of genesOC-STAMPAgents or assays for mRNA encoded by genesOC-STAMPMaterials and data processing devices for gene-encoded proteins; a module is arranged in the data processing device; the module has the functions as shown in (a1) and (a 2):
(a1) taking ex vivo bone marrow of a to-be-detected population consisting of a plurality of patients with multiple myeloma who do not take any treatment measures as a sample, and determining the content of each sampleOC-STAMPThe expression quantity of mRNA coded by the gene is arranged and halved according to the sequence of the expression quantity of the mRNA from low to high, and 1/2 with low expression quantity is used as the population to be detectedOC-STAMPGene mRNA low expression group, using the rest 1/2 said testing population asOC-STAMPGene mRNA high expression group;
(a2) determining the risk of prognosis of a subject from said test population according to the following criteria: the above-mentionedOC-STAMPThe prognosis risk of the subject in the gene mRNA low expression group is lower than or lower than that of the candidateOC-STAMPSubjects in the gene mRNA high expression group.
5. A product for the prognostic assessment of progression-free survival in patients with multiple myeloma comprising a testOC-STAMPSubstance or detection of genesOC-STAMPAgents or assays for mRNA encoded by genesOC-STAMPMaterials and data processing devices for gene-encoded proteins; a module is arranged in the data processing device; the module has functions as shown in (b 1) and (b 2):
(b1) taking ex vivo bone marrow of a population to be tested consisting of a plurality of patients with multiple myeloma who do not take any treatment measures as a specimen, and determining the amount of each specimenOC-STAMPThe expression quantity of mRNA encoded by the gene is arranged according to the sequence of the expression quantity of the mRNA from low to high, and the population to be detected is arrangedBisection, using 1/2 to be tested with low expression as the populationOC-STAMPGene mRNA low expression group, using the rest 1/2 said testing population asOC-STAMPGene mRNA high expression group;
(b2) determining the prognostic progression-free survival of a test subject from said test population according to the following criteria: the above-mentionedOC- STAMPThe survival rate of the prognosis progression-free of the testee in the gene mRNA low expression group is higher or the candidate is higher than thatOC-STAMPSubjects in the gene mRNA high expression group.
6. A product for the prognostic assessment of progression-free survival time in patients with multiple myeloma comprising a testOC-STAMPSubstance or detection of genesOC-STAMPAgents or assays for mRNA encoded by genesOC-STAMPMaterials and data processing devices for gene-encoded proteins; a module is arranged in the data processing device; the module has functions as shown in (c 1) and (c 2):
(c1) taking ex vivo bone marrow of a to-be-detected population consisting of a plurality of patients with multiple myeloma who do not take any treatment measures as a sample, and determining the content of each sampleOC-STAMPThe expression quantity of mRNA coded by the gene is arranged and halved according to the sequence of the expression quantity of the mRNA from low to high, and 1/2 with low expression quantity is used as the population to be detectedOC-STAMPGene mRNA low expression group, using the rest 1/2 said population to be tested asOC-STAMPGene mRNA high expression group;
(c2) determining the progression free survival time of a test subject from said test population according to the following criteria: the above-mentionedOC- STAMPThe prognosis progression-free survival time of the subject in the gene mRNA low expression group is longer than or candidate to be longer than thatOC-STAMPSubjects in the gene mRNA high expression group.
7. The product according to any one of claims 4-6, wherein: the detection is carried outOC-STAMPThe substance of mRNA encoded by the gene is detectedOC-STAMPPrimers and/or probes specific for the mRNA encoded by the gene.
8. The product of claim 7, wherein: the specific amplification primer is a primer pair consisting of two single-stranded DNAs shown as a sequence 1 and a sequence 2 in a sequence table; the probe is a single-stranded probe shown as a sequence 3 in a sequence table.
9.OC-STAMPThe gene is used as a target to be applied to the preparation of the kit; the application of the kit is any one of the following (1) to (6):
(1) preparing a product for prognostic risk assessment of multiple myeloma patients;
(2) assessing a prognostic risk for a patient with multiple myeloma;
(3) preparing a product for assessing the prognosis of progression-free survival rate of multiple myeloma patients;
(4) assessing the prognosis for progression-free survival in patients with multiple myeloma;
(5) preparing a product for the prognostic assessment of progression-free survival time in patients with multiple myeloma;
(6) assessing progression-free survival time for multiple myeloma patients.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911111701.1A CN110734979B (en) | 2019-11-14 | 2019-11-14 | Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911111701.1A CN110734979B (en) | 2019-11-14 | 2019-11-14 | Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110734979A CN110734979A (en) | 2020-01-31 |
| CN110734979B true CN110734979B (en) | 2022-09-13 |
Family
ID=69272893
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911111701.1A Active CN110734979B (en) | 2019-11-14 | 2019-11-14 | Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110734979B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113512589B (en) * | 2021-07-14 | 2023-03-10 | 复旦大学附属中山医院 | Application of PRUNE1 gene detection in prognosis of multiple myeloma patients |
| CN115512842A (en) * | 2022-10-20 | 2022-12-23 | 南京鼓楼医院 | Intelligent webpage tool for prognosis evaluation of multiple myeloma patient |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107281170A (en) * | 2017-06-30 | 2017-10-24 | 中国人民解放军第三军医大学 | Application of the vanillic aldehyde in the medicine for preparing treatment/prevention bone metabolic disease |
-
2019
- 2019-11-14 CN CN201911111701.1A patent/CN110734979B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107281170A (en) * | 2017-06-30 | 2017-10-24 | 中国人民解放军第三军医大学 | Application of the vanillic aldehyde in the medicine for preparing treatment/prevention bone metabolic disease |
Non-Patent Citations (6)
| Title |
|---|
| A role for IL-34 in osteolytic disease of multiple myeloma;Muhammad Baghdadi等;《Blood Advances》;20190226;第3卷(第4期);541-551 * |
| Cell Fusion and Hyperactive Osteoclastogenesis in Multiple Myeloma;Franco Silvestris等;《Advances in Experimental Medicine and Biology》;20111231;第714卷;113-128 * |
| Melphalan modifies the bone microenvironment by enhancing osteoclast formation;Ryan C. Chai等;《Oncotarget》;20170710;第8卷(第40期);68047-68058 * |
| MiR-7b directly targets DC-STAMP causing suppression of NFATc1 and c-Fos signaling during osteoclast fusion and differentiation;Ce Dou等;《Biochimica et Biophysica Acta》;20140811;第1839卷(第11期);1084-1096 * |
| OC-STAMP及其在巨噬细胞系细胞融合中作用机制的研究进展;袁慧敏等;《医学综述》;20170430;第23卷(第8期);1480-1484 * |
| Sulforaphane inhibits osteoclast differentiation by suppressing the cell-cell fusion molecules DC-STAMP and OC-STAMP;Tomohiro Takagi等;《Biochemical and Biophysical Research Communications》;20161212;第483卷(第1期);718-724 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110734979A (en) | 2020-01-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2986735B1 (en) | Method for identifying the quantitative composition of blood cells in a sample | |
| Bobée et al. | Determination of molecular subtypes of diffuse large B-cell lymphoma using a reverse transcriptase multiplex ligation-dependent probe amplification classifier: a CALYM study | |
| WO2018001295A1 (en) | Molecular marker, reference gene, and application and test kit thereof, and method for constructing testing model | |
| US20180251846A1 (en) | Biomarker panel for diagnosis and prediction of graft rejection | |
| JP2009529878A (en) | Primary cell proliferation | |
| AU2013277971A1 (en) | Molecular malignancy in melanocytic lesions | |
| CA3024744A1 (en) | Methods for subtyping of lung squamous cell carcinoma | |
| KR101914348B1 (en) | Method of detecting a risk of cancer | |
| CN110734979B (en) | Application of OC-STAMP as marker for evaluating prognosis risk of multiple myeloma patient | |
| KR20170136525A (en) | Whole-blood-based mRNA marker for predicting prostate cancer and its detection method | |
| CN117233400A (en) | KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof | |
| Hashida et al. | Cutavirus Infection in Large-Plaque Parapsoriasis, a Premalignant Condition of Mycosis Fungoides | |
| EP2603604B1 (en) | Method and kit for the diagnosis and/or prognosis of tolerance in liver transplantation | |
| CN113631723B (en) | Application of tuberculosis marker in tuberculosis diagnosis and efficacy evaluation | |
| AU2019375136A1 (en) | Method for diagnosing a cancer and associated kit | |
| CN110714081B (en) | Complete set of reagent for quantitatively detecting OC-STAMP gene expression level and application thereof | |
| CN117089618A (en) | Application of AZGP1 as prognosis risk marker for patients with multiple myeloma | |
| CN117089617A (en) | Application of CADPS2 as marker in preparation of product for prognosis risk assessment of patients with multiple myeloma | |
| CN117092352A (en) | RAB3B biomarker related to multiple myeloma diagnosis and prognosis and application thereof | |
| JP6017448B2 (en) | Diagnosis method of blood diseases | |
| CN117187389A (en) | Complete reagent for quantitatively detecting KCNN3, RAB3B, CADPS2 and AZGP1 transcriptional level and application thereof | |
| JPWO2015105191A1 (en) | Method for evaluating lymphadenopathy | |
| KR20230010426A (en) | Hematologic malignancy specific markers and use thereof | |
| WO2023004460A1 (en) | Methods of detecting and/or diagnosing pancreatic cancer | |
| CN115386639A (en) | Application of RCN3 gene detection reagent in preparation of gastric cancer detection kit and gastric cancer detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230724 Address after: 100020 101, 201, 301 and 401, 1 / F, 2 / F, building 26, yard 6, Jingshun East Street, Chaoyang District, Beijing Patentee after: BEIJING JINYU MEDICAL EXAMINATION LABORATORY Co.,Ltd. Address before: 100044 No. 11 South Main Street, Xicheng District, Beijing, Xizhimen Patentee before: PEKING University PEOPLE'S HOSPITAL (PEKING UNIVERSITY SECOND CLINICAL MEDICAL College) |