CN117233400A - KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof - Google Patents
KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof Download PDFInfo
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Abstract
The application discloses a KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof. The application analyzes KCNN3 expression level in bone marrow cells of a first diagnosis and alleviation MM patient and a healthy donor by an RT-qPCR method, and discovers that the expression level of the gene in the MM patient is obviously higher than that of a normal control and possibly related to clinical course of disease. The clinical significance of this gene was subsequently explored and high expression of this gene was found to be an independent risk factor for low Progression Free Survival (PFS) in MM patients. KCNN3 has important value in diagnosis and prognosis evaluation of multiple myeloma.
Description
Technical Field
The application belongs to the technical field of medical detection, and particularly relates to a KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof.
Background
Multiple Myeloma (MM) is the second most common hematological malignancy originating from plasma cells, accounting for 10-15% of all hematological tumors. Myeloma cells proliferate maliciously in bone marrow and secrete monoclonal immunoglobulins, which can lead to organ and tissue damage. Bone marrow infiltration can result in inhibition of normal hematopoiesis and thus the appearance of anemia, infections, and bleeding symptoms. Infiltration in bone stimulates osteoclasts to enhance their osteolytic action, which can lead to bone pain, bone defects, hypercalcemia and pathological fractures. The disease is well developed for the elderly, the average age of onset is about 70 years, and the ratio of men and women is (1.6-3): 1. Current MM treatment is dominated by drug therapy. In terms of prognosis layering and efficacy evaluation, cytogenetic indexes and biochemical indexes are often relied on, and specific molecular biological markers are lacked.
KCNN3 (Potassium Calcium-Activated Channel Subfamily N Member 3), a member of the potassium-calcium activated channel subfamily N-3, encodes the product ion channel protein SK3 (small conductance calcium ion activated potassium channel). SK3 binds to Orai1 to form an SK3-Orai1 complex that is involved in calcium homeostasis regulation of tumor cells.
Disclosure of Invention
The application aims to provide a KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof.
In a first aspect, the present application claims a novel use of a substance detecting KCNN3 gene or a substance detecting mRNA encoded by KCNN3 gene or a substance detecting protein encoded by KCNN3 gene.
The present application provides an application of a substance detecting KCNN3 gene or a substance detecting mRNA encoded by KCNN3 gene or a substance detecting protein encoded by KCNN3 gene in any one of the following (1) to (6):
(1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
In a second aspect, the present application protects a novel use of KCNN3 protein or KCNN3 gene as a marker.
The application claims the application of KCNN3 protein or KCNN3 gene as a marker in any one of the following (1) - (6):
(1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
In a third aspect, the application claims a kit whose function is any one of the following 1) -6):
1) Diagnosing or aiding in the diagnosis of multiple myeloma;
2) Screening or aiding in screening patients with multiple myeloma;
3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
The kit provided by the application comprises a substance for detecting the KCNN3 gene or a substance for detecting mRNA encoded by the KCNN3 gene or a substance for detecting protein encoded by the KCNN3 gene.
In any of the above applications or kits, the substance for detecting KCNN3 gene may be a reagent and/or an instrument required for detecting the expression level of the gene by a method in the prior art, such as a reagent and/or an instrument required for detecting the expression level of the gene by high throughput sequencing, or a reagent and/or an instrument required for detecting the expression level of the gene by quantitative PCR, or a reagent and/or an instrument required for detecting the expression level of the gene by a northern hybridization technique.
The material for detecting the protein encoded by the KCNN3 gene may be a reagent and/or an instrument required for detecting the protein content using a method in the related art, such as a reagent and/or an instrument required for detecting the protein content using mass spectrometry or a related technique thereof, or a reagent and/or an instrument required for detecting the protein content using an immunohybridization technique, or a reagent and/or an instrument required for detecting the protein content using an ELISA technique, or a reagent and/or an instrument required for detecting the protein content using a protein chip or a test paper.
Further, the substance detecting the KCNN3 gene is a substance detecting the expression level of mRNA encoded by the KCNN3 gene. The substance for detecting the expression amount of the mRNA encoded by the KCNN3 gene may include a specific amplification primer and/or probe for detecting the mRNA. In one embodiment of the application, the specific amplification primers consist of a single-stranded DNA molecule shown in sequence 1 and a single-stranded DNA molecule shown in sequence 2; the probe is a single-stranded DNA molecule shown in a sequence 3. The 5' end of the probe may be labeled with a fluorescent reporter group (e.g., FAM). The 3' end of the probe may be labeled with a fluorescence quenching group (e.g., BHQ).
Still further, the expression level is a relative expression level of a reference gene of the KCNN3 gene, and specifically may be a ratio of the expression level of the KCNN3 gene to the expression level of the reference gene. The expression amount of the KCNN3 gene and the expression amount of the reference gene may be copy numbers obtained from standard curves and CT values.
Further, the substance for detecting the expression amount of mRNA encoded by the KCNN3 gene may further comprise a specific amplification primer and/or probe for detecting the expression amount of mRNA of the reference gene. The reference gene may specifically be the ABL1 gene. In one embodiment of the present application, the specific amplification primers for detecting the ABL1 gene mRNA consist of a single-stranded DNA molecule shown in sequence 4 and a single-stranded DNA molecule shown in sequence 5; the probe for detecting ABL1 gene mRNA is a single-stranded DNA molecule shown in a sequence 6, wherein the 5 '-end of the probe can be marked with a fluorescence reporting group (such as FAM), and the 3' -end of the single-stranded probe can be marked with a fluorescence quenching group (such as BHQ).
The kit can also comprise a data processing device A; a module is arranged in the data processing device A; the module has the following functions: diagnosing whether the tester is a multiple myeloma patient according to the relative expression quantity of KCNN3 in the bone marrow mononuclear cells of the tester: if the relative expression amount of KCNN3 in the bone marrow mononuclear cells of the tested person is larger than that of the healthy control person, the tested person is or is suspected to be a patient with multiple myeloma; otherwise, the subject is not or suspected of not being a multiple myeloma patient.
The kit may further comprise a data processing device B; a module is arranged in the data processing device B; the module has the following functions: assessing the prognosis risk or progression-free survival rate or progression-free survival time of a multiple myeloma patient based on the relative expression of KCNN3 in bone marrow mononuclear cells of the multiple myeloma patient: the risk of prognosis for patients with multiple myeloma in the KCNN3 low expression group is lower than or candidates for patients with multiple myeloma in the KCNN3 high expression group; progression free survival of multiple myeloma patients in KCNN3 low expression group is higher than or candidates for multiple myeloma patients in KCNN3 high expression group; the progression free survival time of the patients with multiple myeloma in the KCNN3 low expression group is longer than or candidates for patients with multiple myeloma longer than that in the KCNN3 high expression group.
The kit may further comprise a data processing device C; the data processing device C is internally provided with a module; the module has the following functions: assessing the disease course progression of a multiple myeloma patient based on the relative expression level of KCNN3 in bone marrow mononuclear cells of the multiple myeloma patient: the risk of relapse in patients with multiple myeloma in KCNN3 low expression group is lower than or candidates for patients with multiple myeloma in KCNN3 high expression group.
The above KCNN3 high expression group and KCNN3 low expression group can be judged as follows: taking isolated bone marrow of a group to be tested consisting of a plurality of patients with multiple myeloma without any treatment measures as a sample, measuring the relative expression quantity of KCNN3 in each sample, arranging and halving the group to be tested according to the sequence of the relative expression quantity from low to high, taking 1/2 of the group to be tested with low expression quantity as a KCNN3 low expression group, and taking the rest 1/2 of the groups to be tested as a KCNN3 high expression group.
In any of the above applications or kits, the nucleotide sequence of the KCNN3 gene is as shown in sequence 9.
The amino acid sequence of the KCNN3 protein is shown as a sequence 10.
In the present application, the prognostic risk is a prognostic risk of a patient with primary diagnosis of multiple myeloma. The prognosis 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 risk of prognosis; the higher the progression free survival, the lower the risk of prognosis.
The disease progression (recurrence risk) refers to the recurrence risk of patients with multiple myeloma after remission.
In any of the above applications or kits, the screening or aiding in screening of multiple myeloma patients includes screening or aiding in screening of primary multiple myeloma patients and screening or aiding in screening of relapsing multiple myeloma patients.
In any of the above applications or kits, the multiple myeloma is adult multiple myeloma.
The application analyzes KCNN3 expression level in bone marrow cells of a first diagnosis and alleviation MM patient and a healthy donor by an RT-qPCR method, and discovers that the expression level of the gene in the MM patient is obviously higher than that of a normal control and possibly related to clinical course of disease. The clinical significance of this gene was subsequently explored and high expression of this gene was found to be an independent risk factor for low Progression Free Survival (PFS) in MM patients. KCNN3 has important value in diagnosis and prognosis evaluation of multiple myeloma.
Drawings
FIG. 1 shows KCNN3 expression levels in bone marrow cells of a primary diagnosis and remission MM patient and a healthy donor. Lines represent median and quartile values; * P < 0.0001.
FIG. 2 is a ROC graph of KCNN3 diagnostic first diagnosis MM.
FIG. 3 is a graph showing the relationship between KCNN3 expression level and the clinical course of MM. KCNN3 expression levels in bone marrow single nuclear cell specimens of initial diagnosis, remission and disease progression in 6 MM patients.
FIG. 4 shows the relationship between KCNN3 expression level and the progression-free survival rate of MM prognosis.
Detailed Description
The following detailed description of the application is provided in connection with the accompanying drawings that are presented to illustrate the application and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the application in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The nucleotide sequence of the KCNN3 gene is shown as a sequence 9 in a sequence table, and the KCNN3 protein shown as a coding sequence 10.
Example 1, application of KCNN3 as biomarker in diagnosis and prognosis evaluation of multiple myeloma
1. Study object and method
1. Study object
Bone marrow specimens collected from 145 cases of primary patients with multiple myeloma from the institute of hematopathy in the civil hospital at university of Beijing during the period of 1 month to 12 months in 2014 were taken as subjects. 145 multiple myeloma patients included 92 men, 53 women, with a median age of 60 years and an age ranging from 36 years to 87 years, and were followed until death, no follow-up, or 8 months of 2019. The diagnostic criteria for multiple myeloma refer to guidelines of the national integrated cancer network (NCCN), and the staging is performed according to the Durie-Salmon (D-S) staging system, the International Staging System (ISS) and the revised International staging system (R-ISS). Treatment and efficacy assessments for patients with multiple myeloma are made with reference to the guidelines above. Progression Free Survival (PFS) is defined as the time from the onset of primary treatment to the first occurrence of disease progression, which is an event.
In addition, 87 total bone marrow specimens were collected during follow-up of 87 primary patients with multiple myeloma, including 24 bone marrow specimens that were treated to obtain Complete Remission (CR), 27 bone marrow specimens that were treated to obtain Very Good Partial Remission (VGPR), 32 bone marrow specimens that were treated to obtain Partial Remission (PR), and 4 bone marrow specimens that were treated to develop disease progression. Normal control bone marrow specimens were obtained from adult healthy volunteers in 46 total.
Specimens used in this study protocol were approved by the ethical committee of the civil hospital at the university of Beijing. All healthy volunteers and patients signed informed consent.
2. Bone marrow mononuclear cell extraction and RT-qPCR
Mononuclear cells in bone marrow specimens were separated using Ficoll lymphocyte separation medium and density gradient centrifugation, and RNA was extracted and reverse transcribed into cDNA. And (3) taking cDNA as a template, and performing RT-qPCR by adopting a KCNN3 primer pair (wherein the KCNN3 primer pair consists of a KCNN3 upstream primer and a KCNN3 downstream primer, the size of an amplified product is 106bp, and the nucleotide sequence is shown as a sequence 7 in a sequence table) and a KCNN3 probe. And (3) taking cDNA as a template, and performing RT-qPCR by using an ABL1 primer pair (the ABL1 primer pair consists of an ABL1 upstream primer and an ABL1 downstream primer, the size of an amplified product is 124bp, and a nucleotide sequence is shown as a sequence 8 in a sequence table) and an ABL1 probe. The following 10 μl PCR reaction system was configured using PCR Master Mix kit: 5 mu LUniversal PCR Master Mix; upstream primer 0.9. Mu.M, downstream primer 0.9. Mu.M, probe 0.25. Mu.M; 150-500ng cDNA, primer sequences and fluorescent probe sequences are shown in Table 1. qPCR was performed using an ABI 7500FAST PCR amplification apparatus under the following reaction conditions: 50℃2min,95℃10min, then 95℃15s,60℃1min for 40 cycles. The copy numbers of KCNN3 and ABL1 were calculated by standard curve method using ABL1 as an internal reference. Serial dilutions of ABL1 expressing plasmids (see "Gabert J, beillillard E, van der Velden VH, bi W, grimwade D, pallisgaard)N, et al, standard and quality control studies of, "real-time" quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia-a Europe Against Cancer program Leukemia 2003;17:2318-2357, "ABL1 plasma" in the text) (10 6 、10 5 、10 4 、10 3 、10 2 、10 1 And 10 0 Copy/. Mu.L) and KCNN3 positive bone marrow samples were used to construct a standard quantification curve. The curve threshold is set to 0.082. Obtaining Ct values of the ABL1 and KCNN3 through amplification curves of sample reference genes ABL1 and KCNN3 genes and a set threshold (0.082), obtaining copy numbers of the samples ABL1 and KCNN3 according to an ABL1 standard curve (which is calculated by referring to the ABL1 standard curve for reducing experimental errors because the KCNN3 and the ABL1 amplification efficiency are similar), dividing the copy number of the KCNN3 by the copy number of the ABL1 to obtain the expression quantity of the sample KCNN3, multiplying the result by one hundred percent for consistency with the form reported by clinical routine, and finally representing the copy number of the KCNN 3/the copy number multiplied by the percentage of the ABL1 = the expression quantity of the sample KCNN 3.
TABLE 1 KCNN3 and ABL1 primers and probe sequences
| Primer(s) | Sequence (5 '-3') |
| KCNN3 upstream primer | 5'-CAGCTCACCAAGCGGATCA-3' (sequence 1) |
| KCNN3 downstream primer | 5'-TCACTTTGGCATGGTCAATCTT-3' (sequence 2) |
| KCNN3 probe | 5'-FAM-TGCTGCAGCCAATGTCCTTCGG-BHQ-3' (sequence 3) |
| ABL1 upstream primer | 5'-TGGAGATAACACTCTAAGCATAACTAAAGGT-3' (sequence 4) |
| ABL1 downstream primer | 5'-GATGTAGTTGCTTGGGACCCA-3' (sequence 5) |
| ABL1 probe | 5'-FAM-CCATTTTTGGTTTGGGCTTCACACCATT-BHQ-3' (sequence 6) |
3. Statistical analysis
Statistical analysis was performed using SPSS22.0, graphpad Prism 7. And comparing the difference of the two groups of data, wherein the classification variable data adopts chi-square test, the continuous variable data adopts t test, and the difference is less than 0.05, so that the statistical significance is realized. Subject work (receiver operating characteristic, ROC) curves were used to evaluate the specificity and sensitivity of diagnostic indicators. The Jooden Index (Youden Index) is used to calculate the cut-off value for the diagnosis. Survival analysis was performed using Kaplan-Meier using log-rank test. The Cox proportional hazards regression model performs multi-factor analysis. The variable with P more than 0.1 is gradually removed by a back-off method, and P less than 0.05 has statistical significance.
2. Results of the study
1. Expression level of KCNN3 in multiple myeloma
The KCNN3 expression levels were analyzed for 145 bone marrow specimens from primary multiple myeloma patients, 83 bone marrow specimens from multiple myeloma remission patients, and 46 bone marrow specimens from healthy donor controls.
The results show that the expression level of KCNN3 in bone marrow cells of primary patients with multiple myeloma (median 306.6%; range 11.68-15624.15%) is significantly higher than that of patients with remission (median 6.07%; range 0.36-835.58%; P < 0.0001) and healthy donor controls (median 3.49%; range 0.44-14.37%; P < 0.0001). (FIG. 1).
ROC curve analysis showed that the area under the curve (area under the curve, AUC) of KCNN3 diagnostic MM was 0.9996 (95% confidence interval 0.9984-1; p < 0.0001; fig. 2), with a maximum approximate sign of KCNN3 expression level of 18.21%. With this as the diagnostic threshold for MM, the overexpression rate of KCNN3 in the initial MM was 97.931%.
2. KCNN3 expression level is correlated with clinical course of multiple myeloma
To further investigate the relationship between KCNN3 and the clinical course of adult multiple myeloma, analysis of KCNN3 expression levels was performed on bone marrow specimens at the time of initial diagnosis, complete remission and disease progression in 6 adult multiple myeloma patients.
The results showed that the levels of KCNN3 expression in bone marrow cells were significantly reduced at complete remission compared to the initial diagnosis of bone marrow cells, whereas the levels of KCNN3 expression in bone marrow cells were significantly increased at disease progression (recurrence) compared to complete remission (fig. 3).
3. Relationship between KCNN3 expression level and general clinical characteristics of multiple myeloma
145 patients initially diagnosed with multiple myeloma are classified into a KCNN3 high-expression group and a KCNN3 low-expression group according to the median value of KCNN3 expression levels as follows: measuring the relative expression quantity of KCNN3 in each patient bone marrow specimen, then arranging and halving the groups to be tested according to the sequence from low to high of the relative expression quantity of KCNN3, taking 1/2 of the groups to be tested with low relative expression quantity as KCNN3 low expression groups, taking the rest 1/2 of the groups to be tested as KCNN3 high expression groups, and analyzing the relation between the expression level of KCNN3 and the general clinical characteristics of multiple myeloma.
The results showed that KCNN3 high-expressing group patients correlated with higher levels of bone marrow plasma cell ratio (p=0.001), more prone to high-risk cytogenetic markers associated with poor prognosis (P < 0.001) (table 2).
TABLE 2 relationship between KCNN3 expression levels and general clinical characteristics of adult multiple myeloma in initial diagnosis
4. Relationship between KCNN3 expression level and prognosis of patients with multiple myeloma
Survival analysis was performed on 118 patients with multiple myeloma with prognostic information by Kaplan-Meier method, and according to the above step 3, the patients were divided into two groups (KCNN 3 high-expression group and KCNN3 low-expression group) according to the relative expression amount of KCNN3, and it was found that the 48 month progression-free survival rate (PFS) was 9.43% and 25.62% in the KCNN3 high-expression group (n=60) patient and KCNN3 low-expression group (n=58) patient, respectively, and the KCNN3 high-expression group patient had a shorter progression-free survival time than the KCNN3 low-expression group patient (median 17.7vs.28.17 months; p=0.0077; fig. 4). Since there are many factors affecting MM prognosis, to further clarify the effect of KCNN3 on prognosis, the following factors were included in the multi-factor analysis: beta 2 microglobulin level (more than or equal to vs. < 3.5 mg/L) at the initial diagnosis, lactate dehydrogenase level (more than or equal to vs. < 245 g/L) at the initial diagnosis), and hematopoietic stem cell transplantation (yes vs. no). The results show that KCNN3 high expression is an independent risk factor for progression free survival (table 3).
TABLE 3 multifactorial analysis of Progression Free Survival (PFS) in adult multiple myeloma patients
| End of the year | Risk ratio (95% confidence interval) | P value |
| KCNN3: high expression vs. low expression | 1.698(1.087-2.651) | 0.020* |
| Beta 2 microglobulin level: not less than 3.5mg/L vs.<3.5mg/L | 1.360(0.773-2.391) | 0.286 |
| Lactate dehydrogenase level:<245g/L vs.≥245g/L | 1.052(0.620-1.785) | 0.850 |
| receiving hematopoietic stem cell transplantation: is vs. no | 0.814(0.455-1.455) | 0.487 |
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (10)
1. Use of a substance detecting KCNN3 gene or a substance detecting mRNA encoded by KCNN3 gene or a substance detecting protein encoded by KCNN3 gene in any one of the following (1) to (6):
(1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
2. The use according to claim 1, characterized in that: the substance for detecting the mRNA encoded by the KCNN3 gene comprises a reagent and/or an instrument for detecting the expression amount of the mRNA encoded by the KCNN3 gene in the bone marrow mononuclear cells.
3. The use according to claim 2, characterized in that: the reagent for detecting the expression level of the mRNA encoded by the KCNN3 gene in the bone marrow mononuclear cells comprises a specific amplification primer and/or a probe for detecting the mRNA encoded by the KCNN3 gene.
4. A use according to claim 3, characterized in that: the specific amplification primer consists of a single-stranded DNA molecule shown in a sequence 1 and a single-stranded DNA molecule shown in a sequence 2;
the probe is a single-stranded DNA molecule shown in a sequence 3.
5. A kit comprising a substance that detects KCNN3 gene or a substance that detects mRNA encoded by KCNN3 gene or a substance that detects protein encoded by KCNN3 gene; the function of the kit is any one of the following 1) -6):
1) Diagnosing or aiding in the diagnosis of multiple myeloma;
2) Screening or aiding in screening patients with multiple myeloma;
3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
6. The kit of claim 5, wherein: the substance for detecting the mRNA encoded by the KCNN3 gene comprises a reagent and/or an instrument for detecting the expression amount of the mRNA encoded by the KCNN3 gene in the bone marrow mononuclear cells.
7. The use according to claim 6, characterized in that: the reagent for detecting the expression level of the mRNA encoded by the KCNN3 gene in the bone marrow mononuclear cells comprises a specific amplification primer and/or a probe for detecting the mRNA encoded by the KCNN3 gene.
8. The use according to claim 7, characterized in that: the specific amplification primer consists of a single-stranded DNA molecule shown in a sequence 1 and a single-stranded DNA molecule shown in a sequence 2;
the probe is a single-stranded DNA molecule shown in a sequence 3.
Use of KCNN3 protein or KCNN3 gene as a marker in any of the following (1) - (6):
(1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
10. The use according to any one of claims 1 to 4 or the kit according to any one of claims 5 to 8 or the use according to claim 9, characterized in that: the nucleotide sequence of the KCNN3 gene is shown as a sequence 9; the amino acid sequence of the KCNN3 protein is shown as a sequence 10.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311089449.5A CN117233400A (en) | 2023-08-28 | 2023-08-28 | KCNN3 gene detection kit for diagnosis and prognosis evaluation of multiple myeloma and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480645A (en) * | 2022-01-13 | 2022-05-13 | 上海交通大学医学院附属仁济医院 | Multiple myeloma exhausted NK cell subgroup, characteristic gene and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114480645A (en) * | 2022-01-13 | 2022-05-13 | 上海交通大学医学院附属仁济医院 | Multiple myeloma exhausted NK cell subgroup, characteristic gene and application thereof |
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