WO2021000673A1 - Application de rbm46 en tant que marqueur tumoral testiculaire - Google Patents

Application de rbm46 en tant que marqueur tumoral testiculaire Download PDF

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WO2021000673A1
WO2021000673A1 PCT/CN2020/093005 CN2020093005W WO2021000673A1 WO 2021000673 A1 WO2021000673 A1 WO 2021000673A1 CN 2020093005 W CN2020093005 W CN 2020093005W WO 2021000673 A1 WO2021000673 A1 WO 2021000673A1
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rbm46
protein
detection agent
mrna
quantitative detection
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PCT/CN2020/093005
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刘年
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北京太东生物科技有限公司
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    • 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
    • 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
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present disclosure relates to the field of medical diagnosis, in particular to the application of Rbm46 as a marker for testicular tumors.
  • Testicular tumors are one of the more common tumors in the urinary system. Most of the patients are males in young adults, almost all of them are malignant. Testicular cancer accounts for 1% of systemic malignancies and 3%-9% of genitourinary system malignancies. The incidence of testicular cancer differs between races and regions. The incidence of testicular cancer in whites is higher than that of non-whites. The incidence of whites in the United States is about 6/100,000, and the incidence in Africa is 1/100,000, and the incidence is increasing year by year. Testicular tumors are divided into primary and secondary types. Most of them are primary, and secondary is extremely rare.
  • Testicular tumors are almost always malignant, with germ cell tumors accounting for 90%-95% and non-germ cell tumors accounting for 5%-10%.
  • seminoma is the most common, accounting for about 40% to 50% of primary testicular tumors, followed by embryonic cancer, about 20% to 30%, and teratoma, about 10%.
  • testicular tumors of other cell types are rare.
  • the cause of the disease is still unknown, and it is currently believed that the disease is related to heredity and acquired factors. Among them, it is most closely related to cryptorchidism.
  • Cryptorchidism is 10 to 14 times more likely than normal people to develop tumors.
  • Intra-abdominal cryptorchidism is higher than that of groin. Orchiopexy does not reduce the incidence of malignancy, but it can make tumors more vulnerable Find.
  • testicular cancer has no obvious clinical symptoms, and the initial misdiagnosis rate of testicular cancer is as high as 25%.
  • the existing imaging studies cannot find it yet.
  • tumor markers with sensitivity and specificity so it is of great clinical significance to find testicular cancer markers with higher specificity.
  • the present disclosure relates to the application of a quantitative detection agent for Rbm46 mRNA or protein in the preparation of reagents or kits for the diagnosis and/or prognosis evaluation of testicular tumors, wherein the reduced expression of Rbm46 mRNA or protein is an indication and/or of testicular tumors Or an indication of poor prognosis for testicular tumors.
  • testicular tumors can be benign or malignant, such as advanced or metastatic malignant testicular tumors.
  • Rbm46 mRNA or protein is of animal origin, preferably primate, more preferably human.
  • the quantitative detection agent for Rbm46 mRNA includes a reagent suitable for at least one of the following methods:
  • the quantitative detection agent for Rbm46 mRNA is a probe or primer that can specifically bind to Rbm46 mRNA or Rbm46 cDNA.
  • the probe or primer bears a detectable label.
  • the quantitative detection agent for Rbm46 protein is a reagent required to specifically measure Rbm46 protein.
  • the quantitative detection agent for Rbm46 protein includes one or more of lectin, aptamer antibody, or antibody fragment capable of specifically binding to Rbm46 protein.
  • the quantitative detection agent for Rbm46 protein has an affinity of at least 10 7 l/mol for its corresponding target molecule, preferably an affinity of 10 8 l/mol, or more preferably 10 9 l/mol for its target molecule .
  • the binding affinity of the quantitative detection agent for Rbm46 protein and biomolecules other than the Rbm46 protein is at most only 10% or less, only 5% or less of the affinity with the target molecule, Only 2% or less, or only 1% or less.
  • antibodies or antibody fragments can be obtained by immunizing Rbm46 protein or fragments thereof as an antigen.
  • the immunized subjects can be selected to include humans and all domesticated animals (such as domestic animals and pets) and wild animals and birds.
  • the antibody or antibody fragment is labeled with an indicator that shows signal strength.
  • the detection agent capable of specifically binding to the Rbm46 polypeptide is an antibody or antibody fragment.
  • the present disclosure also relates to a qRT-PCR primer for Rbm46 mRNA, the upstream primer of which is shown in SEQ ID NO: 1, and the downstream primer is shown in SEQ ID NO: 2.
  • the present disclosure also relates to a kit for detecting the expression level of Rbm46 mRNA, which includes the primers described above.
  • the kit further includes one or more of reverse transcription reagents, internal reference primers, fluorescent substances for indicating the amount of DNA synthesis, qPCR reaction buffer, dNTPs, DNA polymerase, and water.
  • the internal reference primer is selected from primers of GAPDH, tubulin or actin; water is nuclease-free water, such as reverse osmosis water, distilled water or deionized water; DNA polymerase is selected from Taq, Bst, Vent, Phi29, Pfu , Tru, Tth, Tl1, Tac, Tne, Tma, Tih, Tf1, Pwo, Kod, Sac, Sso, Poc, Pab, Mth, Pho, ES4 DNA polymerase, Klenow fragment.
  • the kit further includes detection reagents for one or more of serum ⁇ -HCG, AFP, and LDH.
  • the present disclosure also relates to a method for evaluating testicular tumors, the method comprising:
  • step (a) measuring the expression level of Rbm46 mRNA or protein in the sample, (b) optionally, measuring the concentration of one or more other testicular tumor markers in the sample, and (c) using step (a)
  • the measurement results of and optional step (b) are used to evaluate testicular cancer, where the decreased expression of Rbm46 mRNA or protein is an indication of testicular cancer.
  • the measurement sample includes blood (whole blood), serum, plasma, cell culture supernatant, saliva, cerebrospinal fluid, semen, prostatic fluid, tissue, or tissue lysate.
  • Rbm46 mRNA or protein is highly expressed in normal cells and tissues of the testis, while it is significantly low expressed in testicular cancer cells.
  • the expression level is significantly correlated with clinicopathological parameters, and has important clinical application value.
  • Figure 1 shows the expression of Rbm46 mRNA in testicular cancer tumors in an embodiment of the present disclosure
  • FIG. 1 The expression of Rbm46 protein in testicular cancer tumors in an embodiment of the present disclosure; wherein 1#, 2#, and 3# represent different sample numbers.
  • the present disclosure relates to a quantitative detection agent for Rbm46 mRNA or protein, which is used for the diagnosis and/or prognosis evaluation of testicular tumors, wherein the decreased expression of Rbm46 mRNA or protein is an indication of testicular tumors and/or an indication of poor prognosis of testicular tumors.
  • the present disclosure relates to the application of a quantitative detection agent for Rbm46 mRNA or protein in the preparation of reagents or kits for the diagnosis and/or prognosis evaluation of testicular tumors, wherein the reduced expression of Rbm46 mRNA or protein is an indication and/or of testicular tumors Or an indication of poor prognosis for testicular tumors.
  • prognosis is the prediction of disease, particularly testicular tumors, including whether symptoms and signs will improve or worsen over time (and how quickly) or remain stable; expectations for quality of life, such as The ability to carry out daily activities; the likelihood of complications and related health problems; and the likelihood of survival (including life expectancy).
  • diagnosis is the process of determining which disease or condition can explain the symptoms and signs of an individual.
  • the information needed for diagnosis is usually collected from the medical history and physical examination of the individual seeking medical care.
  • Rbm46 mRNA or protein provides a new marker for prognostic evaluation and judgment of testicular tumors: Rbm46 mRNA or protein.
  • Clinical studies have confirmed that its expression in testicular tumors is lower than that of normal tissues. Patients with low levels of Rbm46 mRNA or protein in cancer tissues have shorter survival periods and higher mortality. According to this, Rbm46 mRNA or protein as a biomarker effectively improves the positive rate of testicular tumor prognosis using kits in clinic.
  • testicular tumors can be benign or malignant, such as advanced or metastatic malignant testicular tumors.
  • Testicular tumors can also be replaced by the term "testicular cancer”.
  • advanced or metastatic malignant testicular tumor refers to an advanced, unresectable and/or metastatic relapsed or refractory malignant testicular tumor that is diagnosed histologically or cytologically, which is ineffective or non-existent to standard therapies. Effective treatment for it.
  • the Rbm46 protein is RNA binding motif protein 46, which is often regarded as cancer-testis antigens (CTAs) in the prior art. CTAs have specific normal expression in gamete tissues and abnormal expression in cancer. However, the present disclosure has unexpectedly discovered that the expression of Rbm46 mRNA or protein in testicular tumors and adjacent tissues is significantly different, suggesting that Rbm46 mRNA or protein can be used as a marker for diagnosis and/or prognosis evaluation of testicular cancer.
  • CTAs cancer-testis antigens
  • Rbm46 mRNA or protein is of animal origin, preferably primate, more preferably human.
  • the term "quantitative detection agent for Rbm46 mRNA" in the present disclosure should not only be understood as a detection agent for Rbm46 mRNA, but should include other detection reagents known to those skilled in the art that can reflect the expression level of Rbm46 mRNA.
  • the expression level of Rbm46 mRNA can be indirectly detected by quantitatively detecting the cDNA obtained by reverse transcription of Rbm46 mRNA.
  • Rbm46 mRNA and Rbm46 protein can be referred to as “markers”, “marker genes” (specifically Rbm46 mRNA), “biochemical markers”, and “marker polypeptides” ( Specifically refers to Rbm46 protein), “testicular tumor marker” or “testicular cancer marker”. It specifically refers to a molecule to be used as a target for the analysis of a patient's experimental sample.
  • the Rbm46 mRNA used as a marker in the present disclosure is expected to include its full-length ribonucleotide sequence, or naturally-occurring variants, or full-length sequences and fragments of variants, especially fragments whose specific sequences can be detected and determined , More preferably a fragment that can be distinguished from other RNA sequences in testicular tissue. It preferably comprises at least 7, 8, 9, 10, 11, 12, 15 or 20 consecutive ribonucleotides of the full-length ribonucleotide sequence.
  • the Rbm46 protein used as a marker in the present disclosure is expected to include naturally-occurring variants of the protein and fragments of the protein or the variants, particularly immunologically detectable fragments.
  • the immunologically detectable fragment preferably comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 15 or 20 consecutive amino acids of the marker polypeptide.
  • the expression "Rbm46 protein” includes the complete protein sequence of Rbm46 and the marker polypeptide defined above.
  • ribonucleotides/proteins/polypeptides released by cells or ribonucleotides present in the extracellular matrix may be damaged (for example, during inflammation), and may be degraded or Cut into fragments like this.
  • mRNA, protein or fragments thereof may also be present as part of the complex.
  • Such a complex can also be used as a marker in the sense of the present disclosure.
  • the marker polypeptide or variants thereof may carry post-translational modifications.
  • post-translational modifications are glycosylation, acylation and/or phosphorylation.
  • “Naturally occurring variants” should be understood to mean that higher animal genes are usually accompanied by high frequency polymorphisms. There are also many molecules that produce isotypes containing mutually different amino acid sequences during the splicing process. Any gene associated with cancer-related diseases that has an activity similar to that of the marker gene is included in the marker gene, even if it has a nucleotide sequence difference due to polymorphism or isotype.
  • RNA quantitative detection reagents can be selected from reagents known to those skilled in the art, such as nucleic acids that can hybridize to the RNA and labeled with fluorescent labels; in common cases, RNA detection reagents can be selected from RT-PCR primers, and Primers for the amplification of RT-PCR products-cDNA;
  • the quantitative detection agent for Rbm46 mRNA includes a reagent suitable for at least one of the following methods:
  • the quantitative detection agent for Rbm46 mRNA is a probe or primer that can specifically bind to Rbm46 mRNA or Rbm46 cDNA.
  • the marker gene may include homologues of other species except human. Therefore, unless explicitly stated, the expression “marker gene” refers to a homolog of a marker gene unique to a species or a foreign marker gene that has been introduced into an individual. Similarly, it should be understood that "a homolog of a marker gene” can be used as a probe to hybridize to a human marker gene under stringent conditions. Such stringent conditions are known to those skilled in the art (who can select appropriate conditions to produce the same stringency through experiment or experience).
  • a polynucleotide containing the nucleotide sequence of the marker gene or the complementary strand of the nucleotide sequence of the marker gene and having a nucleotide sequence of at least 15 nucleotides can be used as a primer or a probe. Therefore, the "complementary strand” refers to one strand of double-stranded DNA composed of A:T (U for RNA) and G:C base pairs relative to the other strand.
  • complementary means not only a sequence that is completely complementary to a region of at least 15 consecutive nucleotides, but also a sequence having at least 40% in some cases, 50% in some cases, and 60% in some cases. %, 70% in some cases, 80% in some cases, 90% in some cases, and in some cases 95% or higher nucleotide sequence homology. The degree of homology between nucleotide sequences can be determined using an algorithm such as BLAST.
  • polynucleotides can be used as probes for detecting marker genes or as primers for amplifying marker genes.
  • polynucleotides When used as primers, polynucleotides generally comprise 15 bp to 100 bp, and in certain embodiments 15 bp to 35 bp of nucleotides.
  • DNA When used as a probe, DNA contains the entire nucleotide sequence of the marker gene (or its complementary strand), or a partial sequence thereof having at least 15 bp nucleotides.
  • the 3'region must be complementary to the marker gene, and the 5'region can be linked to a restriction endonuclease recognition sequence or tag.
  • Polynucleotide can be DNA or RNA.
  • oligonucleotide means a polynucleotide having a relatively low degree of polymerization. Oligonucleotides are also included in polynucleotides.
  • Northern hybridization dot blot hybridization, or DNA microarray technology can be used for detection of disorders and/or diseases using hybridization technology.
  • gene amplification techniques such as the RT-PCR method can be used. By using the PCR amplification monitoring method in the gene amplification step of RT-PCR, the expression of marker genes can be analyzed more quantitatively.
  • the probe or primer has a detectable label.
  • the detection target (reverse transcript of DNA or RNA) is hybridized with a probe labeled with a fluorescent dye and a quencher that absorbs fluorescence.
  • a probe labeled with a fluorescent dye and a quencher that absorbs fluorescence.
  • Taq polymerase degrades the probe with its 5'-3' exonuclease activity
  • the fluorescent dye and the quencher are separated from each other, thereby detecting fluorescence.
  • Real-time detection of fluorescence By simultaneously measuring a standard sample in which the copy number of the target is known, the cycle number (in which PCR amplification is linear) can be used to determine the copy number of the target in the subject sample.
  • the cycle number in which PCR amplification is linear
  • the PCR amplification monitoring method can be performed using any appropriate method.
  • the quantitative detection agent for Rbm46 protein is a reagent required to specifically measure Rbm46 protein.
  • the reagents (specific binding agents) required to specifically measure Rbm46 protein are, for example, ligands or receptors for Rbm46 protein (if present), lectins that bind to Rbm46 protein, and Rbm46 binding agents. Protein aptamers or antibodies and antibody fragments that bind to Rbm46 protein. Specific binding agent having an affinity of at least 10 7 l / mol for its corresponding target molecule. Specific binding agent to its target molecule is preferably 10 8 l / mol, or more preferably 10 9 l / mol affinity. The skilled person will understand that using the term "specific" means that other biomolecules present in the sample do not significantly bind to the specific binding agent of the Rbm46 protein.
  • the level of binding to biomolecules other than the target molecule produces a binding affinity that is at most only 10% or less, only 5% or less, and only 2% of the affinity to the target molecule, respectively. % Or less, or only 1% or less.
  • the preferred specific binding agent will simultaneously meet the above minimum criteria for affinity and specificity.
  • antibody includes polyclonal antibodies and monoclonal antibodies, and the term “antibody fragment” includes antigen compound binding fragments of these antibodies, including Fab, F(ab') 2 , Fd, Fv, scFv, bispecific antibodies and antibodies The smallest recognition unit, and single-chain derivatives of these antibodies and fragments, such as scFv-Fc, etc.
  • the type of antibody can choose IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, IgD.
  • antibody includes naturally-occurring antibodies and non-naturally-occurring antibodies, including, for example, chimeric, bifunctional, humanized, and human antibodies, and related synthetic antibodies. Isoforms.
  • antibody can be used interchangeably with "immunoglobulin”.
  • the antibodies or antibody fragments can be obtained by immunizing Rbm46 protein or fragments thereof as antigens.
  • the immunized subjects can be selected including humans and all livestock (such as domestic animals and pets) and wild animals and birds. Limitations include cattle, horses, dairy cows, pigs, sheep, goats, rats, mice, dogs, cats, rabbits, camels, donkeys, deer, minks, chickens, ducks, geese, turkeys, cockfighting, etc.
  • the antibody or antibody fragment is labeled with an indicator that shows signal strength.
  • the present disclosure also relates to a qRT-PCR primer for Rbm46 mRNA, the upstream primer of which is shown in SEQ ID NO: 1, and the downstream primer is shown in SEQ ID NO: 2.
  • the primer can be used for the diagnosis and/or prognosis evaluation of human testicular cancer.
  • the present disclosure also relates to a kit for detecting the expression level of Rbm46 mRNA, which includes the primers described above.
  • the kit can be used for the diagnosis and/or prognosis evaluation of human testicular cancer.
  • the kit further includes one or more of reverse transcription reagents, internal reference primers, fluorescent substances for indicating the amount of DNA synthesis, qPCR reaction buffer, dNTPs, DNA polymerase, and water.
  • the internal reference primer is selected from primers of GAPDH, tubulin or actin.
  • the water is nuclease-free water, such as reverse osmosis water, distilled water, and deionized water.
  • the DNA polymerase is selected from Taq, Bst, Vent, Phi29, Pfu, Tru, Tth, T11, Tac, Tne, Tma, Tih, Tf1, Pwo, Kod, Sac, Sso, Poc, Pab , Mth, Pho, ES4 DNA polymerase, Klenow fragment.
  • the kit further includes detection reagents for one or more of serum ⁇ -HCG, AFP and LDH.
  • Elevated serum AFP, HCG, and LDH concentrations indicate testicular cancer or its poor prognosis.
  • High serum concentrations of HCG are a powerful factor for poor prognosis. If the concentration continues to increase, the risk of recurrence increases accordingly.
  • the International Germ Cell Tumor Cooperation Group (IGCCCG) has used the combined detection of serum HCG, AFP, and LDH concentrations as the basis for the classification of metastatic germ cell tumors. According to the concentration of markers, tumors can be classified into good prognosis, fair prognosis, and poor prognosis. Primary tumor and lung metastasis.
  • the Rbm46 mRNA/protein provided in the present disclosure can be used in combination with the above-mentioned molecular markers to detect testicular tumors more accurately.
  • the present disclosure also relates to a method for evaluating testicular tumors, the method comprising:
  • step (a) measuring the expression level of Rbm46 mRNA or protein in the sample, (b) optionally, measuring the concentration of one or more other testicular tumor markers in the sample, and (c) using step (a)
  • the measurement results of and optional step (b) are used to evaluate testicular cancer, where the decreased expression of Rbm46 mRNA or protein is an indication of testicular cancer.
  • step a) in step a), the above-mentioned reagents required to specifically quantitatively measure Rbm46 mRNA or protein are used for measurement.
  • the ideal scenario for diagnosis is a situation where a single event or process can cause various diseases, for example, in infectious diseases. In all other cases, the correct diagnosis can be very difficult, especially when the etiology of the disease is not fully understood.
  • a diagnosis without biochemical markers is 100% specific and 100% sensitive.
  • the determination of whether the subject sample has a testicular tumor compared with the normal control sample can be performed by statistical methods known in the art, and the confidence interval and/or p-value are used for confirmation.
  • the confidence interval is 90%, 95%, 97.5%, 98%, 99%, 99.5%, 99.9%, or 99.99% and the p-value is 0.1, 0.05, 0.025, 0.02, 0.01, 0.005, 0.001 or 0.0001.
  • biochemical markers for example, serum AFP, HCG, LDH, or Rbm46 mRNA or protein confirmed in the present disclosure
  • serum AFP, HCG, LDH, or Rbm46 mRNA or protein confirmed in the present disclosure can be used to evaluate, for example, the existence or severity of a disease with a certain probability or predictive value. Therefore, in routine clinical diagnosis, various clinical symptoms and biological markers are usually considered to diagnose, treat and control underlying diseases.
  • the method is used for prognostic evaluation of testicular tumors.
  • the measurement sample includes blood (whole blood), serum, plasma, cell culture supernatant, saliva, cerebrospinal fluid, semen, prostate fluid, tissue or tissue lysate.
  • Quantitative PCR was used to detect the expression of RBM46 gene in testicular cancer tissues.
  • Testicular cancer tissues and adjacent normal tissues were taken from the tumor hospital, placed in RNAlater reagent, and stored in a refrigerator at -80°C. First, extract total cellular RNA. Perform all operations at low temperature.
  • the A260/A280 value of 1.6 to 1.8 can ensure the purity of RNA.
  • the third step is to identify the expression level of Rbm46 gene by quantitative PCR. Add the reagents and concentrations shown in the table below to the PCR reaction tube and finally add the RNA sample.
  • reaction conditions were set in the PCR machine as follows: pre-denaturation at 95°C for 10s, denaturation at 95°C for 5s, annealing/extension at 60°C for 34s, a total of 40 cycles, with GAPDH as a relative quantitative internal control. 2 - ⁇ Ct method to calculate relative gene expression.
  • Electrophoresis Perform electrophoresis at 80V constant voltage, when bromophenol blue runs into the separation gel, switch to 120V constant voltage electrophoresis, and wait until the bromophenol blue dye runs to the bottom of the gel;
  • Clip "sandwich” black surface-sponge-3 filter paper-glue-PVDF membrane-3 filter paper-sponge-white surface, be careful not to leave bubbles.
  • RBM46 protein in testicular cancer tumors is shown in Figure 2.
  • the expression level of RBM46 protein in tumor tissues is significantly lower than that in adjacent tissues (p ⁇ 0.05).
  • the present disclosure relates to the application of a quantitative detection agent for Rbm46 mRNA or protein in the diagnosis and/or prognosis evaluation of testicular tumors, wherein the reduced expression of Rbm46 mRNA or protein is an indication of testicular tumors and/or poor prognosis of testicular tumors Indication.
  • Rbm46 mRNA or protein is highly expressed in normal cells and tissues of the testis, while it is significantly low expressed in testicular cancer cells.
  • the expression level is significantly correlated with clinicopathological parameters, and has important clinical application value.

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Abstract

La présente invention concerne une application de Rbm46 en tant que marqueur tumoral testiculaire. L'invention concerne une application d'un agent de détection quantitative pour l'ARNm ou la protéine Rbm46 dans la préparation d'un réactif ou d'un kit utilisé pour l'évaluation de diagnostic et/ou de pronostic d'une tumeur testiculaire; l'expression réduite de l'ARNm ou de la protéine Rbm46 étant une indication d'une tumeur testiculaire et/ou une indication d'un mauvais pronostic pour une tumeur testiculaire. La mesure de l'ARNm ou de la protéine Rbm46 peut être utilisée pour la détection ou le diagnostic de tumeurs testiculaires, ou l'évaluation et la surveillance du pronostic d'un patient, et présente une valeur d'application clinique importante.
PCT/CN2020/093005 2019-07-03 2020-05-28 Application de rbm46 en tant que marqueur tumoral testiculaire WO2021000673A1 (fr)

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CN201910595616.0 2019-07-03
CN201910595616.0A CN110195109A (zh) 2019-07-03 2019-07-03 Rbm46作为睾丸肿瘤标志物的应用

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017203526A1 (fr) * 2016-05-23 2017-11-30 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Méthodes de diagnostic du cancer à l'aide d'antigènes testiculaires du cancer
CN110195109A (zh) * 2019-07-03 2019-09-03 北京太东生物科技有限公司 Rbm46作为睾丸肿瘤标志物的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2524928A1 (fr) * 2011-05-18 2012-11-21 Atlas Antibodies AB RBM3 dans le cancer de la vessie

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017203526A1 (fr) * 2016-05-23 2017-11-30 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Méthodes de diagnostic du cancer à l'aide d'antigènes testiculaires du cancer
CN110195109A (zh) * 2019-07-03 2019-09-03 北京太东生物科技有限公司 Rbm46作为睾丸肿瘤标志物的应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
WAN, SHENGKAI ET AL: "National Academy of Clinical Biochemistry NACB Practice Guidelines: Tumor Markers in Testicular, Prostate, Colorectal, Breast and Ovarian Cancers", CHINESE JOURNAL OF CLINICAL LABORATORY SCIENCE, vol. 30, no. 2,, 28 February 2012 (2012-02-28) *

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