CN116536425A - Gene detection kit for tumor targeting and application thereof - Google Patents

Gene detection kit for tumor targeting and application thereof Download PDF

Info

Publication number
CN116536425A
CN116536425A CN202310535232.6A CN202310535232A CN116536425A CN 116536425 A CN116536425 A CN 116536425A CN 202310535232 A CN202310535232 A CN 202310535232A CN 116536425 A CN116536425 A CN 116536425A
Authority
CN
China
Prior art keywords
ccdc25a
lung cancer
canine lung
canine
monoclonal antibody
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.)
Withdrawn
Application number
CN202310535232.6A
Other languages
Chinese (zh)
Inventor
李丽伟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangzhou Gaohua Biotechnology Co ltd
Original Assignee
Guangzhou Gaohua Biotechnology Co ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Guangzhou Gaohua Biotechnology Co ltd filed Critical Guangzhou Gaohua Biotechnology Co ltd
Priority to CN202310535232.6A priority Critical patent/CN116536425A/en
Publication of CN116536425A publication Critical patent/CN116536425A/en
Withdrawn legal-status Critical Current

Links

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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • 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
    • C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Genetics & Genomics (AREA)
  • Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biotechnology (AREA)
  • Oncology (AREA)
  • Molecular Biology (AREA)
  • Hospice & Palliative Care (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention belongs to the technical field of biological diagnosis. The invention discloses application of CCDC25A in preparing a canine lung cancer targeted therapeutic drug and a diagnostic reagent. The invention also discloses a drug for targeted treatment of canine lung cancer, which comprises a bleomycin-coupled monoclonal antibody against CCDC25A. The invention also discloses a gene detection kit for evaluating the treatment effect of the canine lung cancer targeted drug, which comprises a reagent for detecting the expression level of CCDC25A. The invention provides application of a reagent for detecting CCDC25A in preparing a product for diagnosing canine lung cancer. The application comprises the kit for diagnosing the canine lung cancer and the preparation of the pharmaceutical composition for treating the canine lung cancer. The kit can be used as one of means for diagnosing the canine lung cancer, and is one of indexes for evaluating the effect of the canine lung cancer on CCDC25A targeted therapy. The pharmaceutical composition has a good inhibition effect on canine lung cancer and has important reference significance in canine lung cancer treatment.

Description

Gene detection kit for tumor targeting and application thereof
Technical Field
The invention belongs to the technical field of biological diagnosis, and particularly relates to a tumor targeting drug gene detection kit and application thereof.
Background
Primary lung tumors are rarely found in dogs, most are metastatic lung tumors, but the incidence of reported primary lung tumors may be increasing. This potential rise may reflect a number of factors, including increased exposure to environmental carcinogens and age-related canines, most of which are malignant. The benefit of annual geriatric canine physical examination (including chest X-ray examination) may be helpful in early diagnosis of lung cancer in asymptomatic animals.
Lung cancer, which may be primary (originating in the lung) or metastatic (originating from complications elsewhere in the body), is a serious disease that may be fatal if not discovered and treated early. Lung cancer rapidly spreads to other parts of the body, such as lymph nodes, bones, heart, liver, spleen and brain. If no treatment is found before the disease spreads to the lymph nodes, the prognosis is poor. Unfortunately, in all dogs with primary lung cancer, approximately one quarter has no symptoms other than tiredness and loss of appetite. Thus, early diagnosis and targeted therapy of canine lung cancer is particularly important. However, there is essentially no drug targeting canine lung cancer in the current market and a complete diagnostic method. Therefore, development of a target drug for canine lung cancer and detection matched with the target drug are urgently needed,
disclosure of Invention
The invention aims to solve the technical problems of screening a proper marker for diagnosing canine lung cancer; on the basis, a targeted therapeutic drug for canine lung cancer and a diagnostic reagent or kit of the targeted drug are developed.
Therefore, in one aspect, the invention provides an application of a canine lung cancer diagnosis marker, wherein the marker is CCDC25A, and the application of the CCDC25A in preparing a canine lung cancer targeted therapeutic drug and a diagnostic reagent.
Preferably, the nucleotide sequence of the CCDC25A is shown as SEQ ID NO. 1.
In yet another aspect, the invention also provides a medicament for targeted treatment of canine lung cancer, comprising a bleomycin conjugated monoclonal antibody directed against CCDC25A.
Preferably, the monoclonal antibody of the anti-CCDC 25A is a monoclonal antibody 4C10, wherein the nucleotide sequence of the heavy chain variable region of the monoclonal antibody 4C10 is shown as SEQ ID NO.4, and the nucleotide sequence of the light chain variable region of the monoclonal antibody 4C10 is shown as SEQ ID NO. 5.
In yet another aspect, the invention also provides a gene detection kit for evaluating the effect of a canine lung cancer targeted drug therapy, the kit comprising reagents for detecting CCDC25A expression levels.
Preferably, the reagent for detecting CCDC25A expression level according to the present invention comprises primers for specifically amplifying CCDC25A and reference beta-actin.
Preferably, the primers for specifically amplifying CCDC25A and reference beta-actin according to the present invention are as follows:
CCDC25A-F：5’-caagggtgcagtgaacttgc-3’；
CCDC25A-R：5’-agacggctgtacatctccct-3’；
β-actin-F：5’-gccagaaggactcctacgtg-3’；
β-actin-R：5’-agtccatcacgatgccagtg-3’。
the marker CCDC25A for diagnosing and treating the canine lung cancer is screened by bioinformatics and the existing molecular biological technical means. On the basis, the invention provides application of a reagent for detecting CCDC25A in preparing a product for diagnosing canine lung cancer. The application comprises the kit for diagnosing the canine lung cancer and the preparation of the pharmaceutical composition for treating the canine lung cancer. The kit can be used as one of means for diagnosing the canine lung cancer, and is one of indexes for evaluating the effect of the canine lung cancer on CCDC25A targeted therapy. The pharmaceutical composition has a good inhibition effect on canine lung cancer and has important reference significance in canine lung cancer treatment.
Drawings
FIG. 1 SDS-PAGE detection of CCDC25A protein.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art. Reagents and materials used in the following examples are commercially available unless otherwise specified.
"biomarker" and "marker" can be equivalently replaced, referring to a molecular indicator having a specific biological property, biochemical characteristic, or aspect, that can be used to determine the presence or absence of a particular disease or condition and/or the severity of a particular disease or condition. In the present invention, a "marker" refers to a parameter associated with one or more biomolecules (i.e., a "biomarker"), such as a naturally or synthetically produced nucleic acid (i.e., an individual gene, as well as coding and non-coding DNA and RNA). "marker" in the present invention also includes reference to a single parameter that may be calculated or otherwise obtained by considering expression data from two or more different markers.
Example 1: CCDC25A detection in canine lung cancer
The expression level of CCD25A in canine lung cancer was detected using conventional fluorescent PCR, and briefly described as follows:
1. sample collection is carried out by selecting a canine lung cancer case which is treated and diagnosed in an animal hospital in Guangzhou, and carrying out aseptic operation to leave partial cancer tissues and other normal tissues (about 2-5 cm away from the cancer tissues) and split charging the partial cancer tissues and other normal tissues into a freezing tube, and immediately storing the partial cancer tissues and the other normal tissues in liquid nitrogen. Of which 5 malignant tumor tissues and paracancerous tissues were selected for the study.
1.1 extraction of RNA from tissues
(1) The tissue samples were thoroughly ground with liquid nitrogen, 1ml of Trizol (Invitrogen) solution was added, mixed well, and left at room temperature for 5min for complete lysis;
(2) Adding 200 μl chloroform, shaking vigorously, mixing for 30s, allowing the water phase and the organic phase to fully contact, and standing at room temperature for 3-5min;
(3) At 4deg.C, 14000g was centrifuged for 15min to separate three layers, and RNA was transferred to another new RNase free EP tube in the upper aqueous phase;
(4) Adding equal volume of isopropanol, gently and fully mixing (reversing for 6-8 times), and standing at room temperature for 10min;
(5) Centrifuging 14000g for 10min at 4deg.C, collecting RNA precipitate, and removing supernatant;
(6) Washing with 75% ethanol twice (12000 g for 5 min), and air drying in a super clean bench; the sediment cannot be overdry or overdry, the sediment is not easy to dissolve when overdry and the ethanol remains when overdry;
(7) An appropriate amount of DEPC water (at least 15. Mu.l) was added depending on the amount of precipitate to dissolve the precipitate.
(8) RNA concentration was measured on the extracted RNA samples, and the ratio of OD260/OD280 was measured to control sample quality, and samples with a ratio of OD260/OD280 between 1.8 and 2.0 were selected for subsequent testing.
1.2 Primer design was performed using Primer 5.0 software based on CCDC25A (SEQ ID No. 1) sequence. Obtaining a plurality of pairs of specific primers, and finally respectively determining a group of optimal primers by comparison and screening, wherein beta-actin (shown as SEQ ID NO. 2) is used as an internal reference. The method comprises the following steps:
CCDC25A-F：5’-caagggtgcagtgaacttgc-3’；
CCDC25A-R：5’-agacggctgtacatctccct-3’；
β-actin-F：5’-gccagaaggactcctacgtg-3’；
β-actin-R：5’-agtccatcacgatgccagtg-3’。
1.3 reverse transcription reactions RNA reverse transcription reactions were performed using commercial kits. The specific operations are performed according to the specification. The reverse transcribed cDNA is stored at-80 deg.C for further use.
1.4 fluorescent quantitative PCR reaction the cDNA of the reverse transcription reaction product was taken for real-time fluorescent quantitative PCR reaction using a conventional dye method.
The reaction system was 20. Mu.l: mu.l SYBR Premix, 2. Mu.l cDNA template, 0.5. Mu.l upstream and downstream primer each and 7.0. Mu.l DEPC water. Experiments are carried out by using a fluorescent quantitative PCR instrument (such as Roche and the like), and the reaction setting conditions are as follows: pre-denaturation at 95℃for 30s, annealing at 58℃for 20s, elongation at 72℃for 30s,40 cycles.
The results show (table 1) that CCDC25A expression levels were significantly up-regulated in canine lung cancer (CT values were smaller compared to paracancerous tissues).
TABLE 1 detection results of CCDC25A in canine lung cancer
Project CCDC25ACT value Beta-actin CT value
Cancer tissue 15.45 11.36
Tissue beside cancer 25.98 11.45
Example 2: treatment of canine lung cancer against CCDC25A target
1. Preparation and testing of CCDC25A monoclonal antibodies
Preparation of 1.1CCDC25A protein
The CCDC25A protein was prepared using a conventional eukaryotic expression system, the steps of which are briefly described below: the sequence of SEQ ID No.1 was codon optimized, and the nucleotide sequence of the codon optimized CCDC25A protein (shown as SEQ ID No. 3) was used as a template to construct pEE12.4-CCDC25A-6His expression plasmid by ligating with EcoRI and XhoI double cleavage sites, respectively. The recombinant plasmid identified as positive was sequenced. Transiently transfecting the CHO cells with the identified correct plasmid, harvesting cell culture supernatant after 72 hours, and purifying the secreted and expressed CCDC25A protein by using a nickel column; the BCA kit was used to determine concentration and purity by SDS-PAGE. The SDS-PAGE purity of CCDC25A protein (FIG. 1) > 90% was determined to be 0.89mg/ml. Quantitatively packaging the protein, and storing at-80deg.C.
1.2 preparation of anti-CCDC 25A protein monoclonal antibody
Immunizing a BALB/c mouse with 6-8 weeks old by using the CCDC25A protein prepared by 1.1, carrying out isovolumetric emulsification on the CCDC25A protein and Freund's complete adjuvant for the first time, and inoculating 100 mug protein/mouse in the abdominal cavity; after 7 days, the CCDC25A protein is emulsified with Freund's incomplete adjuvant in equal volume, and the mice are immunized by the second intraperitoneal inoculation route, wherein 100 mug protein/mouse; the third mouse peritoneal route directly immunized with CCDC25A protein after 7 days, 100 μg/mouse; on day 3 after immunization, taking mouse spleen cells and mouse myeloma cells SP2/0 for fusion, and culturing in HAT selective medium; after 10 days, CCDC25A protein is taken as a coating antigen, cell supernatant is detected by indirect ELISA, positive hybridoma cells are screened, and a better hybridoma cell strain is screened from the positive hybridoma cell strain, and is named as hybridoma cell 4C10 strain.
Taking 8-10 week old Balb/C mice, injecting pristane intraperitoneally, each 0.5ml, and injecting hybrid 4C10 strain of hybridoma cells 1X 10 into each mice 7 days later 6 After 7-10 days, the ascites of the mice are extracted, and the supernatant is collected by centrifugation at 1200r/min for 10 minutes at 2-8 ℃. The monoclonal antibodies were purified using a ProteinG affinity column, and the purified monoclonal antibodies (designated monoclonal antibody 4C 10) were individually packaged into 0.5 ml/tube and stored at-80℃for further use.
Analysis of the sequence of monoclonal antibody 4C10 revealed that the sequences encoding the heavy and light chain variable regions of monoclonal antibody 4C10 are shown in SEQ ID NO.4 and SEQ ID NO.5, respectively.
2. Coupling of monoclonal antibodies to bleomycin
SPDP (3- (2-pyridine dimercapto) propionic acid N-hydroxysuccinimide ester) is used as a connecting agent, the molar ratio of the monoclonal antibody 4C10 to the SPDP is 1:10, the molar ratio of bleomycin to the SPDP is 1:1, the sulfhydrylation monoclonal antibody 4C10 and the sulfhydrylation bleomycin are mixed, DTT (dithiothreitol) is added, the conjugate part is collected through SephadexG 25 column chromatography, the protein content is detected by a BCA method, and the protein is stored at the temperature of 80 ℃ below zero for standby. The conjugate is detected by ultraviolet-visible spectrophotometry and mass spectrometry, and the molar ratio of the antibody to bleomycin in the conjugate is 1:3-5.
3. Treatment targeting therapy and gene detection evaluation of canine lung cancer
The prepared monoclonal antibody targeting medicine coupled with bleomycin is used for treating dogs with clinically diagnosed lung cancer, the dogs are treated by intravenous injection (injected according to the amount of 1 mg/kg), serum is collected weekly after treatment to detect the expression amount of CCDC25A, and meanwhile, the traditional chemotherapy mode is used for parallel treatment and detection, so that the feasibility and the effectiveness of the treatment scheme are comprehensively evaluated, and the effect of the targeting treatment is evaluated in real time.
The treatment results show that the treatment effect of the targeted drug provided by the invention is better than that of the traditional chemotherapy method, and the treatment effect is mainly characterized by smaller side effects and longer life cycle (generally more than half a year or more) of dogs in the treatment process.
Meanwhile, the effect of the targeted therapy is evaluated by using the method of the embodiment 1, and the expression quantity (CT value is 25 or more) of CCDC25A indicates that the therapeutic effect is better, and the basic survival state of dogs is better; when the CT value is less than or equal to 20, intravenous administration (targeted drug, 1 mg/kg); when the CT value is more than 20 and less than 25, the living state of dogs is concerned, the detection is enhanced, and the medicine is timely administered.
Of course, those skilled in the art can apply the targeting drug of the present invention while using the conventional therapeutic scheme or other schemes on the basis of the present invention, and shall also fall within the technical scheme of the present invention.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims (7)

1. The application of the canine lung cancer diagnosis marker is characterized in that the marker is CCDC25A, and the CCDC25A is applied to preparing a canine lung cancer targeted therapeutic drug and a diagnostic reagent.
2. The use according to claim 1, wherein the nucleotide sequence of CCDC25A is shown in SEQ ID No. 1.
3. A medicament for targeted treatment of canine lung cancer, wherein the medicament comprises a bleomycin conjugated monoclonal antibody directed against CCDC25A.
4. The agent of claim 3, wherein the monoclonal antibody against CCDC25A is monoclonal antibody 4C10, wherein the nucleotide sequence encoding the heavy chain variable region of monoclonal antibody 4C10 is shown in SEQ ID No.4, and the nucleotide sequence encoding the light chain variable region of monoclonal antibody 4C10 is shown in SEQ ID No. 5.
5. A gene detection kit for evaluating the effect of a canine lung cancer targeted drug therapy, which is characterized by comprising a reagent for detecting the expression level of CCDC25A.
6. The kit of claim 5, wherein the reagent for detecting CCDC25A expression level comprises primers for specifically amplifying CCDC25A and reference β -actin.
7. The kit according to claim 6, wherein the primers for specifically amplifying CCDC25A and reference β -actin are as follows:
CCDC25A-F：5’-caagggtgcagtgaacttgc-3’；
CCDC25A-R：5’-agacggctgtacatctccct-3’；
β-actin-F：5’-gccagaaggactcctacgtg-3’；
β-actin-R：5’-agtccatcacgatgccagtg-3’。
CN202310535232.6A 2023-05-12 2023-05-12 Gene detection kit for tumor targeting and application thereof Withdrawn CN116536425A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310535232.6A CN116536425A (en) 2023-05-12 2023-05-12 Gene detection kit for tumor targeting and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310535232.6A CN116536425A (en) 2023-05-12 2023-05-12 Gene detection kit for tumor targeting and application thereof

Publications (1)

Publication Number Publication Date
CN116536425A true CN116536425A (en) 2023-08-04

Family

ID=87457462

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310535232.6A Withdrawn CN116536425A (en) 2023-05-12 2023-05-12 Gene detection kit for tumor targeting and application thereof

Country Status (1)

Country Link
CN (1) CN116536425A (en)

Similar Documents

Publication Publication Date Title
WO2012017208A1 (en) Methods and compounds for the diagnosis and treatment of
KR102210333B1 (en) Composition for diagnosing cancer
JP2014501918A (en) AGTR1 as a marker for bevacizumab combination therapy
JP2022519649A (en) How to diagnose and treat cancer
CN107488737B (en) Liquid biopsy kit for detecting peripheral blood TCR variable region coding gene and application thereof
JP6908530B2 (en) Screening for muscle atrophy-related protein molecular marker Dkk-3 and its use
CN112626207A (en) Gene combination for distinguishing non-invasive and invasive non-functional pituitary adenomas
CN110251669B (en) CXCL16 protein and application of monoclonal antibody thereof in preparation of medicines for preventing and/or treating intestinal injury diseases
CN106701902B (en) Application of FOXR2 gene and expression product in diagnosis and treatment of liver cancer
CN108559772B (en) MARCH1 gene as diagnosis and treatment marker for rheumatoid arthritis and osteoarthritis
CN108531607B (en) Diagnosis marker-C16orf74 the gene of clear cell carcinoma of kidney
CN116536425A (en) Gene detection kit for tumor targeting and application thereof
US10370417B2 (en) Methods for the diagnosis of cancer through detection of human endogenous retrovirus WL (HERV-WL) envelope expression
CN106947818B (en) Molecular marker for diagnosis and treatment of colon adenocarcinoma
CN105753960B (en) A kind of tumor markers and its application
CN108642168B (en) Diagnosis and treatment of rheumatoid arthritis and osteoarthritis based on SPAST gene
CN106947819B (en) Marker for diagnosis and treatment of colon adenocarcinoma
CN106834496B (en) PNLIPRP3 gene and application of expression product thereof in diagnosis and treatment of tongue squamous cell carcinoma
WO2007043531A1 (en) Genetic marker for use in diagnosis of sensitivity to anti-tumor agent or concurrent chemoradiation therapy, and use thereof
LU503219B1 (en) Lncrna bdnf-as and applications thereof as marker and therapeutic target
CN108048568B (en) Application of PLCD4 gene as gastric adenocarcinoma metastasis diagnosis marker
WO2019056199A1 (en) Ovarian cancer biomarker and use thereof
WO2008041767A1 (en) Gene/protein marker for prediction or diagnosis of pharmacological efficacy of aurora a inhibitor
CN110499365B (en) Application of AGAP9 in preparation of product for diagnosing osteoarthritis
KR20190048327A (en) Biomarker for diagnosis of anticancer drug resistance of gastric cancer and use thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20230804