CN105886660A - Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker - Google Patents
Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker Download PDFInfo
- Publication number
- CN105886660A CN105886660A CN201610494676.XA CN201610494676A CN105886660A CN 105886660 A CN105886660 A CN 105886660A CN 201610494676 A CN201610494676 A CN 201610494676A CN 105886660 A CN105886660 A CN 105886660A
- Authority
- CN
- China
- Prior art keywords
- uhrf1
- gene
- albumen
- expression
- carcinoma
- 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.)
- Pending
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
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- 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
- G01N33/57442—Specifically defined cancers of the uterus and endometrial
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Pathology (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Hospice & Palliative Care (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Oncology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Reproductive Health (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a gene marker UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1). UHRF1 can be used for judging whether subjects have the risk of suffering from endometrial cancers or judging whether the subjects suffer from endometrial cancers. Besides, UHRF1 can be also used for preparing medicines for treating endometrial cancer. A new diagnostic method is provided for clinically diagnosing endometrial cancers at a molecular level, and meanwhile, a new medicine target is provided for gene treatment of endometrial cancers.
Description
Technical field
The present invention relates to cancer diagnosis, treat, predict prognosis field, more particularly it relates to detection
UHRF1 is abnormal is the cancer diagnosis of means, prediction method of prognosis；And suppression UHRF1 gene or protein
Cancer therapeutic agent.
Background technology
Carcinoma of endometrium is one of big malignant tumor of female genital tract three, pernicious in American-European its sickness rate Yi Zhan gynecological
First of tumor.In recent years, along with the developing rapidly of China's economy, people's living habit and dietary structure
Change, the factor such as non-normal hormone replacement therapy and gonadal hormone abuse, the incidence rate of carcinoma of endometrium substantially on
Rise, and tend to rejuvenation, become the reproductive tract malignant disease of serious threat China women's health.
Carcinoma of endometrium accounts for the 7% of female malignant sum, annual neopathy number of cases and death number respectively
Account for the 3.9% and 1.7% of female malignant.Due to interzone economy and the difference of medical service provider system level,
Different to the level of endometrial carcinomas early diagnosis and therapy, prognosis is the most significantly different.Although in developing country uterus
The sickness rate of endometrial carcinomas is less than developed country, but its mortality rate/sickness rate is higher than developed country.Developing country
Mortality rate/sickness rate be 34% (21 000,/62 000), within 5 years, survival rate is 67%, and the death of developed country
Rate/sickness rate is 21% (29 0,00/,136 000), and within 5 years, survival rate is 82%.
The high risk factor of carcinoma of endometrium include obesity, hypertension, diabetes, menopause postpone, infertile and long-term
Simple estrogen stimulation etc..The age occurred frequently of endometrial carcinomas is 50～60 years old, is main with abnormal vaginal bleeding
Performance, during ID, 72% was I phase, and 12% was II phase, and 13% was III phase, and 3% was IV phase, as sent out in early days
Existing early treatment, prognosis is preferable.
Summary of the invention
An object of the present invention is that providing a kind of is examined by detection UHRF1 gene or protein expression difference
The method of disconnected carcinoma of endometrium.
The two of the purpose of the present invention are that providing a kind of comes pre-by detection UHRF1 gene or protein expression difference
The method surveying carcinoma of endometrium prognosis.
The three of the purpose of the present invention are that providing a kind of is controlled by suppression UHRF1 gene or UHRF1 albumen
The method treating carcinoma of endometrium.
A kind of method that the four of the purpose of the present invention are to provide medicine for screening treatment carcinoma of endometrium.
The five of the purpose of the present invention are to provide a kind of medicine for treating carcinoma of endometrium.
To achieve these goals, present invention employs following technical scheme:
The product that the invention provides detection UHRF1 gene or UHRF1 albumen is examined in preparation carcinoma of endometrium
Purposes in disconnected instrument.
Present invention also offers the product of detection UHRF1 gene or UHRF1 albumen at preparation prediction intrauterine
Purposes in film cancer prognostic tool.
Further, the product of described detection UHRF1 gene or UHRF1 albumen includes detecting UHRF1 gene
Or the product of the expression of UHRF1 albumen.Described product includes can be in conjunction with the nucleic acid of UHRF1 gene
Or can be in conjunction with the material (such as antibody) of UHRF1 albumen.Described nucleic acid can detect UHRF1 gene
Expression；Described material can detect the expression of UHRF1 albumen.
The product of the detection UHRF1 gene of the present invention can play it based on the known method using nucleic acid molecules
Function: as PCR, as Southern hybridization, Northern hybridization, dot blot, fluorescence in situ hybridization (FISH),
DNA microarray, ASO method, high-flux sequence platform etc..Use this product can qualitatively, quantitatively,
Or semi-quantitatively implement to analyze.
The nucleic acid being included in the said goods can be obtained by chemosynthesis, or by preparing from biomaterial
Containing expectation nucleic acid gene, then use be designed for amplification expectation nucleic acid primer amplification it obtain.
Further, described PCR method is known method, such as, and ARMS (Amplification Refractory
Mutation System, amplification do not answer abruptly-changing system) method, RT-PCR (reverse transcriptase-PCR) method, nesting
PCR method etc..The nucleic acid of amplification can be by using dot blotting hybridization method, Surface Plasmon Resonance (SPR
Method), PCR-RFLP method, B by means of in situ RT PCR, PCR-SSO (sequence specific oligonucleotide) method,
PCR-SSP method, AMPFLP (amplifiable fragment length polymorphism) method, MVR-PCR method and
PCR-SSCP (single strand conformation polymorphism) method detects.
Nucleic acid recited above includes the primer expanding UHRF1 gene, and the primer that product includes can pass through
Being prepared by chemosynthesis, the method that be those skilled in the art will know that by use is come suitably with reference to Given information
Ground design, and prepared by chemosynthesis.
In specific embodiments of the present invention, described nucleic acid is the amplimer used in QPCR experiment, institute
State shown in sequence such as SEQ ID NO.1 (forward sequence) and SEQ ID NO.2 (reverse sequence) of primer.
Nucleic acid recited above may also include probe, and described probe can be prepared by chemosynthesis, by making
Appropriately design with reference to Given information by the method that those skilled in the art will know that, and prepared by chemosynthesis,
Or the gene containing expectation nucleotide sequence from biomaterial preparation can be passed through, and use is designed for the amplification phase
Hope nucleotide sequence primer amplification it prepare.
The product of the detection UHRF1 albumen of the present invention can play its function based on the known method using antibody:
For example, it is possible to include ELISA, radioimmunoassay, immunohistochemical method, western blot etc..
The product of the detection UHRF1 albumen of the present invention include the antibody of specific binding UHRF1 albumen or its
Fragment.Antibody or its fragment of any structure, size, immunoglobulin class, origin etc. can be used, only
It is wanted to combine target protein.Antibody or its fragment that the detection product of the present invention includes can be monoclonals
Or polyclonal.Antibody fragment refers to an antibody part (Partial Fragment) retaining antibody to the combination activity of antigen
Or the peptide containing an antibody part.Antibody fragment can include F (ab ')2, Fab ', Fab, scFv (scFv),
The Fv (dsFv) of disulphide bonding or its polymer, dimerization V district (double antibody) or containing CDR
Peptide.The product of the detection UHRF1 albumen of the present invention can include the amino of encoding antibody or Encoding Antibody Fragment
The nucleic acid of the separation of acid sequence, comprises the carrier of this nucleic acid, and carries the cell of this carrier.
Antibody can be by well known to a person skilled in the art that method obtains.Such as, preparation retains whole or portion
The polypeptide dividing target protein or the mammalian cell expression vector integrating their polynucleotide of coding are as anti-
Former.Use after antigen-immunized animal, from through the animal adaptive immune cell of immunity fused bone myeloma cells with
Obtain hybridoma.Then antibody is collected from Hybridoma culture.Finally can be used as antigen by use
UHRF1 albumen or its part antibody to obtaining are implemented antigenic specificity purification and are obtained for UHRF1 egg
White monoclonal antibody.Polyclonal antibody can be prepared as follows: with antigen-immunized animal same as above, from
Animal through immunity collects blood sample, isolates serum, then use above-mentioned antigen to serum from blood
Implement antigenic specificity purification.Can be by the antibody obtained with ferment treatment or the sequence of the antibody obtained by use
Column information obtains antibody fragment.
The combination of label and antibody or its fragment can be implemented by method as commonly known in the art.Such as,
Can fluorescent marker protein or peptide as follows: clean protein or peptide with phosphate buffer, add with DMSO,
Buffer agent, etc. preparation dyestuff, then mixed solution, then at room temperature place 10 minutes.It addition, labelling can
The labelling kit of commodity in use, such as biotin labeling reagent box, as biotin labeling reagent box-NH2,
Biotin labeling reagent box-SH (DojindoLaboratories)；Alkali phosphatase enzyme mark test kit such as alkalescence phosphorus
Acid enzyme labelling test kit-NH2, alkali phosphatase enzyme mark test kit-SH (Dojindo Laboratories)；Peroxide
Compound enzyme labelling test kit such as peroxidase labelling test kit-NH2, peroxidase labelling test kit
-NH2(Dojindo Laboratories)；Phycobniliprotein labelling kit such as phycobniliprotein labelling kit
-NH2, phycobniliprotein labelling kit-SH, B-phycoerythrin labelling kit-NH2, B-phycoerythrin
Labelling kit-SH, R-PE labelling kit-NH2, R-PE labelling kit
SH(DojindoLaboratories)；Fluorescent labeling reagent box such as fluorescein labelling kit-NH2, HiLyte
Fluor (TM) 555 labelling kit-NH2, HiLyte Fluor (TM) 647 labelling kit-NH2 (Dojindo
Laboratories)；And DyLight 547 and DyLight647 (Techno Chemical Corp.), Zenon (TM),
Alexa Fluor (TM) antibody labeling test kit, Qdot (TM) antibody labeling test kit (Invitrogen
And EZ-label Protein Labeling Kit (Funakoshi Corporation) Corporation).In order to correctly
Labelling, it is possible to use suitable instrument detects the antibody through labelling or its fragment.
Sample as the detection product according to the present invention, it is possible to use the tissue such as obtained from biopsy experimenter
Sample or fluid.Sample is not particularly limited, as long as it is suitable to the mensuration of the present invention；Such as, it can include
Tissue, blood, blood plasma, serum, lymph fluid, urine, serous cavity liquid, spinal fluid, synovial fluid, aqueous humor, tear
Liquid, saliva or its fraction or treated material.
In specific embodiments of the present invention, described sample is from the tissue of experimenter.
In the present invention, " prognosis " refer to that cancer patient is by the suppression such as surgical procedure or alleviate tumor growth
After process or result.In this manual, prognosis can be to be suppressed by surgical procedure or alleviate tumor growth
Life state when latter 1,2,3,4,5,6,7,8,9,10,15,20 years or more long.Prognosis can
With by checking that the gene of biomarker i.e. UHRF1 albumen or coding UHRF1 albumen is predicted.Prognosis
Prediction can be performed such that according to biomarker with or without, or be raised and lowered, determine that patient's is pre-
After be good or bad, or determine the probability of good prognosis or poor prognosis.
In the present invention, " prognosis bona " refers to suppressed or alleviation tumor life for patient by surgical procedure etc.
After length, patient (such as 3,5,6,7,8,9,10,15,20 years or longer) over a long time not danger
Anxious situation.Or, good prognosis can mean to survive in the most long-time, without transfer, without recurrence or without again
Send out.Such as, prognosis bona can mean at least 3 years or survival in especially at least 5 years, preferably without transfer
Or recurrence.The most preferred state of prognosis bona is the long-term survival without disease.As used herein, " pre-
Rear good " can also include any such state, wherein it appeared that disease such as transfer, but pernicious low and
Do not severely impact survival ability.
In the present invention, " prognosis mala " refers to that patient is by the suppression such as surgical procedure or alleviation tumor growth
After short-term (such as 1,2,3,4,5 years or shorter) in occur fatal condition.Or, poor prognosis is
Refer to death in such short-term, shift, recur or send out again.Such as, poor prognosis can mean at least 3
Year or Preventive or death in especially at least 5 years.
Prediction prognosis refers to predict process or the result of status of patient, and being not meant to can be with the accuracy of 100%
The process of prediction status of patient or result.Prediction prognosis refers to whether the probability determining some process or result increases
Add, and be not meant to be compared by situation about not occurring with some process or result determine generation some process
Or the probability of result.For the present invention, UHRF1 gene or the level of UHRF1 albumen in the present invention
In the patient raised, compared with the patient not showing this feature, more likely observe particular procedure or result.
Further, the product of described detection UHRF1 gene or UHRF1 albumen can be detection UHRF1 base
Because of or the reagent of UHRF1 albumen, can also be to comprise the test kit of described reagent, chip, reagent paper etc., also
It can be the high-flux sequence platform using described reagent.
Present invention also offers the instrument of a kind of diagnosis of endometrial carcinoma, described instrument can detect UHRF1 base
Cause or the expression of UHRF1 albumen.Described instrument include can in conjunction with the nucleic acid of UHRF1 gene or
Can be in conjunction with the material (such as antibody) of UHRF1 albumen.Described nucleic acid can detect the table of UHRF1 gene
Reach level；Described material can detect the expression of UHRF1 albumen.
Further, the character of described nucleic acid and described material is with noted earlier.
Further, the instrument of described diagnosis of endometrial carcinoma include but not limited to chip, test kit, reagent paper or
High-flux sequence platform；High-flux sequence platform is the instrument of a kind of special diagnosis of endometrial carcinoma, along with height
The development of flux sequencing technologies, will become the structure of the gene expression profile of a people and work the most easily.Logical
Cross contrast Disease and the gene expression profile of normal population, easily analyze exception and the disease phase of which gene
Close.Therefore, in high-flux sequence, know that the exception of UHRF1 gene is relevant to carcinoma of endometrium fall within
The purposes of UHRF1 gene, equally within protection scope of the present invention.
Present invention also offers a kind of instrument predicting carcinoma of endometrium prognosis, described prediction carcinoma of endometrium prognosis
Instrument includes can be in conjunction with the nucleic acid of UHRF1 gene or can in conjunction with the material of UHRF1 albumen (such as
Antibody).Described nucleic acid can detect the mRNA level in-site of UHRF1 gene；Described material can detect UHRF1
The expression of albumen.
Further, the character of described nucleic acid and described material is with noted earlier.
Further, the instrument of described prediction carcinoma of endometrium prognosis include but not limited to chip, test kit, reagent paper,
Or high-flux sequence platform；High-flux sequence platform is the instrument of a kind of special diagnosis of endometrial carcinoma, along with
The development of high throughput sequencing technologies, will become the structure of the gene expression profile of a people and work the most easily.
By contrast Disease and the gene expression profile of normal population, easily analyze exception and the disease of which gene
Relevant.Therefore, in high-flux sequence, know that the exception of UHRF1 gene is relevant to carcinoma of endometrium fall within
The purposes of UHRF1 gene, equally within protection scope of the present invention.
The ammonia that the anti-UHRF1 antibody used in detection product, the diagnostic tool of the present invention or its fragment are identified
The number of base acid is not particularly limited, as long as antibody can be in conjunction with UHRF1.When antibody is as curative
During thing, preferably it is capable of identify that aminoacid as much as possible, as long as it can suppress UHRF1 function.Anti-
The amino acid whose number of body or its fragment identification is at least one, more preferably at least three.The immune globulin of antibody
White classification is unrestricted, can be IgG, IgM, IgA, IgE, IgD or IgY.
Present invention also offers a kind of diagnosis of endometrial carcinoma or the method for prediction carcinoma of endometrium prognosis, described side
Method comprises the steps:
(1) sample of experimenter is obtained；
(2) UHRF1 gene or the expression of albumen in detection Samples subjects；
(3) the UHRF1 gene recorded or the expression of albumen have been associated with the whether ill of experimenter
Come.
(4) compared with the control, the expression of UHRF1 gene or albumen raises, then this experimenter is diagnosed
For carcinoma of endometrium, or this experimenter is confirmed as prognosis mala.
Present invention also offers the Therapeutic Method of a kind of carcinoma of endometrium, described method includes suppressing UHRF1 base
Cause or UHRF1 albumen.
Further, described method includes the expression suppressing UHRF1 gene, or the table of suppression UHRF1 albumen
Reach or suppress the activity of UHRF1 albumen.
Present invention also offers the screening technique of a kind of cancer drug, can be by cancerous cell being added test medicine
Measure certain period after thing or after cancer model animal is used testing drug UHRF1 gene or
The expression of UHRF1 albumen measures cancer drug and improves the effect of cancer prognosis.More specifically, when
The expression of UHRF1 gene or UHRF1 albumen when adding or reduce after using testing drug or
When recovering normal level, this medicine optional is as the medicine improving cancer prognosis.
Present invention also offers a kind of containing UHRF1 gene or the medicine of the inhibitor of UHRF1 albumen.
Present invention also offers the application in the medicine of preparation treatment carcinoma of endometrium of the above-mentioned inhibitor.
The UHRF1 gene of the present invention or the inhibitor of UHRF1 albumen are unrestricted, as long as can suppress
UHRF1 or relate to the expression of material or the activity of UHRF1 upstream or downstream pathway, and treatment cancer is had
The medicine of effect.
Further, described inhibitor includes that antisensenucleic acids, dsRNA, ribozyme, fit, UHRF1 combine egg
White tiles section or antibody or its fragment.
" antisensenucleic acids " refers to containing the nucleic acid with the sequence of the mRNA complementation of coding UHRF1.Antisensenucleic acids
Can by DNA, RNA or the two form.Antisensenucleic acids need not the mRNA 100% with target UHRF1
Complementary.Antisensenucleic acids can contain Non-complementary bases, as long as it can specific hybrid under strict conditions.
When antisensenucleic acids is introduced cell, it combine target polynucleotide and suppression is transcribed, RNA processing, translation or
Stability.In addition to antisense polynucleotides, antisensenucleic acids also includes polynucleotide analogies, and it contains through repairing
The main chain of decorations and 3 ' and 5 ' end portion.Such antisensenucleic acids can come according to UHRF1 sequence information
Appropriate design use well known to a person skilled in the art that method generates.
" dsRNA " refers to, containing duplex-RNA constructs, carry out inhibition of gene expression by RNA interference (RNAi)
RNA, including siRNA (short interfering rna) and shRNA (short hairpin RNA).DsRNA need not
With the homology that target-gene sequence has 100%, as long as it can suppress expression of target gene.For stabilisation
Or other purpose, a part of DNA of dsRNA can be substituted.Preferably, siRNA is 21-23
The double-stranded RNA of individual base.SiRNA can be by well known to a person skilled in the art prepared by method, such as
By chemosynthesis or as the analog naturally occurring RNA.ShRNA is to have hair clip corner (hairpin
Turn) Short interfering RNA of structure.ShRNA can by well known to a person skilled in the art prepared by method,
Such as by chemosynthesis or by the DNA of coding shRNA is introduced cell expressible dna.
" ribozyme " refers to the RNA with catalysis activity, and it can cut, pastes, insert and transfer RNA.
The structure of ribozyme can include tup, hair clip etc..
" fit " refers to combine the nucleic acid of something such as protein.Fit can be RNA or DNA.Nucleic acid
Form can be double-strand or strand.Fit infinite in length system, as long as it can specific binding target molecule be
Can, can by such as 10 to 200 nucleotide, preferably 10 to 100 nucleotide, more preferably 15 to
80 nucleotide, even more preferably 15 to 50 nucleotide compositions.Fit can use art technology
Known to personnel, method selects.It is for instance possible to use SELEX (is enriched with the part carried out by exponential form
Phyletic evolution).
" the protein-bonded fragment of UHRF1 " refers to combine UHRF1 and suppression UHRF1 implements original function
The fragment of protein.
The medicine of the present invention can be administered alone as medicine or use together with other medicines.Can be with the present invention
The other medicines used together of medicine unrestricted, as long as it does not damage the therapeutic of the present invention or preventative medicine
The effect of thing, it is preferred that the medicine for treatment or prophylaxis of cancer can include such as alkylating agent, all
As ifosfamide, cyclophosphamide, dacarbazine, temozolomide, nimustine, busulfan, procarbazine,
Melphalan and Ranimustine；Antimetabolite, such as enocitabine, capecitabine, carmofur, cladribine,
Gemcitabine, cytosine arabinoside, cytosine arabinoside octadecyl phosphate (cytarabine ocfosfate), ftorafur,
UFT, ftorafur gimeracil oteracil potassium, doxifluridine, hydroxyurea, fluorouracil,
Fludarabine, pemetrexed, pentostatin, mercaptopurine and methotrexate；Plant alkaloid, such as Yi Li
For health, etoposide, sobuzoxane, docetaxel, nogitecan, Pa Litasai, vinorelbine, Changchun
Pungent and the vinblastine in ground；Antitumor antibiotic, such as actinomycin D, aclarubicin, amrubicin, Yi Da ratio
Star, epirubicin, zinostatin stimalamer, daunorubicin, doxorubicin, pirarubicin, rich come mould
Element, peplomycin, ametycin and mitoxantrone；Medicine based on platinum, such as oxaliplatin, carboplatin,
Cisplatin and nedaplatin；Hormonal medicaments, such as Anastrozole, exemestane, estramustine, ethinylestradiol, chlorine
Ground progesterone, goserelin, tamoxifen, dexamethasone, toremifene, bicalutamide, flutamide, bold and vigorous Buddhist nun
Song Long, fostestrol, mitotane, methyltestosterone, medroxyprogesterone, mepitiostane, leuprorelin and letrozole；Raw
Thing reaction dressing agent, such as interferon-ALPHA, interferon beta, interferon gamma, interleukin, ubenimex, dry BCG,
And lentinan；And molecular targeted agents, such as imatinib (imatinib), gefitinib (gefitinib), Ji
Nurse monoclonal antibody, ozogamicin, Tamibarotene, trastuzumab, tretinoin, bortezomib (bortezomib),
With Rituximab etc..
The medicine of the present invention can be prepared as various dosage form as required.Include but not limited to, percutaneous, mucosa, nose,
Buccal, Sublingual or the tablet of per os use, solution, granule, patch, unguentum, capsule, aerosol
Or suppository.
The route of administration of the medicine of the present invention is unrestricted, as long as it can play desired therapeutic effect or pre-preventive effect
Fruit, includes but not limited to intravenous, intraperitoneal, ophthalmic, intra-arterial, in lung, is administered orally, in vesicle,
Intramuscular, tracheal strips, subcutaneous, by skin, by pleura, local, suck, by mucosa, skin
Skin, the intestines and stomach, intraarticular, in ventricle, rectum, vagina, in skull, in urethra, in liver, in tumor.At certain
In the case of Xie, can systematically be administered.It is to be administered partly in some cases.
The dosage of the medicine of the present invention is unrestricted, as long as obtaining desired therapeutic effect or preventive effect,
Appropriate determination can be carried out according to symptom, sex, age etc..The medicine of the present invention or the agent of prophylactic agent
Amount can use such as therapeutic effect or preventive effect to disease to determine as index.
In the context of the present invention, " diagnosis of endometrial carcinoma " both includes judging that experimenter has suffered from
Carcinoma of endometrium, also include judging whether experimenter exists the risk suffering from carcinoma of endometrium.
" treatment " used herein contains treatment phase in the mammal such as mankind suffering from relevant disease or disease
The disease closed or morbid state, and include:
(1) prevention disease or morbid state occur in mammal, especially susceptible in institute when this mammal
State morbid state, but be not yet diagnosed when suffering from this morbid state；
(2) suppression disease or morbid state, i.e. stop it to occur；Or
(3) disease or morbid state are alleviated, even if disease or morbid state disappear.
Term " is treated " and is usually directed to treat the mankind or animal (such as, applied by veterinary), wherein can reach certain
Some intended therapeutic effect, such as, the development (including reducing development speed, making development stop) of suppression disease,
Improve disease and cure disease.Also include the treatment as preventive measure (such as prevention).To the most not developing into
Disease but have develops into the purposes of the dangerous patient of this disease, is also included within during term " treats ".
Advantages of the present invention and beneficial effect:
The molecular marker being found that a kind of diagnosis of endometrial carcinoma of the present invention, uses this molecular marker permissible
The early stage that Endometrial Carcinomas occurs can be used as judging, it is provided that the survival rate of patient.
It addition, by the prognosis predicting patient, the present invention can provide significant information to determine to control for patient
Treat scheme policies.
The medicine of the inhibitor including UHRF1 gene or albumen of the present invention can be used as new endometrium
The medicine of cancer.
Accompanying drawing explanation
Fig. 1 shows and utilizes in QPCR detection UHRF1 gene endometrial tissues and normal endometrial tissue
Expression；
Fig. 2 shows and utilizes Western blot detection UHRF1 albumen endometrial tissues and normal endometrium group
Expression in knitting；
Fig. 3 show utilize QPCR detect the siRNA jamming effectiveness to UHRF1 gene；
Fig. 4 shows the impact that endometrial carcinoma cell is bred by suppression UHRF1 protein function.
Specific embodiment
The present invention is further detailed explanation with embodiment below in conjunction with the accompanying drawings.Following example are only used for
The bright present invention rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in embodiment is logical
Often according to normal condition, such as Sambrook et al., molecular cloning: laboratory manual (New York:Cold
Spring HarborLaboratory Press, 1989) condition described in, or according to the bar proposed by manufacturer
Part.
The differential expression of embodiment 1 UHRF1 gene
1, sample collection:
Endometrial sample: collect endometrial carcinoma, the equal underwent operative of all patients is treated, operation
Paraffin specimen 10 example.All patients all make a definite diagnosis through pathologic finding suffers from carcinoma of endometrium.Other enter set condition
Any treatment was not accepted: do not merge other malignant tumor before being admitted to hospital for: all patients；Do not merge other hormones
Diseases related；Complete clinical data.
10 patients with endometrial cancer, fall ill 58 years old mean age.Patient's main clinical manifestation is that vagina is not advised
The most hemorrhage, lower abdominal pain, menoxenia, neoplasm etc., also have some patients non-evident sympton to look in health
Body finds.Carcinoma of endometrium specimen is diagnosed as carcinoma of endometrium through HE stained, tectology.
Normal endometrial tissue sample: 10 example normal endometrium operation paraffin specimens.The disease of samples sources
People's illnesses includes: hysteromyoma, uterine prolapse, wing moon bright bulging, proctoptosis.
2, the acquisition of RNA is organized
Use Trizol one-step method to extract total tissue RNA, read 260nm by Nanodrop ND-1000
With the purity that the absorbance (A) at 280nm measures RNA solution.Through 1% denaturing formaldehyde agarose gel electricity
Swimming, observes under ultraviolet transmission light, the integrity of detection RNA.
3, gene chip hybridization and scanning
After the linearized amplification of total serum IgE, cy3-UTP labelling, the cRNAs after fluorescent labeling uses RNEASY
Mini Kit purification, enters the cRNAs that labelling is good with the RNA Fragmentation Reagents of Amhion
Row fragmentation processes.Use people's full genome chip of expression spectrum (4x 44K gene) of Agilent company of the U.S.,
In chip hybridization stove, 65 DEG C of hybridization 17h, then eluting, dyeing, finally uses Agilent DNA
MicroarrayScanner scanner scanning.
4, chip data processes and analyzes
Chip after hybridization, after chip scanner reads data point, imports data to analyze software, for two groups
The natural logrithm absolute value of the ratio gene more than 2.0 or less than 0.5 is as difference expression gene.
5, statistical procedures
Using SPSS 13.0 statistical software to carry out data analysis, group difference compares employing one factor analysis of variance
Method, P < 0.05 difference has significant.
6, result
Chip results shows, filters out 654 differences between endometrial and normal endometrial tissue altogether
Different expressing gene, the gene 421 that wherein expression raises, the gene 233 that expression is lowered.
The difference expression gene filtered out verified by embodiment 2 large sample
Consider that yet there are no the gene carrying out studying about this gene and carcinoma of endometrium dependency in prior art makees
For candidate gene, considering the result of gene sequencing, (it expresses film in uterus to select UHRF1 gene simultaneously
Cancerous tissue raises) verify.
1, sample collection
Endometrial 50 example, normal endometrial tissue 60 example is collected according to the method for embodiment 1.
2, verify in mRNA level in-site
2.1 extract tissue RNA
Step is with embodiment 1.
2.2 reverse transcription
Use Reverse Transcriptase kit, with RT Buffer, l μ g total serum IgE is carried out converse record and synthesize cDNA.
Using 25 μ l reaction systems, each sample takes 1 μ g total serum IgE as template ribonucleic acid, in PCR pipe point
Do not add following components: DEPC water, 5 × RT Buffer, 10mmol/l dNTP, 0.1mmol/l DTT,
30 μm mol/l Oligo dT, 200U/ μ l MMLVRT, template ribonucleic acid.Hatch 1 hour for 42 DEG C, 72 DEG C 10
Minute, of short duration centrifugal.
2.3 PCR
Primer-design software Primer Premier 5.0 is used to design mRNA fluorescent quantitation upstream and downstream PCR primer,
Synthetic primer sequence, uses the operation of SYBR Green PCR Master Mix test kit, and concrete steps are normally
Bright book operates, and uses 25 μ l reaction systems, and each sample arranges 3 parallel pipes, all amplified reactions
Above to ensure the reliability of result.Prepare following reaction system (as shown in table 1), every
Operation is all carried out on ice:
The each component of table 1 quantitative fluorescent PCR and respective volume
With GAPDH as internal reference, using SYBR Green I as fluorescent marker, at Light Cycler fluorescence
The enterprising performing PCR of quantitative PCR apparatus reacts, and determines purpose band, Δ Δ CT by melt curve analysis analysis and electrophoresis
Method carries out relative quantification.
UHRF1 gene primer sequence is as follows:
Forward primer: 5 '-TATGAGGATGATGTGGAC-3 ' (SEQ ID NO.1)；
Downstream primer: 5 '-TGTTGGTGAGTTTCTGAT-3 ' (SEQ ID NO.2).
GAPDH gene primer sequence is as follows:
Forward primer: 5 '-TTTAACTCTGGTAAAGTGGATAT-3 ' (SEQ ID NO.3)；
Downstream primer: 5 '-GGTGGAATCATATTGGAACA-3 ' (SEQ ID NO.4)
2.4 result
Result as it is shown in figure 1, compared with normal endometrial tissue, UHRF1 base in endometrial
The mRNA level in-site of cause substantially raises, and difference has statistical significance (P < 0.05)
3, verify on protein level
3.1 extract tissue total protein
The operation of protein extraction is carried out according to the description of EpiQuik tissue/cell total protein extraction test kit.
3.2 Western blot detections
The protein quantification of extraction is carried out SDS-PAGE electrophoresis, carry out afterwards transferring film, closing, one anti-hatch,
Two anti-hatch, develop the color.
3.3 statistical procedures
Image J software is used to be analyzed the gray value of protein band, with β-actin as internal reference, will
The gray value of UHRF1 protein band is normalized.Result data is all with mean+SD
Mode represents, uses SPSS13.0 statistical software to carry out statistical analysis, and difference between the two uses
T checks, it is believed that when P < has statistical significance when 0.05.
3.4 result
Result as in figure 2 it is shown, compared with normal endometrial tissue, UHRF1 egg in endometrial
White level dramatically increases, and difference has statistical significance (P < 0.05).
Embodiment 3 suppresses UHRF1 gene expression
1, siRNA design synthesis
SiRNA sequence for UHRF1:
Positive-sense strand is 5 '-ACUCAUUGACCUUGUACAGCC-3 ' (SEQ ID NO.5)
Antisense strand is 5 '-CUGUACAAGGUCAAUGAGUAC-3 ' (SEQ ID NO.6)；
Above siRNA sequence and negative control siRNA sequence (siRNA-NC) are by Shanghai Ji agate pharmacy skill
Art company limited provides.
2, the cultivation of endometrial carcinoma cell and transfection
2.1 cells are cultivated
Ishikawa3-H-12 cell line all (contains in RPMI 1640 culture medium by the culture bottle adhere-wall culture of 50ml
10% hyclone, 100U/ml penicillin, 100g/ml streptomycin) in, at 37 DEG C, 5%CO2Saturated
The incubator of humidity is cultivated continuously.
2.2 cell transfecting
(1) first 24 hours are transfected, at 500 μ l nonreactive inoculation of medium 0.5-2*105Individual cell, during transfection
Cell degrees of fusion is 30-50%.Cell dissociation is mixed completely during bed board, it is to avoid cell accumulation grows.
(2) with 50 μ l serum-free medium dilution siRNA, (the final concentration of 33nM of transfectional cell, blows gently
Inhale 3-5 mixing.
(3) overturn mixing transfection reagent gently, dilute 1 μ l with 50 μ l serum-free mediums
Lipofectamine2000 3-5 mixing of pressure-vaccum, left at room temperature 5min gently.
(4) mixing transfection reagent and siRNA diluent, gently 3-5 mixing of pressure-vaccum, left at room temperature 20min.
(5), during transfection composite joins 24 porocyte plates, 100 μ l/ holes, jog cell plates mixing front and back is all
Even.
(6) cell plates are placed in 37 DEG C, 5%CO2Incubator is cultivated 18-48 hour.After transfecting 4-6 hour
Interchangeable fresh culture medium.
3, the jamming effectiveness of QPCR experiment detection siRNA is utilized
3.1 extract cell total rna utilizes conventional method to operate.
3.2 reverse transcription
Step is with embodiment 2.
3.3 QPCR
Step is with embodiment 2.
3.4 result
Result is as it is shown on figure 3, siRNA-UHRF1 can effectively suppress the expression of UHRF1 gene, poor
Different have statistical significance (P < 0.05).
The expression of the embodiment 4 UHRF1 gene mensuration to endometrial carcinoma cell multiplication capacity
1, step:
Application 5-bromo-2 ' Brdurds (Brd U) labelling and detection kit.Reference reagent box operation instruction,
Cell transfecting (transfection procedure is with embodiment 3), after 48 hours, is inhaled and is abandoned culture medium, adds the training of Brd U labelling
Foster base, 37 DEG C, 5%CO2Incubator is cultivated 60min.Discard culture medium, after PBS rinses, with 70%
Ethanol is fixed, overnight.Resisting the anti-Brd U antibody for mice with one, two resist the anti-mouse for band FITC
Fluorescent antibody, carries out immunoreation.Then flow cyctometry detection is carried out.
2, Brd U incorporation result:
Result shows: transfection siRNA-NC groups of cells incorporation efficiency average out to: (26.86 ± 1.43) %；
SiRNA-UHRF1 groups of cells incorporation efficiency average out to (10.12 ± 0.76) %, difference has statistical significance
(P<0.05).Above-mentioned test result indicate that, UHRF1 gene expression promotes the increasing of endometrial carcinoma cell
Grow.
In embodiment 5 endometrial carcinoma cell antibody and experiment
1, step:
Endometrial carcinoma cell Ishikawa3-H-12 is inoculated in 96 porocyte culture plates, every hole 2*103
Individual cells/well/200 μ l, is handled as follows after cell attachment:
Experimental group 1 (matched group): add unrelated monoclonal antibody (1:50) in endometrial carcinoma cell；
Experimental group 2: add anti-human UHRF1 monoclonal antibody (1:50) in endometrial carcinoma cell.
By cell at 37 DEG C, 5%CO2After incubator hatches 24 hours, add3H-TdR (1 μ Ci/ hole), then
Cultivate 24 hours, collect cell, add liquid scintillation solution, β calculating instrument detection cpm value.
2, statistical method
Experiment is all according to being repeated 3 times, and result data is all to come in the way of mean+SD
Representing, using SPSS13.0 statistical software to carry out statistical analysis, difference between the two uses t inspection,
Think when P < has statistical significance when 0.05.
3, result
Result as shown in Figure 4, compared to matched group, adds the groups of cells cell proliferation of anti-human UHRF1 monoclonal antibody
Slow down.Above-mentioned test result indicate that, the function of suppression UHRF1 albumen can suppress endometrial carcinoma cell to increase
Grow.
Embodiment 6 flow cytometer measures UHRF1 gene expression to apoptotic impact
The AnnexinV-FITC cell apoptosis detection kit buying BD company carries out apoptotic detection.
1, cell is cultivated and transfection
Step is with embodiment 3.
2, apoptosis detection
(1) the cell collected by trypsinisation not contained；
(2) with washed cell secondary (centrifugal collecting cell 5*105)；
(3) 500 μ l binding buffer suspension cells are added；
(4), after adding 5 μ l AnnexinV-FITC mixings, 5 μ l Propidium Iodide mixings are added；
(5) room temperature, lucifuge, reaction 5-15min；
(6) in 1h, observation and the detection of flow cytometer is carried out；
(7) with flow cytomery, excitation wavelength Ex=488nm launches wavelength Em=530nm.
3, statistical method:
Experiment is all according to being repeated 3 times, and result data is all to come in the way of mean+SD
Representing, using SPSS13.0 statistical software to carry out statistical analysis, difference between the two uses t inspection,
Think when P < has statistical significance when 0.05.
4, result:
Experimental result shows, transfection siRNA-NC groups of cells cell mean apoptotic rate is 12%, transfection
SiRNA-UHRF1 group cell mean apoptotic rate is 38%, and difference has statistical significance (P < 0.05).
The impact of embodiment 7Transwell experiment detection UHRF1 gene expression cell migration
Step:
1, from the refrigerator of-80 DEG C, frozen Matrigel thing is taken out, then mistake under 4 DEG C of temperature conditionss
Night so that it is become liquid.
2, take out 200 μ l serum-free cell culture mediums, add the Matrigel reagent of 50 μ l, at cryogenic conditions,
Preferably operate on ice face and be mixed evenly, respectively add 100 μ l, be placed on 37 DEG C, carbon dioxide
Incubator is cultivated 5h, the most often observes liquid case, when liquid becomes slightly white, illustrate that it has turned into
Solidification state.
3, cell (according to the embodiment 3 step operation) trypsinization that will transfect, with without serum
Culture medium is cleaned 3 times, then counts, then is made into cell suspending liquid.
4, gel is washed 1 time gently, then by 2*10 by the culture medium of serum-free5Cell suspension is in 100 μ l
In RPMI 1640, it is inoculated in room on transwell.
5, lower room adds 600 μ l 10%FBS RPMI 1640.
6, in 37 DEG C of incubators, after cultivating 12h, take out transwell cell, often organize all 4 samples of repetition.
7, wherein 1 cell discards culture medium, washes 3 times with the PBS without calcium, and 4% poly first ferment fixes 10min,
The cell that upper strata does not migrates wiped by cotton swab, and PBS washes 3 times, and violet staining or Giemsa dyeing, micro-
Microscopic observation.Remaining 3 cells to be digested with 0.25% membrane proteolytic enzyme by its lower floor's migrating cell, computation migration is thin
Born of the same parents' number.
Result:
Migration experimental result shows, transfects siRNA-NC groups of cells cell migration number average out to 228, turns
Dye siRNA-UHRF1 groups of cells cell migration number average out to 102, difference has statistical significance
(P<0.05)。
The explanation of above-described embodiment is only intended to understand the method for the present invention and core concept thereof.It is right to it should be pointed out that,
For those of ordinary skill in the art, under the premise without departing from the principles of the invention, it is also possible to the present invention
Carrying out some improvement and modification, these improve and modify also by the protection domain falling into the claims in the present invention.
Claims (10)
1. the product of detection UHRF1 gene or UHRF1 albumen is preparing diagnosis of endometrial carcinoma or prediction
Application in the instrument of endometrial carcinoma prognosis.
Application the most according to claim 1, it is characterised in that described detection UHRF1 gene or UHRF1
The product of albumen includes the product detecting the expression of UHRF1 gene or UHRF1 albumen.
Application the most according to claim 1 and 2, it is characterised in that described product includes can be in conjunction with
The nucleic acid of UHRF1 gene or can be in conjunction with the material of UHRF1 albumen；Described nucleic acid can detect UHRF1
The expression of gene；Described material can detect the expression of UHRF1 albumen.
Application the most according to claim 3, it is characterised in that described nucleic acid is to make in real-time quantitative PCR
The primer of specific amplified UHRF1 gene as shown in SEQ ID NO.1 and SEQ ID NO.2.
5. a diagnosis of endometrial carcinoma or the instrument of prediction carcinoma of endometrium prognosis, it is characterised in that described
Instrument includes the instrument that can detect the expression of UHRF1 gene or UHRF1 albumen.
Instrument the most according to claim 5, it is characterised in that described instrument includes can be in conjunction with UHRF1
The nucleic acid of gene or can be in conjunction with the material of UHRF1 albumen；Described nucleic acid can detect UHRF1 gene
Expression；Described material can detect the expression of UHRF1 albumen.
Instrument the most according to claim 6, it is characterised in that described nucleic acid is to make in real-time quantitative PCR
The primer of specific amplified UHRF1 gene as shown in SEQ ID NO.1 and SEQ ID NO.2.
8. the medicine treating carcinoma of endometrium, it is characterised in that described pharmaceutical pack containing UHRF1 gene or
The inhibitor of UHRF1 albumen.
Medicine the most according to claim 8, it is characterised in that described inhibitor can suppress UHRF1
Or relate to expression or the activity of the material of UHRF1 upstream or downstream pathway.
10. the application in the medicine of preparation treatment carcinoma of endometrium of the inhibitor described in claim 8 or 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610494676.XA CN105886660A (en) | 2016-06-29 | 2016-06-29 | Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610494676.XA CN105886660A (en) | 2016-06-29 | 2016-06-29 | Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105886660A true CN105886660A (en) | 2016-08-24 |
Family
ID=56719355
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610494676.XA Pending CN105886660A (en) | 2016-06-29 | 2016-06-29 | Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105886660A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107779509A (en) * | 2017-12-05 | 2018-03-09 | 天津脉络医学检验有限公司 | A kind of mark of pulmonary cancer diagnosis and its application |
| CN111363744A (en) * | 2020-02-26 | 2020-07-03 | 暨南大学 | Berberine-mediated UHRF1 gene inhibition and application thereof in preparation of medicines for treating multiple myeloma |
-
2016
- 2016-06-29 CN CN201610494676.XA patent/CN105886660A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 田侠等: "分子标志物在子宫内膜癌中的表达分析", 《现代生物医学进展》 * |
| 盛燕: "子宫内膜癌抑癌基因甲基化及UHRF1复合体表观遗传调控的研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
| 陈海燕: "子宫内膜癌中抑癌基因甲基化及其表观遗传调控的研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107779509A (en) * | 2017-12-05 | 2018-03-09 | 天津脉络医学检验有限公司 | A kind of mark of pulmonary cancer diagnosis and its application |
| CN111363744A (en) * | 2020-02-26 | 2020-07-03 | 暨南大学 | Berberine-mediated UHRF1 gene inhibition and application thereof in preparation of medicines for treating multiple myeloma |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105969901B (en) | Purposes of the MS4A6A as Huppert's disease diagnosis and treatment marker | |
| CN105886659A (en) | DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer | |
| CN105907879B (en) | Carcinoma of endometrium biomarker | |
| CN106222259B (en) | A kind of molecular marker of diagnosis and treatment Huppert's disease | |
| CN105886660A (en) | Application of UHRF1 (ubiquitin-like, containing PHD and RING finger domains, 1) as endometrial cancer diagnosis and treatment marker | |
| CN106011260B (en) | A kind of molecular marker of diagnosis and treatment carcinoma of endometrium | |
| CN106191283B (en) | The diagnosis and treatment product of Huppert's disease biomarker | |
| CN106011288B (en) | Huppert's disease diagnosis and treatment marker and its application | |
| CN107012256A (en) | Abdominal aneurvsm diagnosis and treatment mark | |
| CN105969904B (en) | Huppert's disease biomarker | |
| CN106011289A (en) | Application of LILRA2 gene and expression product thereof as diagnosis and treatment target of multiple myeloma | |
| CN106244705B (en) | Application of the ERC1 in preparation diagnosis or treatment hypopharyngeal cancer tool | |
| CN106119358B (en) | The diagnosis and treatment product of carcinoma of endometrium biomarker | |
| CN106947820A (en) | Purposes of the VCAN in adenocarcinoma of colon diagnosis and treatment | |
| CN106119357B (en) | Application of the TNS11 in preparation diagnosis and treatment carcinoma of endometrium product | |
| CN106011291B (en) | A kind of molecular marker of diagnosis and treatment hypopharyngeal cancer | |
| CN106011262A (en) | Endometrial cancer diagnosis and treatment marker and application thereof | |
| CN106381329B (en) | The diagnosis and treatment target of C18orf8 gene and its expression product as hypopharyngeal cancer | |
| CN106048031B (en) | Application of the SYNM in preparation diagnosis or treatment carcinoma of endometrium tool | |
| CN106148531A (en) | Hypopharyngeal cancer biomarker | |
| CN106011290A (en) | Application of RPL8 to preparation of tool for diagnosing or treating multiple myeloma | |
| CN106222257B (en) | Application of the IGSF6 in preparation diagnosis and treatment Huppert's disease product | |
| CN106947819A (en) | Adenocarcinoma of colon diagnosis and treatment mark | |
| CN106119399B (en) | Purposes of the ZNF385C as hypopharyngeal cancer diagnosis and treatment marker | |
| CN106011286B (en) | Hypopharyngeal cancer diagnosis and treatment marker and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: Room 1210, Building 3, Ronghua Xintai Building, 10 Ronghua South Road, Yizhuang Economic and Technological Development Zone, Daxing District, Beijing Applicant after: BEIJING MEDINTELL BIOMED Co.,Ltd. Address before: 100080 Beijing city Haidian District Shanyuan Street No. 1 cubic court building room 3103 Applicant before: BEIJING MEDINTELL BIOMED Co.,Ltd. |
|
| CB02 | Change of applicant information | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20200401 Address after: Room 2503, Qianshan building, building D2, Qingdao International Innovation Park Phase II, No. 1, Keyuan Weiyi Road, Laoshan District, Qingdao, Shandong Province Applicant after: Qingdao Yangshen biomedical Co.,Ltd. Address before: Room 1210, Building 3, Ronghua Xintai Building, 10 Ronghua South Road, Yizhuang Economic and Technological Development Zone, Daxing District, Beijing Applicant before: BEIJING MEDINTELL BIOMED Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160824 |
|
| RJ01 | Rejection of invention patent application after publication |