CN103954770A - Application of EVs-TRPC5 in test of breast cancer drug resistance degree - Google Patents
Application of EVs-TRPC5 in test of breast cancer drug resistance degree Download PDFInfo
- Publication number
- CN103954770A CN103954770A CN201410170932.0A CN201410170932A CN103954770A CN 103954770 A CN103954770 A CN 103954770A CN 201410170932 A CN201410170932 A CN 201410170932A CN 103954770 A CN103954770 A CN 103954770A
- Authority
- CN
- China
- Prior art keywords
- trpc5
- evs
- breast cancer
- cell
- drug resistance
- 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.)
- Granted
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
-
- 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
- G01N33/57415—Specifically defined cancers of breast
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/02—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by transmitting the radiation through the material
- G01N23/04—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by transmitting the radiation through the material and forming images of the material
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses application of EVs-TRPC5 in test of breast cancer drug resistance degree. A testing method comprises the following steps: (1) collecting blood samples of patients or animals who suffer from breast cancer, respectively removing whole cells, blood platelets and cell debris in the samples by means of centrifugation, and finally, centrifugally collecting EVs in the samples; and (2) testing an expression condition of TRPC5 in the collected EVs by means of RT-PCR or Elisa, wherein if the expression of the TRPC5 is positive, suspicious patients or animals with drug resistance are found out. According to the expression condition of TRPC5 in the blood EVs, the condition of the change of drug resistance of breast cancer tumor cells can be shown; therefore, the application of the EVs-TRPC5 in test of breast cancer drug resistance degree has the advantages of simple test method, high speed, high sensitivity and low cost.
Description
Technical field
The present invention relates to a kind of new purposes of TRPC5, the application of the transient receptor potential channel (TRPC5) that especially relates to extracellular vesica (EVs) in detecting breast cancer drug-resistant intensity.
Background technology
Breast cancer is the common cancer of women, and serious threat women is physically and mentally healthy.Chinese Anti-Cancer Association announces
New statistical figure show, since the nineties in 20th century, breast cancer incidence and mortality ratio all rise continually and steadily, and the incidence of disease is with annual 4.476% speed increment, and mortality ratio is with annual 3% speed increment.Chemotherapy is one of primary treatment means of breast cancer, and at present clinical commonly used drug has: (i) anthracycline, Japanese yew class, anti-metabolism, anti-microtubule class, alkylating agent, miscellany etc.; (ii) antiestrogenic, arimedex etc.; (iii) Herceptin (anti-Her-2), bevacizumab (anti-VEGF).
But according to American Cancer Society, 90% above tumor patient is died from chemotherapy resistance in various degree.Chemotherapeutics is produced to a great problem that resistance has become oncotherapy.Research through nearly decades, the resistance mechanism of finding at present has (1) cell membrane abc drug transport protein to express enhancing, reduce medicine absorption and increase medicine and discharge, P-glycoprotein (P-glycoprotein, P-gp) be that current most study is also of paramount importance abc drug transport protein, by MDRG, mdr1 encodes; (2) by making medicine generation alienation reduce pharmaceutically active, common enzyme has GST, CYP450 etc.; (3) drug target point mutation or expression variation etc.; (4) Tumor Heterogeneity; (5) tumor stem cell resistance etc.
Up-to-date, the effective method of early diagnosis tumour at present, is to find tumor marker by blood count.Tumor markers refers in the process of tumorigenesis, some intracellular matters of energy reacting cells canceration.By the detection to various tumor markerses, can do early diagnosis to corresponding tumour, detect curative effect and tumor recurrence situation clinically.Therefore, on the basis of further investigation resistance mechanism, exploitation breast cancer resistance molecular diagnosis index, method, realize reagent, reach that resistance is early stage, dynamic monitoring, most important for the diagnosis and treatment of breast cancer resistance in present stage, also for optimizing medicines structure design, the helpful reference of research and development drug resistance inversion drug provision, thereby there is great and real social value.
MUC1 is a kind of mucin, when normal cell is expressed, is a kind of transmembrane glycoprotein, under normal circumstances, on the epithelial cell top of mammary gland, intestines and stomach and urogenital tract, expresses, and glycosylation is complete.MUC1 degree normal epithelial plays lubricated and protective effect, mediation signal transduction and cell adhesion.
In breast cancer cell line mcf-7, by phosphorylation, MUC1 can participate in the signal transduction of receptor tyrosine kinase mediation in conjunction with Rrb/SOS, and tyrosine phosphorylation is the committed step that membrane receptor participates in signal transduction.The expression characteristic of breast cancer MUC1 comprises: high expressed, unconventionality expression are low; Glycosylation, high sialic acid; Top is decided to be unclear, and polarity is chaotic; Cytoplasm and cell surface have overexpression, and these molecules can enter serum from breast cancer cell.Have researcher to detect substrate sample patient with breast cancer, find high expressed MUC1, nearly 92% patient can detect the MUC1 expressing.
TRPC5 is transient receptor potential channel (transient receptor potential channels, TRP passage) family hypotype, it is the non-selective passage of the penetrating calcium ion of energy on cell membrane, mainly be distributed in brain, lung, testis and placenta, and mainly participate in the formation of growth cone and the growth of brain.This research department's research is found, transient receptor potential channel TRPC5 and tumor multi-medicine drug-resistant are closely related, TRPC5 occurs obviously to raise in the breast cancer cell (MCF-7/ADM) of adriamycin-resistant, and can indirectly mediate P-gp albumen that tumour the produces MDR expression in multidrug resistance tumor cells (as MCF-7/ADM).The applicant for this character application Patents, patent name be < < TRPC5 as drug target the application > > (patent No.: 201210318389.5) in reverse multiple drug resistance of tumor.
Extracellular vesica (extracellular vesicles, EVs) refers to the vesica that extracellular environment contains a large amount of movably cell membranes source.These EVs mainly comprise allochthon exosomes, microvesicle microvesicles and apoptotic body apoptotic bodies, and these dynamic EVs play vital effect in iuntercellular interchange and process of immune regulation.Lipid Rafts labelled protein flotillin-2 specifically expressing on EVs, can be used as Specific marker and is applied in research process.Tumour cell also produces EVs, and researcher finds to have a large amount of EVs to exist in the blood of glioblastoma, cancer of pancreas, cancer of the stomach and Patients with Acute Leukemia.In these EVs, include cell surface protein, RNA and DNA, by EVs, Vesicle-Containing is transported to recipient cell by donorcells, thereby exchange between mediated cell, and then cause the migration and invasion of tumour, yet the formation reason of these EVs structures and need further research with the correlativity of mammary cancer chemotherapy resistance.
Summary of the invention
The problems referred to above that exist for prior art, the applicant provides the application of a kind of EVs-TRPC5 in detecting breast cancer drug-resistant intensity.The present invention, according to the height of the expression of TRPC5 in blood EVs, can demonstrate the situation that breast cancer tumour cells resistance changes, and has advantages of that detection method is simple, speed is fast, highly sensitive, cost is low.
Technical scheme of the present invention is as follows:
The application of EVs-TRPC5 in detecting breast cancer drug-resistant intensity, concrete detection method is:
(1) collect the sick human or animal's who suffers from breast cancer blood sample, respectively by intact cell, blood platelet and cell fragment in centrifugal removal sample, the EVs in last centrifugal collection sample;
(2) method such as available RT-PCR, Elisa detects the expression of TRPC5 in the EVs collecting, positive if TRPC5 expresses, for there is the suspicious sick human or animal of drug resistance.
The technique effect that the present invention is useful is:
1, the present invention, by detecting the expression of TRPC5 in blood EVs, as an intermediate result, can tentatively obtain the resistance situation of tumor of breast.The expression of TRPC5 is higher, and the possibility that resistance occurs for the person that provides blood sample is also just higher.
2, the detection method that the present invention adopts is simple and quick, only needs to extract blood sample and just can detect, and has alleviated dramatically the misery of tumour patient;
3, by the present invention, can Preliminary detection go out patient in the resistance situation in this stage, combine other existing breast cancer resistance diagnostic techniques simultaneously, can be conducive to instruct doctor to propose therapeutic scheme more accurately, to obtain optimum curative effect.
Accompanying drawing explanation
Fig. 1 is that the TRPC5 of embodiment 1 expression on resistance breast cancer cell EVs detects schematic diagram;
Fig. 2 is that the TRPC5 of embodiment 2 detects schematic diagram at the xenograft tumor nude mice blood of resistance breast cancer and the expression in the tumor of breast patient's of chemotherapy blood EVs.
Embodiment
The material source or the preparation method that in embodiment, use are as follows:
1, cell
(1) wild type human breast cancer cell MCF-7/WT deposits storehouse (ATCC) purchased from US mode culture.
(2) multidrug resistance MCF-7/ADM is prepared and is preserved by Southern Yangtze University's drug design and molecular pharmacology research department, and its preparation method is as follows:
The wild type human breast cancer cell MCF-7/WT cell (purchased from ATCC) of newly recovery under cultivating normal condition, is cultivated in cell to 2 generation ~ 3 generations, make Growth of Cells stable, when cell converges, with pancreatin, digest and go down to posterity, second day upgrades nutrient culture media, 1/10 of the MCF-7/WT IC50 of simultaneously take adds adriamycin as initial concentration, after dosing second day, again change liquid, and the concentration that maintains adriamycin is carried out the routine cultivation of going down to posterity, after cytotostatic growth, improving drug concentration continues to cultivate, until cell can be stable growth in the nutrient culture media of 5 μ g/ml in doxorubicin concentration, obtain, whole preparation process is lasted 8 months.
2, antibody and siRNA
TRPC5 antibody (ab63151) is purchased from U.S. Abcam company; Goat polyclonal to rabbit IgG 15 nm Gold (ab27236) are purchased from U.S. Abcam company; TRPC5-siRNA is purchased from American I nvitrogen company.
3, patient
Blood sample picks up from 29 patient with breast cancers, and wherein 17 patients accepted anthracycline/taxone chemotherapy, and 12 patients did not accept any chemotherapeutics treatment.
4, experimental apparatus
Transmission electron microscope is purchased from Japanese Hitachi company; Laser confocal microscope is purchased from German Zeiss company; PCR instrument is purchased from U.S. Bio-Rad company; CO
2constant temperature cell culture incubator is purchased from U.S. Thermo company; Nucleic acid electrophoresis instrument system is purchased from U.S. Bio-Rad company.
Embodiment 1:TRPC5 expresses on the EVs of resistance breast cancer cell, and participates in the formation of EVs.
Method: collect MCF-7/ADM and the MCF-7/WT cell of in vitro culture, be prepared into transmission electron microscope sample, under transmission electron microscope, observe the textural difference of two kinds of cells, and Taking Pictures recording.Result as shown in Figure 1A.
The transmission electron microscope sample of MCF-7/ADM cell through BSA sealing, TRPC5 primary antibodie, immuno-gold labeling two is anti-hatch after, the distribution situation of observing TRPC5 under transmission electron microscope, and Taking Pictures recording.Result as shown in Figure 1B.
MCF-7/ADM cell is inoculated in the burnt capsule of copolymerization with 3000/hole, after cell attachment, carry out immunostaining, sample through BSA sealing, TRPC5 primary antibodie, green fluorescence two is anti-hatch after, the Expression and distribution situation of observing TRPC5 under laser confocal microscope, and Taking Pictures recording.Result as shown in Figure 1B.
The MCF-7/ADM cell of in vitro culture is first processed through Scrambled siRNA and TrpC5 siRNA, then be prepared into equally transmission electron microscope sample, under transmission electron microscope, observe the textural difference of MCF-7/ADM cell before and after TrpC5 siRNA disturbs, and Taking Pictures recording.Result as shown in Figure 1 C.
Result:
As shown in Figure 1A, transmission electron microscope results demonstration, compares with MCF-7/WT cell, and MCF-7/ADM surface of cell membrane has produced a large amount of EV structures.As shown in Figure 1B, immunofluorescence dyeing and immune transmission electron microscope results all show, TrpC5 a large amount on EVs is expressed.As shown in Figure 1 C, when MCF-7/ADM cell is respectively after Scrambled siRNA and TRPC5 siRNA process, after transmission electron microscope results shows that TRPC5 is suppressed, the EVs of MCF-7/ADM surface of cell membrane obviously reduces.These results show, compare with wild type MCF-7 cell, and the MCF-7 cell surface of adriamycin-resistant produces a large amount of EVs structures, and TRPC5 albumen is great expression in this structure, and participates in the formation of EVs.Because just TRPC5 can be detected in the EVs of mdr cell, illustrate that TRPC5 and resistance are corresponding.
Embodiment 2:RT-PCR method detects the expression of TRPC5 in blood sample EVs.
Method: inject respectively MCF-7/WT and MCF-7/ADM cell (5 * 10 at the flank of female nude mice
6individual cell/nude mice), cultivate 4 ~ 8 weeks, when tumour is grown to about 200mm
3time, at an ADM(3mg/kg of resistance tumor-bearing mice tumor locus injection in every three days), simultaneously at all tumor-bearing mice tumor locus TRPC5-siRNA of injection in every three days or contrast siRNA (40pmol), after 30 days, by cardiac puncture technology, collect blood and be kept in heparin sodium aqua, and collect EVs.
Collect the EVs in blood sample: plasma sample is collected in the polypropylene centrifuge tube containing EDTA, first with 300 * g, within centrifugal 10 minutes, remove intact cell, with 2500 * g, within centrifugal 20 minutes, remove blood platelet and cell fragment, repetitive operation 2 times, then under 4 degree conditions with centrifugal 1 hour of 16000 * g, centrifugal 1 hour of 100000 * g, be precipitated as EVs, with after PBS washing, be resuspended in PBS solution standby.
RT-PCR: use reverse transcription kit Reverse Transcriptase M-MLV (RNase H-) Kit (TaKaRa) that the genome reverse transcription of EVs is become to cDNAs, then detect by conventional RT-PCR method.Primer for detection of nude mice blood sample comprises:
TrpC5:?forward,?5′-GACCTGATAA?CCACTGAGAACCTGCTGAGC-3′;
reverse,?5′-GACAACCTCT?TGCCAAATGGGTCTTGATGC-3′;
flotillin2:?forward,?5′-CCAGAGACACTGTCCTTCCC-3′;
reverse,?5′-CATTGGAGCAAGGAGACAGAG-3′;
MUC1:?forward,?5′-TTCTTCCTGCTGCTGCTCCTCAC-3′;
reverse,?5′-GCACATCACT?CACGCTGACGTCTG-3′;
mdr1:?forward,?5′-CTTTCGAACTGCAAATATGCCTCC-3′;
reverse,?5′-GAGTTAGGAATGTAGCCCAGG-3′;
GAPDH:?forward,?5′-CAACGTGTCAGTGGTGGACC-3′;
reverse,5′-AGCAGTGAGGGTCTCTCTCTTC-3′.
Primer for detection of patient blood sample comprises:
TrpC5:?forward,?5′-AGACTTGCCATGGGCCACCTCTCATCAGAACC-3′;
reverse,?5-GAGGCGAGTTGTAACTTGTTCTTCCTGTCCATC-3′;
flotillin2:?forward,?5′-AGATCCGGCAGGAAGAGATT-3′;
reverse,?5′-GCTTCTGCCTTGAGCTTCAT-3′;
MUC1:?forward,5′-CGACTACTACCA?AGAGCTGCAGAGAGACAT-3′;
reverse,?5′-TGTAAGAGAGGCTGCTGCCA?CCATTACCTG-3′;
mdr1:?forward,?5′-CTGTTTGACTGCAGCATTGCTGAG?AACAT-3′;
reverse,?5′-CTGGCGCTTTGTTCCAGCCTGGACACTGAC-3′;
GAPDH:?forward,?5′-GGACTCATGACCACAGTCCATGCCATCACT-3′;
reverse,?5′-CTTGGAGGCCATGTGGGCCATGAGGTCCAC-3′.
Result: RT-PCR detects the mRNA level of each albumen in nude mice blood sample EVs, in 7 resistance tumor-bearing mices, the mRNA level of TrpC5, flotillin2, mdr1 and MUC1 all shows positive expression, the expression that shows the secretion of resistance tumor-bearing mice tumour has the EVs of TrpC5 to be released in the peripheral blood of nude mice, as shown in Figure 2 A.When drug-resistant tumor is after TRPC5 – siRNA processes, the transcriptional level of comparing TRPC5, flotillin2 and mdr1 with control group obviously declines, and statistics is shown in Fig. 2 A, and this result is further pointed out the formation of TRPC5 and EVs and transported closely related.
RT-PCR detects the mRNA level of each albumen in patient blood sample EVs, 17 in the patient of chemotherapy, the mRNA level of TRPC5, flotillin2, mdr1 and MUC1 all shows high expressed, and TRPC5 is expressed as strong positive, and in 12 patients without chemotherapy, the mRNA level of these albumen is all lower, as shown in Figure 2 B.This result has further shown that the EVs and the tumor drug resistance that carry TRPC5 have close contacting, and in prompting breast cancer disease human blood EVs, the expression of TRPC5 can be used as and detects index and be applied in the diagnoses and treatment of resistance breast cancer.
Embodiment 3:Elisa method detects the expression of TRPC5 in blood sample EVs.
Adopt the content of TRPC5 in this EVs of double antibody sandwich method (Elisa method) quantitative measurement blood sample.Use the Elisa detection kit for detection of humanized TRPC5 albumen business-like or that be manually coated with, detect sample for using the cracked blood EVs sample of protein lysate, according to conventional Elisa detection method, measure the content of TRPC5 in sample, by corresponding detecting instrument detection signal strength.TRPC5 in sample is more, and relatively detected value is just higher, and TRPC5 is just expressed as the positive, and the possibility that resistance occurs for the person that provides blood sample is so also just higher.
Claims (1)
- The application of 1.EVs-TRPC5 in detecting breast cancer drug-resistant intensity, is characterized in that concrete detection method is:(1) collect the sick human or animal's who suffers from breast cancer blood sample, respectively by intact cell, blood platelet and cell fragment in centrifugal removal sample, the EVs in last centrifugal collection sample;(2) available RT-PCR method or Elisa method detect the expression of TRPC5 in the EVs collecting, positive if TRPC5 expresses, for there is the suspicious sick human or animal of drug resistance.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410170932.0A CN103954770B (en) | 2014-04-25 | 2014-04-25 | EVs-TRPC5 is detecting the application in breast carcinoma resistance degree |
| PCT/CN2015/071054 WO2015165290A1 (en) | 2014-04-25 | 2015-01-19 | Uses of blood evs and associated molecules in tumor drug resistance detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410170932.0A CN103954770B (en) | 2014-04-25 | 2014-04-25 | EVs-TRPC5 is detecting the application in breast carcinoma resistance degree |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103954770A true CN103954770A (en) | 2014-07-30 |
| CN103954770B CN103954770B (en) | 2015-09-30 |
Family
ID=51332068
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410170932.0A Active CN103954770B (en) | 2014-04-25 | 2014-04-25 | EVs-TRPC5 is detecting the application in breast carcinoma resistance degree |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN103954770B (en) |
| WO (1) | WO2015165290A1 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015165290A1 (en) * | 2014-04-25 | 2015-11-05 | 江南大学 | Uses of blood evs and associated molecules in tumor drug resistance detection |
| CN106996971A (en) * | 2017-04-28 | 2017-08-01 | 江南大学 | Detect the kit and method of human peripheral EVs TRPC5 contents |
| CN107635571A (en) * | 2015-02-23 | 2018-01-26 | Md保健株式会社 | The composition of the treatment pregnancy related disorder of extracellular vesica containing the bacterium from bacillus |
| CN109153968A (en) * | 2016-04-12 | 2019-01-04 | 联合细胞Ev股份公司 | Extracellular vesica (EV) is separated from biologicfluid sample |
| CN115927202A (en) * | 2023-01-10 | 2023-04-07 | 北京爱思益普生物科技股份有限公司 | TRPC5 mutant cell strain and construction method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009052971A1 (en) * | 2007-10-26 | 2009-04-30 | Sanofi-Aventis | Complexes of trpc domains and sestd1 domains and methods and uses involving the same |
| WO2010056337A2 (en) * | 2008-11-12 | 2010-05-20 | Caris Mpi, Inc. | Methods and systems of using exosomes for determining phenotypes |
| CN101755208A (en) * | 2007-07-25 | 2010-06-23 | 路易斯维尔大学研究基金会公司 | Allochthon associated microRNA as diagnostic marker |
| CN102824637A (en) * | 2012-09-02 | 2012-12-19 | 江南大学 | Application of TRPC5 as drug target in reversing multi-drug resistance |
| CN102836430A (en) * | 2012-09-02 | 2012-12-26 | 江南大学 | Application of blocking antibody T5E3 to reversing of tumor multidrug resistance |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102121048A (en) * | 2010-01-11 | 2011-07-13 | 中山大学附属肿瘤医院 | POOL FQ-PCR (Fluorescence Quantitive Polymerase Chain Reaction) combined detection method of tumor drug resistance genes and kit thereof |
| CN103954770B (en) * | 2014-04-25 | 2015-09-30 | 江南大学 | EVs-TRPC5 is detecting the application in breast carcinoma resistance degree |
-
2014
- 2014-04-25 CN CN201410170932.0A patent/CN103954770B/en active Active
-
2015
- 2015-01-19 WO PCT/CN2015/071054 patent/WO2015165290A1/en active Application Filing
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101755208A (en) * | 2007-07-25 | 2010-06-23 | 路易斯维尔大学研究基金会公司 | Allochthon associated microRNA as diagnostic marker |
| WO2009052971A1 (en) * | 2007-10-26 | 2009-04-30 | Sanofi-Aventis | Complexes of trpc domains and sestd1 domains and methods and uses involving the same |
| WO2010056337A2 (en) * | 2008-11-12 | 2010-05-20 | Caris Mpi, Inc. | Methods and systems of using exosomes for determining phenotypes |
| CN102824637A (en) * | 2012-09-02 | 2012-12-19 | 江南大学 | Application of TRPC5 as drug target in reversing multi-drug resistance |
| CN102836430A (en) * | 2012-09-02 | 2012-12-26 | 江南大学 | Application of blocking antibody T5E3 to reversing of tumor multidrug resistance |
Non-Patent Citations (3)
| Title |
|---|
| XU SZ. ET AL.: "Generation of functional ion-channel tools by E3 targeting", 《NATURE BIOTECHNOLOGY》, vol. 23, no. 10, 18 September 2005 (2005-09-18) * |
| 蔡燕飞等: "耐阿霉素人乳腺癌细胞MCF-7/ADM钙稳态失调及其机制研究", 《中国药理学通报》, vol. 29, no. 6, 30 June 2013 (2013-06-30) * |
| 韩重阳等: "瞬时受体电位通道研究进展", 《生理科学进展》, vol. 39, no. 1, 31 December 2008 (2008-12-31) * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015165290A1 (en) * | 2014-04-25 | 2015-11-05 | 江南大学 | Uses of blood evs and associated molecules in tumor drug resistance detection |
| CN107635571A (en) * | 2015-02-23 | 2018-01-26 | Md保健株式会社 | The composition of the treatment pregnancy related disorder of extracellular vesica containing the bacterium from bacillus |
| CN107635571B (en) * | 2015-02-23 | 2021-09-28 | Md保健株式会社 | Composition for treating pregnancy-related diseases containing extracellular vesicles derived from bacteria belonging to the genus Bacillus |
| CN109153968A (en) * | 2016-04-12 | 2019-01-04 | 联合细胞Ev股份公司 | Extracellular vesica (EV) is separated from biologicfluid sample |
| CN106996971A (en) * | 2017-04-28 | 2017-08-01 | 江南大学 | Detect the kit and method of human peripheral EVs TRPC5 contents |
| CN115927202A (en) * | 2023-01-10 | 2023-04-07 | 北京爱思益普生物科技股份有限公司 | TRPC5 mutant cell strain and construction method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103954770B (en) | 2015-09-30 |
| WO2015165290A1 (en) | 2015-11-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2693938C2 (en) | Use of alphavirus in preparation of antitumor drugs | |
| CN103954770B (en) | EVs-TRPC5 is detecting the application in breast carcinoma resistance degree | |
| CN102770530A (en) | Catenae: serosal cancer stem cells | |
| CN106191067B (en) | Circular rna circ-NFATC3 and application thereof | |
| CN109762903A (en) | Application of the miR-1246 and/or TERF2IP in diagnosis and treatment glioma | |
| CN102510719A (en) | Treatment of brain metastases with inhibitors of endothelin receptors in combination with a cytotoxic chemotherapy agent | |
| CN107760784A (en) | Circular rna circ FOXP1 purposes | |
| CN107287174A (en) | Liver cancer marker OXCT1 and its application in diagnosing cancer of liver, treatment and prognosis | |
| CN104262459B (en) | Acute monocytic leukemia-associated antigen MLAA-34 epitope polypeptide, and vaccine and pharmaceutical application of acute monocytic leukemia-associated antigen MLAA-34 epitope polypeptide | |
| CN109453382A (en) | EphrinA1 albumen is preparing the application in the drug for inhibiting tumor cell invasion, transfer | |
| CN106701801B (en) | Detection marker and kit for B lymphoma and leukemia and application of detection marker and kit | |
| Li et al. | Elevated ITGA5 facilitates hyperactivated mTORC1-mediated progression of laryngeal squamous cell carcinoma via upregulation of EFNB2 | |
| CN103966334B (en) | The application of CSF2RB gene in prostate cancer with osseous metastasis | |
| CN109468380A (en) | IL1R2 is in Prognosis in Breast Cancer assessment and the application in targeted therapy | |
| CN111040032A (en) | Application of bidirectional regulator in preparation of preparation for diagnosing or regulating cell senescence and tumors | |
| CN109321656A (en) | Purposes of the protein D EPDC1 as the marker of the negative breast cancer of diagnosis three | |
| CN107868825A (en) | A kind of molecular marked compound of diagnosis and treatment adenocarcinoma of lung | |
| CN107326027A (en) | A kind of prna chimera body and application thereof | |
| Zhou et al. | Bone marrow derived mesenchymal stem cells involve in the lymphangiogenesis of lung cancer and Jinfukang inhibits the involvement in vivo | |
| CN104174012B (en) | The application of OTUD3 albumen in the product of preparation inhibition tumor growth | |
| CN110317878A (en) | A kind of long-chain non-coding RNA and its application for bladder cancer diagnosis and treatment monitoring | |
| CN104083368A (en) | Application of G-1 in preparation of G protein coupled receptor 30-based triple negative breast cancer targeting drugs | |
| CN103800919B (en) | TUFT1 application in preparing diagnosing cancer of liver and treatment preparation | |
| CN107144695B (en) | Application of the Arl13b albumen in cancer diagnosis | |
| CN104974988B (en) | Anti-pancreatic cancer monoclonal antibody and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |