WO2015165290A1 - Utilisation de vésicules extracellulaires (ve) du sang et de molécules associées en détection de pharmacorésistance d'une tumeur - Google Patents

Utilisation de vésicules extracellulaires (ve) du sang et de molécules associées en détection de pharmacorésistance d'une tumeur Download PDF

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WO2015165290A1
WO2015165290A1 PCT/CN2015/071054 CN2015071054W WO2015165290A1 WO 2015165290 A1 WO2015165290 A1 WO 2015165290A1 CN 2015071054 W CN2015071054 W CN 2015071054W WO 2015165290 A1 WO2015165290 A1 WO 2015165290A1
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evs
blood
drug resistance
expression
tumor
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Chinese (zh)
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马鑫
金坚
陈震
何冬旭
陈蕴
蔡燕飞
陆扬帆
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江南大学
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • 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

Definitions

  • the present invention relates to extracellular vesicles (EVs) and related molecules thereof, including cell membrane proteins, RNA and DNA, in particular, transient receptor potential channel (TRPC5), breast cancer resistance protein (BCRP), multidrug resistance
  • TRPC5 transient receptor potential channel
  • BCRP breast cancer resistance protein
  • MRP1 drug-related protein-1
  • Cancer also known as Malignant neoplasm, is a disease caused by a disorder that controls the growth and proliferation of cells. In addition to uncontrolled growth of cancer cells, cancer cells locally invade the surrounding normal tissues and even transfer to other parts of the body via the internal circulatory system or lymphatic system. Among various diseases, cancer is the "number one killer.” According to the World Health Organization report, there were approximately 12.7 million new cancer patients in the world in 2008, and 7.6 million died of cancer, especially in developing countries. The number of new cancer cases reached 56%. It is estimated that 2,020 years ago, the global annual 15 million cancer patients will be added.
  • drugs are: (i) anthracyclines, taxanes, antimetabolites, anti-microtubules, alkylating agents, miscellaneous, etc.; Antiestrogens, aromatase inhibitors, etc.; (iii) Trastuzumab (anti-Her- 2 ), bevacizumab (anti-VEGF).
  • P-glycoprotein is currently The most important and most important ABC drug transporter, encoded by the multidrug resistance gene mdrl; (2) reduce the drug activity by causing the drug to be catabolized, common enzymes include GST, CYP450, etc.; (3) drug target mutation or Changes in expression levels, etc.; (4) tumor heterogeneity; (5) cancer stem cell resistance.
  • Tumor markers refer to some intracellular substances that can respond to cancerous cells during tumorigenesis and development.
  • MUC1 is a mucin which is a transmembrane glycoprotein expressed in normal cells and is normally expressed at the apical end of epithelial cells of the breast, gastrointestinal tract and genitourinary tract, and glycosylation is complete. MUC1 normal epithelium provides lubrication and protection, mediates signal transduction and cell adhesion.
  • MUC1 By phosphorylation in breast cancer cell line MCF-7, MUC1 binds to Rrb/SOS and is involved in receptor tyrosine kinase-mediated signal transduction, whereas tyrosine phosphorylation is a membrane receptor involved in signal transduction A key step in the process.
  • the expression characteristics of breast cancer MUC1 include: high expression, low abnormal expression; glycosylation, high sialylation; apical unclear, polar confusion; overexpression of cytoplasm and cell surface, and these molecules will be from breast cancer The cells enter the serum.
  • TRPC5 is a subtype of the transient receptor potential channels (TRP channel) family, which is a non-selective channel on the cell membrane that is transparent to calcium ions, mainly distributed in the brain, lungs, testes and Placenta, and is mainly involved in the formation of growth cones and brain development.
  • TRP channel transient receptor potential channels
  • MCF-7/ADM adriamycin-resistant breast cancer cells
  • TRPC5 is up-regulated in adriamycin-resistant breast cancer cells (MCF-7/ADM) and can indirectly mediate tumors. Expression levels of multidrug-resistant P-gp proteins in multidrug resistant tumor cells (eg, MCF-7/ADM).
  • MCF-7/ADM multidrug-resistant P-gp proteins in multidrug resistant tumor cells
  • BCRP Breast cancer resistance protein
  • BCRP A transmembrane protein of 655 amino acid residues belonging to the ABC transporter superfamily. BCRP has only 6 transmembrane regions and 1 ATP binding site, so it is called incomplete transporter molecule. It is speculated that BCRP has the same composition. Dimers or heterodimers function as transmembrane channels. The expression of BCRP can be detected in placental syncytiotrophoblast cells, small intestine and colonic mucosal epithelial cells, bile duct membrane, mammary gland lobular and vascular endothelial cells and stem cells in human normal tissues. It is speculated that BCRP can inhibit certain exogenous absorption of digestive tract.
  • Substances include anticancer drugs and toxic substances participate in the formation of physiological functions such as the placental barrier.
  • BCRP is associated with clinical chemosensitivity in a variety of tumors including acute myeloid leukemia, non-small cell lung cancer, and breast cancer.
  • Multidrug resistance-associated protein-1 is mainly distributed in tissues such as lung, testis, and peripheral blood mononuclear cells, but can be detected in almost all tissues.
  • M RP1 has a molecular weight of 190 kD and consists of 1531 amino acid residues and is a glycosylated phosphoprotein integrated into the cell membrane.
  • the mRNA encoding MRP1 is approximately 6.5 kb, and a small portion of the sequence is homologous to P-glycoprotein mRNA, both of which are members of the superfamily of transporters of ATPase activity. Therefore, the tissue distribution and function of MRP is similar to that of P-glycoprotein. It is also used as a drug output pump.
  • ATP energy is used to remove drugs from cells.
  • Many natural drugs such as antibiotics, alkaloids and the like are substrates of MRP1, but MRP The effect does not appear to be inhibited by the MD R modulator.
  • Transfection of mrp gene into normal cells can confer resistance to many chemotherapeutic drugs, suggesting that overexpression of mrp gene can lead to MDR.
  • Extracellular vesicles refer to vesicles in which the extracellular environment contains a large number of mobile cell membrane sources. These EVs mainly include exosomes, microvesicles, and apoptotic bodies. These dynamic EVs play an important role in cell-to-cell communication and immune regulation.
  • the lipid raft marker protein flotillin-2 is specifically expressed on EVs and can be used as a specific marker in the research process. Tumor cells also produce EVs, and researchers have found large amounts of EVs in the blood of patients with malignant gliomas, pancreatic cancer, stomach cancer, and acute leukemia.
  • EVs contain cell membrane proteins, RNA and DNA, which transport vesicle contents from donor cells to recipient cells via EVs, thereby mediating cell-to-cell communication, which in turn leads to tumor migration and invasion.
  • RNA and DNA transport vesicle contents from donor cells to recipient cells via EVs, thereby mediating cell-to-cell communication, which in turn leads to tumor migration and invasion.
  • RNA and DNA transport vesicle contents from donor cells to recipient cells via EVs, thereby mediating cell-to-cell communication, which in turn leads to tumor migration and invasion.
  • RNA and DNA transport vesicle contents from donor cells to recipient cells via EVs, thereby mediating cell-to-cell communication, which in turn leads to tumor migration and invasion.
  • the formation of these EVs The reasons for its association with chemotherapy resistance in breast cancer remain to be further studied.
  • the Applicant has provided an application of EVS-TRPC5 in detecting the degree of tumor resistance.
  • the present invention can show the transformation of tumor cell resistance, and has the advantages of simple detection method, high speed, high sensitivity and low cost.
  • the tumor is a breast cancer or other type of tumor.
  • the related cell membrane protein refers to the transient receptor potential channel, TRPC5.
  • the related cell membrane protein refers to a breast cancer resistance protein, BCRP.
  • the related cell membrane protein refers to a multidrug resistance-associated protein-1, MRP1.
  • the present invention can initially obtain the drug resistance of breast tumors by detecting the expression level of related molecules in blood EVs as an intermediate result.
  • the detection method adopted by the invention is simple and rapid, and can be detected only by taking a blood sample, which greatly reduces the suffering of the tumor patient;
  • the patient's drug resistance at this stage can be initially detected, and the other existing tumor drug resistance diagnosis technology can help the doctor to propose a more accurate treatment plan to obtain the best. Efficacy.
  • FIG. 1 is a schematic diagram showing the detection of the expression of TRPC5 of Example 1 on drug-resistant breast cancer cells EVs;
  • FIG. 2 is a diagram showing the blood and the process of TRPC5 of Example 2 in xenograft nude mice with drug-resistant breast cancer. Chemotherapy of the breast Schematic diagram of the detection of expression in blood EVs of neoplastic patients;
  • FIG. 3 is a schematic diagram showing the detection of MRP1 expression in the drug-resistant breast cancer cell MVs of Example 4;
  • FIG. 4 is a schematic diagram showing the detection of the expression of BCRP in the drug-resistant breast cancer cell MVs of Example 4;
  • FIG. 5 is a schematic diagram showing the detection of MRP1 and Flotillin-2 in the drug-resistant breast cancer cell MVs of Example 4;
  • FIG. 6 is a diagram showing the detection of the expression of BCRP and Flotillin-2 in drug-resistant breast cancer cells MVs of Example 4;
  • FIG. 7 is a schematic diagram showing the expression of MRP1 of Example 5 in blood MVs of a chemotherapy-derived breast tumor patient;
  • Figure 8 shows the indication of the expression of BCRP in Example 5 in blood MVs of patients with chemotherapy-derived breast tumors.
  • Wild type human breast cancer cells MCF-7/WT was purchased from the American Type Culture Collection (ATCC).
  • Adriamycin-resistant human breast cancer cells MCF-7/ADM is prepared and preserved by the Department of Pharmaceutical Design and Molecular Pharmacology of Jiangnan University, and its preparation method is as follows:
  • the newly revived wild-type human breast cancer cell MCF-7/WT cells (purchased from ATCC) were cultured for 2 to 3 generations under normal cell culture conditions to stabilize cell growth, and the cells were confluent with trypsin. Digestion and passage, the medium was updated the next day, and doxorubicin was added at the same concentration as 1/10 of MCF-7/WT IC50. The drug was changed again after the second day of administration, and the concentration of doxorubicin was maintained. Conventional subculture, after the cells grow stably, increase the drug concentration and continue to culture until the cells can grow stably in the medium with the doxorubicin concentration of 5 g/ml. That is, the whole preparation process lasts for 8 months.
  • TRPC5 antibody (ab63151) was purchased from Abeam, USA; goat polyclonal to rabbit IgG 15 nm Gold (ab27236) was purchased from Abeam, USA; TRPC5-siRNA was purchased from Invitrogen, USA. [0040] 3, the patient
  • TEM Hitachi purchased from Japan; laser confocal microscope available from Zeiss, Germany; the PCR instrument was purchased from Bio-Rad Company; CO 2 cell incubator temperature purchased from Thermo Corporation; nucleic acid electrophoresis system available from U.S. Bio-Rad.
  • Example 1 TRPC5 is expressed on EVs of drug-resistant breast cancer cells and participates in the formation of EVs.
  • MCF-7/ADM cells were seeded in a confocal dish at 3000 cells/well, and immunostained after the cells were attached, and the samples were incubated with BSA, TRPC5-anti-green fluorescent secondary antibody, and The expression and distribution of TRPC5 were observed under a focused microscope and photographed. The result is shown in Fig. 1B.
  • MCF-7/ADM cells cultured in vitro were first treated with Scrambled siRNA and TrpC5 siRNA, and then prepared into transmission electron microscope samples. The structural differences of MCF-7/ADM cells before and after TrpC5 siRNA interference were observed under transmission electron microscope. Take a photo record. The result is shown in Figure 1C.
  • Example 2 RT-PCR was used to detect the expression of TRPC5 in blood samples.
  • MCF-7/WT and MCF-7/ADM cells (5 ⁇ 10 6 cells/nude mice) were injected into the flank of female nude mice, and cultured for 4-8 weeks.
  • ADM (3mg/kg) was injected every three days in the tumor site of drug-resistant tumor-bearing mice, and TRPC5-siRNA or control siRNA (40pmol) was injected every three days in the tumor site of all tumor-bearing mice, and passed after 30 days.
  • Cardiac puncture techniques collect blood stored in heparin sodium solution and collect EVs.
  • Plasma samples were collected in a polypropylene centrifuge tube containing EDTA, first centrifuged at 3000 xg for 10 minutes to remove intact cells, and centrifuged at 2500 xg for 20 minutes to remove platelets and cell debris, and repeated the operation twice. Then, it was centrifuged at 16000 x g for 1 hour at 16000 x g for 1 hour, and precipitated as EVs. After washing with PBS, it was resuspended in PBS solution for use.
  • RT-PCR Using Reverse Transcription Kit Reverse Transcriptase M-MLV (RNase H-) Kit
  • TrpC5 forward, 5'-GACCTGATAA CCACTGAGAACCTGCTGAGC-3';
  • flotillin2 forward, 5'-CCAGAGACACTGTCCTTCCC-3';
  • MUC1 forward, 5'-TTCTTCCTGCTGCTGCTCCTCAC-3';
  • mdrl forward, 5'-CTTTCGAACTGCAAATATGCCTCC-3';
  • GAPDH forward, 5'-CAACGTGTCAGTGGTGGACC-3';
  • Primers for detecting a blood sample of a patient include:
  • TrpC5 forward, 5'-AGACTTGCCATGGGCCACCTCTCATCAGAACC-3'; [0069] reverse, 5-GAGGCGAGTTGTAACTTGTTCTTCCTGTCCATC-3';
  • flotillin2 forward, 5'-AGATCCGGCAGGAAGAGATT-3';
  • MUC 1 forward, 5'-CGACTACTACCA AGAGCTGCAGAGACAT-3';
  • GAPDH forward, 5'-GGACTCATGACCACAGTCCATGCCATCACT-3';
  • RT-PCR was used to detect the mRNA level of each protein in EVs of nude mice.
  • the mRNA levels of TrpC5, flotillin2, mdrl and MUCl were positive, indicating drug resistance.
  • EVs expressing TrpC5 secreted by tumor-bearing mice were released into the peripheral blood of nude mice as shown in Fig. 2A.
  • the transcription levels of TRPC5, flotillin2 and mdrl were significantly decreased compared with the control group.
  • the statistical results are shown in Figure 2A. This result further suggests that TRPC5 is closely related to the formation and transport of EVs.
  • RT-PCR was used to detect the mRNA levels of various proteins in EVs of blood samples.
  • the mRNA levels of TRPC5, flotillin2, mdrl and MUCl were highly expressed, and TRPC5 expression was strongly positive, while 12 In patients without chemotherapy, the mRNA levels of these proteins are lower, as shown in Figure 2B.
  • the results further indicate that EVs carrying TRPC5 are closely related to tumor resistance, suggesting that the expression of TRPC5 in blood EVs of patients with breast cancer can be used as a detection index in the diagnosis and treatment of drug-resistant breast cancer.
  • Example 3 Elisa method was used to detect the expression of TRPC5 in EVs in blood samples.
  • the content of TRPC5 in blood sample EVs was quantitatively determined by the double antibody sandwich method (Elisa method).
  • the commercially available or manually coated Elisa assay kit for the detection of human TRPC5 protein is used to detect samples of blood EVs lysed with protein lysate, and the content of TRP C5 in the sample is determined according to the conventional Elisa assay. , use the corresponding detection instrument to detect the signal strength.
  • the more TRPC5 in the sample the higher the relative detection value, and the higher the TRPC5 expression, the more likely the person who provides the blood sample will become resistant. High.
  • MRP1 antibody (ab24102) and BCRP antibody (ab3380) were purchased from Abeam, USA; goat anti-MUCl monoclonal antibody (sc-6825) was purchased from Santa Cruz, USA; Alexa Fluor 488 labeled Donkey Anti-Mouse IgG (H+ L) Antibody and Alexa Fluor 546-labeled donkey anti-goat IgG (H+L) Antibody was purchased from Invitrogen, USA.
  • the tumor samples were selected before surgery for 2-4 chemotherapy courses or preoperative untreated tumors. Blood samples were taken from 55 breast cancer patients, including 34 patients receiving 3-6 cycles after surgery. Patients with taxoid-like adjuvant chemotherapy (extracted before the next cycle) and 21 patients who did not receive chemotherapy before surgery.
  • Laser confocal microscope was purchased from Zeiss, Germany; PCR instrument was purchased from Bio-Rad, USA; CO 2 thermostatic cell culture incubator was purchased from Thermo Corporation, USA; nucleic acid electrophoresis system was purchased from Bio-Rad, USA; flow cytometry The instrument was purchased from BD Corporation of the United States.
  • Example 4 FISH method was used to detect the expression of MRP1 and BCRP in MVs of blood samples.
  • Method The patient tumor cells were inoculated into a confocal dish at 3000 cells/well, and immunostained after the cells were attached, and the samples were incubated with BSA blocking, MRP1/BCRP-anti, red fluorescent secondary antibody, and the nuclei were incubated. After counterstaining with PADI, the expression and distribution of MRP1 and BCRP were observed under laser confocal microscopy and photographed. The results are shown in Figure 3 and Figure 4.
  • the patient tumor cells were seeded in a confocal dish at 3000 cells/well, and immunostained after the cells were attached, and the samples were incubated with BSA blocking, MRP1/BCRP and Flotillin-2 anti-green fluorescent secondary antibody, and After counterstaining the nucleus with PADI, the expression and distribution of MRP1, BCRP and Flotillin-2 were observed under laser confocal microscopy and photographed. The results are shown in Figure 5 and Figure 6.
  • Example 5 RT-PCR method, flow cytometry was used to detect the expression of MRP1 and BCRP in MVs of blood samples.
  • Plasma samples were collected in polypropylene tubes containing EDTA, first centrifuged at 300 ⁇ g for 10 minutes to remove intact cells, centrifuged at 2500 ⁇ g for 20 minutes to remove platelets and cell debris, and repeated operations twice. Then, after centrifugation at 16000 xg for 1 hour, centrifuge at 16000 xg for 1 hour, precipitate into MVs, wash with PBS, and resuspend in PBS solution for use.
  • RT-PCR Using Reverse Transcription Kit Reverse Transcriptase M-MLV (RNase H-) Kit
  • TaKaRa Reverse transcribes the genome of MVs into cDNAs, which are then detected by conventional RT-PCR methods.
  • Primers for detecting a blood sample of a patient MRP1 include:
  • GAPDH forward, 5'-CTCCTGCACCACCACCAACTGCTTAGC-3';
  • MRP1 forward, 5'-CTGCACGACCTCCGCTTCAAGAT-3';
  • Flotillin-2 forward, 5'-AGATCCGGCAGGAAGAGATT-3';
  • MUC1 forward, 5 ' -CGACTACTACCAAGAGCTGCAGAG AGACAT-3 ';
  • Primers for detecting a blood sample of a patient BCRP include:
  • BCRP forward, 5'-CAG CCG TGG AAC TCT TTG TGG TAG AGA AG-3';
  • Flotillin-2 forward, 5,-AGATCCGGCAGGAAGAGATT-3,; [0116] Reverse, 5 ' -GCTTCTGCCTTGAGCTTCAT-3 '
  • MUCl forward, 5 ' -CGACTACTACCAAGAGCTGCAGAG AGACAT-3 '; [0118] Reverse, 5 ' -TGTAAGAGAGGCTGCTGCCACCATTACCTG-3 ' .
  • GAPDH forward. 5,-CTC CTG CAC CAC CAA CTG CTT AGC-3';
  • MVs were incubated at 4 degrees and then incubated with BSA-blocked, MRP1/BCRP and M UC 1 primary antibodies, Alexa Fluor 488 and Alexa Fluor 546 labeled secondary antibodies, using flow cytometry Detection.
  • mRNA levels of MRP1, BCRP, Flotillin-2, MUCl and GAPDH showed high expression (A curve of Fig. 7 and Fig. 8), and MRP1 and BCRP expression were strongly positive (B curves of Fig. 7 and Fig. 8).
  • the mRNA levels of these proteins were lower in 21 patients without chemotherapy.
  • MRP1 or BCRP in blood MVs of breast cancer patients can be used as a detection index in the diagnosis and treatment of drug-resistant breast cancer.
  • breast cancer tumors were selected as samples, and cell membrane proteins, RNA and DNA (TRPC5, BCRP, MRP1, Flotillin-2) specifically expressed in EVs of breast cancer cells were selected, by RT-PCR, flow Cell technology, or Elisa method to detect their specific expression, according to the level of expression, indicating the possibility of multidrug resistance in breast cancer.
  • RNA and DNA the amount of expression of these substances was measured by the same method as described above, thereby demonstrating the possibility of multidrug resistance.
  • the expression molecules used in the examples are all molecules related to multidrug resistance of all tumors, therefore, The method provided by the application is universal.

Abstract

L'invention concerne des utilisations de vésicules extracellulaires (VE) du sang et de molécules associées en détection de pharmacorésistance d'une tumeur. L'invention concerne un procédé de détection particulier qui comporte : (1) le prélèvement d'échantillons de sang sur des patients ou des animaux souffrant de tumeurs, l'élimination, par centrifugation, des cellules complètes, des plaquettes et des débris cellulaires présents dans les échantillons et le recueil, par centrifugation, de VE présents dans les échantillons ; (2) la détection, à l'aide d'un procédé de chimiluminescence, d'un procédé de RT-PCR, de cytométrie en flux ou d'un procédé ELISA, d'une expression de protéine membranaire cellulaire, d'ARN ou d'ADN apparentés dans les VE recueillis. Si l'expression est positive, un patient ou animal supposé présenter une pharmacorésistance est trouvé. Les molécules associées aux VE comprennent TRPC5, BCRP ou MRP1.
PCT/CN2015/071054 2014-04-25 2015-01-19 Utilisation de vésicules extracellulaires (ve) du sang et de molécules associées en détection de pharmacorésistance d'une tumeur WO2015165290A1 (fr)

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