CN113862362A - Application of testis specific expression protein ARAP1 in diagnosis and treatment of breast cancer - Google Patents
Application of testis specific expression protein ARAP1 in diagnosis and treatment of breast cancer Download PDFInfo
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Abstract
The invention relates to the field of biomedicine. The invention belongs to the technical field of medical detection, and particularly relates to a marker for growth, invasion and transfer capacity of breast cancer cells and application thereof. ARAP1, only testis tissue expresses the protein in normal tissue, but the expression level of ARAP1 in breast cancer is obviously increased. The present invention discloses for the first time that ARAP1 can interact with the androgen receptor AR. Research on the action mechanism of the ARAP1 shows that the transcriptional activity of an androgen receptor AR can be remarkably reduced, and ARAP1 silencing can remarkably activate an anti-cancer gene downstream of the AR. The functional research of the ARAP1 provides a new idea and a new scheme for treating the breast cancer. Has important clinical value for the targeted drug development and targeted silencing of the ARAP 1.
Description
Technical Field
The invention belongs to the technical field of medical detection, and particularly relates to a marker for growth, invasion and transfer capacity of breast cancer cells and application thereof.
Background
Androgen Receptor (AR) is expressed in about 60% of cases in ER-positive breast cancer, and in recent years, more and more studies have shown that the regulation of AR downstream gene expression by AR transcriptional co-regulators is critical in suppressing breast cancer proliferation and endocrine resistance. Therefore, the identification and analysis of epigenetic mechanism of AR co-regulatory factor in AR mediated gene transcription are receiving more and more attention, and the method has important scientific significance for further research of potential drug target in tumor treatment.
Female breast cancer is a malignant tumor which occurs in female mammary epithelial cells, the incidence rate of the malignant tumor is counted to exceed that of lung cancer in the first place and the mortality rate of the malignant tumor is counted in the second place in 2020. Estrogen Receptor (ER) positive breast cancer accounts for 75% of the total breast cancer cases in clinical patients according to molecular typing. Because of the positive expression of ER, such patients often use neoadjuvant endocrine therapy targeting the ER-E2 signaling pathway to ameliorate the progression of cancer. Clinical studies have found that nearly one-third of patients present with intrinsic endocrine therapy resistance, and half of patients develop distant metastasis and malignant proliferation after undergoing chronic endocrine therapy. Irreversible endocrine drug resistance is generally present at the end of treatment for ER-positive breast cancer.
Because of the high heterogeneity of breast cancer, the treatment regimens vary greatly from patient to patient and from stage to stage within the same patient, and early detection and typing of breast cancer is important. However, few molecular markers related to early ER positive breast cancer diagnosis exist, so that the disease progression and prognosis of ER positive breast cancer are difficult to judge, and the condition delay is delayed.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a novel protein ARAP1 capable of interacting with androgen receptor AR, and researches show that the protein is only expressed in testis tissues in normal tissues. Research on the action mechanism of the ARAP can find that the ARAP1 can obviously down-regulate the transcriptional activity of AR, and ARAP1 silencing can obviously up-regulate the cancer suppressor gene downstream of AR.
In order to achieve the above object, the present invention provides the following technical solutions.
The invention provides an application of a reagent for detecting the expression level of an ARAP1 gene or an encoding protein thereof in preparing a kit for diagnosing and screening breast cancer.
Further, the ARAP1 gene or the protein encoded by the same exhibits high expression in breast cancer cells.
Further, the application detects the expression level of the ARAP1 gene or the coding protein thereof in the sample through RT-PCR, real-time quantitative PCR, in-situ hybridization, Northern blotting, a chip, a high-throughput sequencing platform, immunohistochemistry or enzyme-linked immunosorbent assay; the sample is tissue, serum or cells.
Further, the application contains a specific primer for amplifying the ARAP1 gene, a probe hybridized with the nucleotide sequence of the ARAP1 gene or an antibody specifically bound with the ARAP1 protein.
Further, the specific primer sequences for amplifying the ARAP1 gene are as follows:
ARAP1-F：5’-AGAGTCCCAGGCAGGACAA-3’；
ARAP1-R：5’-GCAGAAGGCTGAGGAACACT-3’；
the antibody is a monoclonal antibody or a polyclonal antibody.
The use of any one of the preceding claims, wherein the breast cancer is ER-positive breast cancer.
The invention also provides application of the expression inhibitor of the ARAP1 gene in preparing a medicament for treating breast cancer.
Further, the targeting sequence of the ARAP1 gene inhibitor is as follows: 5'-TGGATTTGTACCATTCTTCT-3' are provided.
Further, the inhibitor includes siRNA or shRNA containing the target sequence.
Further, the medicament comprises a pharmaceutically acceptable carrier and/or excipient.
Compared with the prior art, the invention has the beneficial effects.
Androgen Receptor (AR) is expressed in about 60% of cases in ER-positive breast cancer, and AR can change the location of ER on chromatin in ER-positive breast cancer so as to directly influence the transcriptional activity of ER downstream cancer-promoting genes and further play a role in inhibiting cancer. Androgens, known as ligands for AR, have also been used in clinical treatment of ER-positive breast cancer, suggesting our role of androgens and androgen receptors in cancer suppression in ER-positive breast cancer.
Compared with the prior art, the method disclosed by the invention applies modern molecular biology technologies such as network data, a protein immunoblotting experiment, a protein immunoco-precipitation experiment, a GST-Pulldown experiment, an immunofluorescence co-localization experiment, a luciferase double-gene reporting experiment, a real-time quantitative PCR experiment, a cell growth curve experiment, a cell transwell experiment and the like. The targeted ARAP1 gene interference siRNA is successfully constructed, and the influence on the growth and the invasion and transfer capacity of the breast cancer cells after the interference of the ARAP1 gene is detected at the cell level.
The invention discovers that the AR interaction protein ARAP1 is related to proliferation and invasion and metastasis of breast cancer cells for the first time, and the expression quantity of the AR interaction protein ARAP1 reflects the occurrence and development degree of breast cancer to a certain extent, thereby providing a new idea and a new scheme for treating the breast cancer.
Drawings
FIG. 1ARAP1 specific expression study.
Figure 2 ARAP1 interacts with the androgen receptor AR.
Figure 3ARAP1 down-regulates AR-mediated gene transcription.
Figure 4 silencing of ARAP1 can promote cancer cell proliferation.
Fig. 5 silencing of ARAP1 significantly promoted invasion and metastasis of breast cancer cells.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the equipment and reagents used in the examples and test examples are commercially available without specific reference. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.
The invention discloses a molecular marker for detecting the growth and invasive metastatic capacity of breast cancer cells and application thereof, which are shown in the following embodiments. The breast cancer cell lines involved in the present invention: MCF7, purchased from shanghai department of sciences cell bank. The ARAP1 antibody was purchased from Cellsignalttechnology, Inc. The real-time quantitative PCR method refers to TIANGEN instruction to extract total RNA of cells, after concentration measurement, reverse transcription is carried out according to a Prime script RT Master Mix kit of TaKaRa, and related gene expression is detected according to a SYBR Select Master Mix kit of appliedbiosystems (ACTB is an internal reference). Synthesis of siRNA and shRNA targeting ARAP1 was purchased from gecky corporation. The transfection method is carried out according to the specification of a LipofectamineRNAIMAMAX Reagent transfection Reagent of Invitrogen company, and expression and interference effects are detected by a q-PCR and Western blot method. Cell migration experiment Transwell (8mm pore size) chambers were purchased from Cosar and starved for culture. Cell scoring experiments 12-well plates were used and purchased from Cosar.
Example 1 cell recovery, culture, transfection and protein co-immunoprecipitation assay (co-IP) the interaction of the ARAP1 protein with the androgen receptor AR protein in the MCF7 cell line was examined.
Cell recovery: the MCF7 cells were removed from the liquid nitrogen and immediately placed in a 37 ℃ water bath for rapid thawing to minimize cell timeAnd 4, completely thawing the mixture. After the surfaces of the frozen tubes were sterilized by wiping with 75% alcohol, they were transferred to previously prepared 5ml LEP tubes (0.9 ml of DMEM + 10% FBS had been previously added) and centrifuged at 800rpm for 5 min. Discarding supernatant, resuspending with 1mL DMEM complete medium (DMEM + 10% FBS), mixing, adding 5mL DMEM complete medium into culture flask, transferring the resuspended cell suspension to culture flask, placing at 37 deg.C, and 5% CO2Culturing in an incubator, replacing the culture medium once the next day, continuing culturing, and beginning to passage when the cell density is about 80-90%.
Cell subculturing: discarding old culture solution in culture flask with cell density of about 80% -90%, washing with preheated PBS for 1-2 times, adding 1mL trypsin digestive juice (0.25% pancreatin + 0.02% EDTA) into the culture flask, digesting at 37 deg.C for 2-3min, observing cell state under microscope, adding 1mL LDMEM complete culture medium to stop digestion when cell volume is reduced and cell gap is enlarged, and blowing and beating the bottom of the flask gently and repeatedly with pipette. Collecting the blown cell suspension into a 5mL centrifugal tube, centrifuging at 900rpm for 5min, discarding supernatant, resuspending with 1mL MEM complete medium, mixing, adding 5mL MEM complete medium into a new culture flask, transferring appropriate amount of resuspended cell suspension into the culture flask, placing at 37 deg.C, and adding 5% CO2Culturing in an incubator. After the cell density reaches 80% -90%, F2 cells are inoculated to a 100mm culture dish for culture according to the passage method.
Cell transfection: when the cell density of the 12-well plate is about 30% -50%, taking ARAP1 plasmid with FLAG label as an experimental group, taking 3 EP tubes, marking A, B, C, adding 5 mu g of FLAG-ARAP1 into A, adding 5 mu g of FLAG-ARAP1+2 mu g of AR into B, C, gently mixing uniformly, and standing for 15 minutes at room temperature; adding the complex into the culture medium of 100mm culture dish inoculated with cultured cells in the above step, respectively, placing at 37 deg.C and 5% CO2And (3) continuously culturing in the incubator, changing the solution 4-6h after transfection, adding Dihydrotestosterone (DHT) with the final concentration of 10nM into A, C group, adding absolute ethyl alcohol with the same volume into B group, continuously culturing for 48h, and then carrying out other detection steps after transfection.
Protein co-immunoprecipitation experiments: the cells were washed 2 times with ice PBS, collected into a 1.5mL centrifuge tube with a cell scraper, centrifuged at 900rpm for 5min, and the supernatant was discarded. Cells were lysed on ice for 30min using 500 μ L of cell lysate (495 mL TNE +5 μ L100 x protease inhibitor), and then 3 times per tube for 5 seconds using ultrasound. After centrifugation at 12000rpm for 20min, the supernatant was transferred to a fresh siliconized centrifuge tube. 10% was split per tube as input, leaving 450 μ L of cell lysate pre-cleared: add 40. mu.L of protein beads and rotate at 4 ℃ for 1 hour. After centrifugation the beads were discarded and the supernatant was transferred to a fresh siliconized centrifuge tube. Add 80. mu. Lbeads to each tube, add 5. mu.L of FLAG antibody, and spin at 4 ℃ overnight. The following day at 1500rpm, centrifuge for 5min and discard the supernatant, wash beads3 with TNE for 10min each. Finally, 2 × LoadingBuffer 60 μ L of resuspension beads was used for future use.
Western Blot: (1) preparing polyacrylamide gel: 8 percent polyacrylamide gel is prepared according to the molecular weight of the protein. Cleaning a 1.5mm glass plate for preparing glue, airing, aligning, placing into a clamp, clamping, vertically clamping on a frame, preparing a glue preparation reagent according to a specification, adding separation glue firstly, quickly and slowly to avoid generating bubbles, adding absolute ethyl alcohol when adding to a position 1.5cm away from the upper end, sealing, standing for 20 minutes, removing the absolute ethyl alcohol, cleaning for 3 times by using ionized water, adding concentrated glue to the top end, inserting a comb, paying attention to the fact that no bubbles are generated, standing for about 10 minutes, and loading. (2) Loading and electrophoresis: and adding 20ul of prepared samples into each hole in a certain sequence, separating the periphery of each group of experimental samples by a pre-dyed protein Marker, wherein the initial voltage is 80V for about 40min, changing the voltage to 120V for about 1.5h after bromophenol blue enters separation gel, and stopping electrophoresis after the micromolecular protein is separated. (3) Film transfer: activating a PVDF membrane with a proper size in formaldehyde for about 1min, covering the gel after electrophoresis with the PVDF membrane, covering filter paper and sponge on non-contact surfaces of the PVDF membrane and the gel, and placing the gel in an electric rotating tank for 90min at a constant current of 300 mA. (4) And (3) sealing: after electrotransfer, the PVDF membrane was removed and placed in the freshly prepared blocking solution (1 XPBST containing 5% skimmed milk) on a shaker at room temperature for 2 h. (5) Sealing a first antibody: after the blocking is finished, 1 XPBST diluted primary antibody (anti-FLAG antibody is diluted 2000 times, anti-AR antibody is diluted 1000 times, each dilution is 2mL put into a centrifuge tube) containing 5% skim milk is prepared, the primary antibody is cut at the position of the corresponding protein, 2mL diluted corresponding primary antibody is respectively added into an antibody sealing box, and the mixture is kept on a shaker at 4 ℃ overnight or at room temperature for 2 h. (6) Washing the membrane, hatching the secondary antibody, washing the secondary antibody: the PVDF membrane is taken out and washed 3 times on a shaking bed of 1 XPBST, each time for 15min, the washed membrane is respectively placed in an antibody sealing box, 1 XPBST diluted secondary antibody (the secondary antibody is diluted by 5000 times and diluted by 2mL in a centrifuge tube) containing 5% of skimmed milk is prepared, 2mL of the diluted corresponding secondary antibody is respectively added in the antibody sealing box, the temperature on the shaking bed is 1h, and the membrane is washed 3 times on the shaking bed with 1 XPBST, each time for 10 min. (7) And (3) developing: mixing ECL luminous liquid A, B liquid in equal proportion according to the specification of a chemiluminescence kit (prepared before use), sucking the washed membrane to dry redundant 1 xPBST by using absorbent paper, placing the front side of the membrane on a luminescent plate upwards, uniformly dripping developing solution, and taking a picture by a gel imaging system and storing.
As a result, ARAP1 in MCF7 cells could interact with AR as shown in FIG. 2-A.
Example 2 real-time quantitative PCR detection of the mRNA interference effect of ARAP1 and the effect on target genes downstream of AR in cell lines.
Cell subculturing: discarding old culture solution in culture bottle with cell density of about 80% -90%, washing with preheated PBS for 1-2 times, adding 1mL trypsin digestive juice (0.25% pancreatin + 0.02% EDTA) into the culture bottle, digesting at 37 deg.C for 2-3min, observing cell state under microscope, adding 1mL DMEM complete culture medium to stop digestion when cell volume is reduced and cell circle becomes larger, and blowing the bottom of the bottle gently and repeatedly with pipette. Collecting the blown cell suspension into a 5mL centrifugal tube, centrifuging at 800rpm for 5min, discarding the supernatant, re-suspending with 1mL DMEM complete medium, mixing, adding 5mL DMEM complete medium into a new culture flask, transferring the re-suspended cell suspension to the culture flask, placing at 37 deg.C and 5% CO2Culturing in an incubator. After the cell density reaches 80% -90%, F2 cells are inoculated to a 6-well plate for culture according to the passage method.
Cell infection: when the cell density of the 6-well plate is about 30% -50%, the SHARAP1 virus is used as an experimental group, the shCtrl virus is used as a control group, and the cells are infected by standardized operation according to the Kjeldahl gene virus infection manual. After 24 hours, the medium was changed and the selection was carried out using puromycin at a final concentration of 50nM, and after the cells had grown to 80% -90% density, the F3 cells were plated onto 35mm dishes as described above. After the cells are full, the next operation can be carried out.
Extraction of total RNA: after cell collection, total cellular RNA was extracted according to TIANGEN instructions, during which rnase-free procedures were followed, wearing sterile gloves and masks. The method comprises the following specific steps: (1) the RL lysate (added with beta-mercaptoethanol) is used for lysing cells, 70% ethanol is prepared in advance, and the RL lysate and the 70% ethanol are marked separately to avoid confusion; (2) adding 350 μ L of prepared RL cell lysate into each well, slightly stroking the bottom of a gun head in clockwise and counterclockwise directions to fully lyse cells, slowly dripping the lysed solution onto a CS filter column along the side wall, clearly marking, rapidly centrifuging in a centrifuge, and collecting filtrate (2min, 4 ℃, 12000 rpm); (3) slowly adding 70% ethanol into the filtrate, slowly adding about 350 μ L of ethanol into each hole, gently mixing, transferring the mixed solution into a CR3 adsorption column, placing in a corresponding collection tube, centrifuging again (90s, 4 deg.C, 12000rpm) and discarding the filtrate, and placing the adsorption column into the collection tube again; (4) adding 1350 μ L deproteinized solution RW into each adsorption column along the side wall, adjusting the rotation speed of a centrifuge to 12000rpm, keeping the temperature at 4 ℃, centrifuging for 60s, removing filtrate, and placing the adsorption column into a collection tube; (5) suspending and dripping 80 μ L of DNase I working solution from the middle of each column, taking care not to drip to the side wall, and standing for 15min at room temperature; (6) slowly adding deproteinizing solution RW 1350 μ L along the side wall of each column, centrifuging at 4 deg.C in a high-speed centrifuge at 12000rpm for 60s, discarding the filtrate, and replacing the adsorption column with the collection tube; (7) adding 500 μ L RW rinse solution, standing for 2min, centrifuging and discarding the filtrate (90s, 4 deg.C, 12000rpm), placing the adsorption column back into the collection tube, repeating the step for 1 time, centrifuging at 4 deg.C and 12000rpm for 2min, removing the filtrate, opening the cover of the adsorption column, and standing for about 4min to completely remove ethanol residue on the adsorption column; (8) the dried columns were placed in standard RNase-Free 1.5mLEP tubes, 30. mu.L of deionized water was added dropwise to the center of each column, left at room temperature for 2min and centrifuged to collect the filtrate (2min, 4 ℃, 12000rpm), and the procedure was repeated 1 time to obtain more RNA solution.
The concentration (unit microgram/u) and purity of the extracted RNA are measured by an ultraviolet spectrophotometer, the concentration is adjusted to zero by DEPC water, and OD260/OD280 is between 1.8 and 2.0, which shows that the purity of the extracted RNA is good.
Reverse transcription into cDNA (reverse transcription as, rt): the reaction system is reverse transcription according to PrimeScript RT Master Mix kit of TaKaRa company: 5 PrimeScript RT Master MIX: 2 μ L, total RNA: 1 mu g of the solution; add ddH2O to the total volume of 10 mu L, placing the mixture into a PCR instrument for reverse transcription, and storing the reaction system at 37 ℃ for 15 minutes, 85 ℃ for 5 seconds and 4 ℃ for later use.
The real-time quantitative PCR method is characterized in that the expression of related genes is detected according to a SYBR Select Master Mix kit of appliedbiosystems (ACTB is an internal reference, ACTB-F: 5'-CTCGCCTTTGCCGATCC-3' is shown in SEQ ID NO.1, ACTB-R: 5'-TCTCCATGTCGTCCCAGTTG-3' is shown in SEQ ID NO.2, UGT2B 17-F: 5'-CCCTGGATCGAGCAGTCTTC-3' is shown in SEQ ID NO.3, UGT2B 17-R: 5'-GCTCAGTAACTTTTGTGTGGGG-3' is shown in SEQ ID NO.4, NKX 3.1-F: 5'-TGAGCAAGCAAGGGACTGAG-3' is shown in SEQ ID NO.5, NKX 3.1-R: 5'-TCTCCCCTCTACCAGCTCAC-3' is shown in SEQ ID NO.6, SLC45A 3-F: 5'-CCCAGGCTCAGGGTTAACAG-3' is shown in SEQ ID NO.7, SLC45A 3-R: 5'-GTGGTTAGGGAAGCCGTTGA-3' is shown in SEQ ID NO. 8): 17 μ L SYBR, 14 μ L RNase-free water, 1 μ L Forward primer, 1 μ L Reverse primer, 1 μ L LcDNA. The reaction procedure was Holding Stage: 2min at 50 ℃, 10min at 95 ℃, Cycling Stage at 95 ℃, 15s, 50 ℃, 1min and Number of Cycles at 40. CT values were calculated and the results presented as a histogram.
The results are shown in FIG. 3-B, where the interfering ARAP1 mRNA expression level was significantly reduced and all the downstream target gene mRNA levels of AR were significantly increased compared to the control group for the q-PCR detection cell line, indicating that ARAP1 inhibited AR-mediated gene transcription.
ARAP1 is a testis-specific antigen that is expressed only in normal testis and cancer: the applicant finds that the ARAP1 is only expressed in testis tissues in normal tissues by detecting mRNA expression of the ARAP1 in different tissues and corresponding cancers, and suggests that the ARAP1 is a novel testis-specific antigen (FIG. 1).
ARAP1 is a protein that interacts with the androgen receptor AR: the applicant carried out protein co-immunoprecipitation and GST-Pulldown experiments in breast cancer cells, and the results showed that ARAP1 can interact with androgen receptor AR with a direct region of action at the AR-carbon end. Immunofluorescence confocal experiments showed that the ARAP1 was mostly localized on the cytoplasm and cell membrane, and a few co-localized with AR in the nucleus (fig. 2).
ARAP1 down-regulates AR-mediated gene transcription: the luciferase double-gene report experiment shows that ARAP1 can obviously reduce the transcription activity of AR and inhibit the transcription of AR mediated genes; whereas qPCR experiments showed that silencing virus using ARAP1 could significantly up-regulate the cancer suppressor gene downstream of AR, suggesting that our silencing of ARAP1 could suppress cancer by up-regulating the cancer suppressor gene (fig. 3).
Transfection of siRNA knock-down ARAP1 following the cell transfection procedure in example 1 significantly reduced growth, invasion and metastasis of breast cancer cells: the cell growth curves show that silencing of ARAP1 significantly slowed the growth of breast cancer cells, while silencing of ARAP1 significantly promoted the invasion and metastasis of breast cancer cells (fig. 5).
Sequence listing
<110> university of Chinese medical science
<120> application of testis-specific expression protein ARAP1 in diagnosis and treatment of breast cancer
<141> 2021-10-26
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Claims (10)
1. The application of the reagent for detecting the expression level of the ARAP1 gene or the coded protein thereof in preparing a kit for diagnosing and screening breast cancer.
2. The use of the reagent for detecting the expression level of the ARAP1 gene or the protein coded by the same in the preparation of a kit for diagnosing and screening breast cancer according to claim 1, wherein the ARAP1 gene or the protein coded by the same is highly expressed in breast cancer cells.
3. The use of the reagent for detecting the expression level of the ARAP1 gene or the protein coded by the same in the preparation of a kit for diagnosing and screening breast cancer according to claim 1, wherein the use detects the expression level of the ARAP1 gene or the protein coded by the same in a sample by RT-PCR, real-time quantitative PCR, in situ hybridization, Northern blotting, a chip, a high-throughput sequencing platform, immunohistochemistry or enzyme-linked immunosorbent assay; the sample is tissue, serum or cells.
4. The use of the reagent for detecting the expression level of the ARAP1 gene or the protein coded by the same in the preparation of a kit for diagnosing and screening breast cancer according to claim 1, wherein the reagent contains a specific primer for amplifying the ARAP1 gene, a probe hybridized with the nucleotide sequence of the ARAP1 gene or an antibody specifically bound with the ARAP1 protein.
5. The use according to claim 4, wherein the specific primer sequences for amplifying the ARAP1 gene are as follows:
ARAP1-F：5’-AGAGTCCCAGGCAGGACAA-3’；
ARAP1-R：5’-GCAGAAGGCTGAGGAACACT-3’；
the antibody is a monoclonal antibody or a polyclonal antibody.
6. The use of any one of claims 1 to 5, wherein the breast cancer further comprises ER positive breast cancer.
Use of an inhibitor of expression of the ARAP1 gene in the manufacture of a medicament for the treatment of breast cancer.
8. The use of an inhibitor of the expression of the ARAP1 gene of claim 7, wherein the targeting sequence of the ARAP1 gene inhibitor is: 5'-TGGATTTGTACCATTCTTCT-3' are provided.
9. Use of an inhibitor of the expression of the ARAP1 gene according to claim 7 in the preparation of a medicament for the treatment of breast cancer, wherein the inhibitor comprises an siRNA or shRNA comprising the target sequence of claim 8.
10. Use of an inhibitor of the expression of the ARAP1 gene according to claim 7, in the manufacture of a medicament for the treatment of breast cancer, wherein the medicament comprises a pharmaceutically acceptable carrier and/or excipient.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160299143A1 (en) * | 2013-10-01 | 2016-10-13 | Ait Austrian Institute Of Technology Gmbh | Breast cancer diagnostic method and means |
| CN110592225A (en) * | 2019-11-05 | 2019-12-20 | 新乡医学院 | Triple negative breast cancer molecular marker and application thereof |
-
2021
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160299143A1 (en) * | 2013-10-01 | 2016-10-13 | Ait Austrian Institute Of Technology Gmbh | Breast cancer diagnostic method and means |
| CN110592225A (en) * | 2019-11-05 | 2019-12-20 | 新乡医学院 | Triple negative breast cancer molecular marker and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| SAMBAVY NADARAJA ET AL: "ARAP1 is an independent prognostic biomarker in older women with ovarian high-grade serous adenocarcinoma receiving first-line platinum-based antineoplastic therapy", MEDICAL ONCOLOGY, pages 40 - 47 * |
| SHIN-AE KANG ET AL: "PTK6 Inhibits Down-regulation of EGF Receptor through Phosphorylation of ARAP1", JBC, vol. 285, no. 34, pages 26013 - 26021 * |
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