Application of quercetin in preparation of glioblastoma ferroptosis inducer

Abstract

The invention provides application of quercetin in preparation of a glioblastoma ferroptosis inducer, and belongs to the technical field of biological medicine. Human-derived glioblastoma U251 cells are used for an in-vitro experiment, and it is proved that in glioblastoma, quercetin can reduce transcription of FTH1 genes and GPX4 genes, promote transcription of TFR1 genes, inhibit expression of FTH1 protein and GPX4 protein, reduce the content of GSH, improve the content of MDA and the content of Fe < 2 + >, promote ferroptosis of the human-derived glioblastoma U251 cells, and can be used for preparing a medicine for treating the glioblastoma. The growth and migration of the plants can be obviously inhibited. The invention further provides a glioblastoma ferroptosis inducer based on quercetin as an active component and application of the glioblastoma ferroptosis inducer in preparation of drugs for preventing and / or treating glioblastoma.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of quercetin in the preparation of a glioblastoma ferroptosis inducer. Background Art

[0002] Glioblastoma Multiforme (GBM) is a highly malignant tumor of the central nervous system, characterized by strong invasiveness, high recurrence rate, and extremely poor prognosis. Although standard treatment options (surgical resection combined with temozolomide chemotherapy and radiotherapy) can temporarily alleviate the disease, the median survival of patients is usually only about 14 months, and the 5-year survival rate is less than 6%. In addition, the current standard treatment for glioblastoma, temozolomide (TMZ), still has problems such as drug resistance (GBM cells repair DNA damage through MGMT overexpression, resulting in reduced efficacy of TMZ), toxic side effects (TMZ causes hematological toxicity, neutropenia, anemia, etc., which limits long-term use), and insufficient targeting (lack of specific killing of tumor stem cells, prone to recurrence). Therefore, the continued research and development of glioblastoma-related drugs remains a key issue in this field.

[0003] Ferroptosis, an iron-dependent, lipid peroxidation-driven, non-apoptotic cell death, has become a research hotspot in cancer treatment in recent years. It depletes glutathione (GSH) and inhibits glutathione peroxidase 4 (GPX4) activity, triggering lipid peroxidation accumulation and bypassing traditional apoptotic resistance mechanisms. However, existing ferroptosis inducers (such as erastin and RSL3) have limitations such as poor targeting (lack of specificity for tumor cells and prone to damage to normal tissues), synthetic drug toxicity (some small molecule compounds are difficult to clinically translate due to systemic toxicity), and a single mechanism (reliance on a single target and easily offset by compensatory pathways), making them difficult to use for the targeted treatment of glioblastoma. Therefore, exploring ferroptosis inducers targeting glioblastoma may be the key to solving the problem of glioblastoma-related drug development. Summary of the Invention

[0004] In order to solve the problems existing in the prior art, the object of the present invention is to provide the use of quercetin in the preparation of a glioblastoma ferroptosis inducer.

[0005] The present invention also aims to provide a glioblastoma ferroptosis inducer.

[0006] The present invention also aims to provide the use of the above-mentioned glioblastoma ferroptosis inducer in the preparation of a drug for preventing and / or treating glioblastoma.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides the use of quercetin in preparing a glioblastoma ferroptosis inducer.

[0009] Preferably, the quercetin reduces the transcription of FTH1 gene and GPX4 gene in glioblastoma.

[0010] Preferably, the quercetin promotes the transcription of TFR1 gene in glioblastoma.

[0011] Preferably, the quercetin inhibits the expression of FTH1 protein and GPX4 protein in glioblastoma.

[0012] Preferably, the quercetin reduces the GSH content in glioblastoma.

[0013] Preferably, the quercetin increases the MDA content and Fe 2+ content.

[0014] Preferably, the quercetin inhibits glioblastoma cell proliferation.

[0015] Preferably, the quercetin inhibits glioblastoma cell migration.

[0016] The present invention also provides a glioblastoma ferroptosis inducer, wherein the glioblastoma ferroptosis inducer uses quercetin as an active ingredient, and the content of the quercetin is 50-100 wt%.

[0017] The present invention also provides use of the glioblastoma ferroptosis inducer in the preparation of a medicament for preventing and / or treating glioblastoma.

[0018] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0019] This study proposes the use of quercetin in the preparation of a glioblastoma ferroptosis inducer. In vitro experiments using human glioblastoma U251 cells demonstrated that quercetin can promote ferroptosis in U251 cells and significantly inhibit their growth and migration. Furthermore, quercetin bypasses the TMZ resistance mechanism through a ferroptosis pathway (not DNA damage), without significant side effects. It can regulate multiple targets, simultaneously inhibiting proliferation and migration and inducing ferroptosis. This approach is superior to traditional drugs with a single mechanism and, compared to other ferroptosis inducers, offers higher efficacy and lower cytotoxicity. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 : Inhibitory rate of quercetin on U251 cells;

[0021] Figure 2 :Effect of quercetin on the cloning ability of U251 cells;

[0022] Figure 3 :Effects of quercetin on the migration of U251 cells;

[0023] Figure 4 :Effects of quercetin on ferroptosis-related genes;

[0024] Figure 5 :Effects of quercetin on ferroptosis-related proteins;

[0025] Figure 6 :Quercetin on Fe 2+ , MDA and GSH contents. DETAILED DESCRIPTION

[0026] The present invention provides the use of quercetin in the preparation of a glioblastoma ferroptosis inducer. In vitro experiments were conducted on human glioblastoma U251 cells. The results showed that quercetin can reduce the transcription of FTH1 gene and GPX4 gene in glioblastoma, promote the transcription of TFR1 gene in glioblastoma, inhibit the expression of FTH1 protein and GPX4 protein in glioblastoma, reduce the GSH content of glioblastoma, and increase the MDA content and Fe content of glioblastoma. 2+ It was demonstrated that quercetin can promote the occurrence of ferroptosis in U251 cells, and quercetin can be used as a ferroptosis inducer for glioblastoma cells or for the preparation of a ferroptosis inducer for glioblastoma cells.

[0027] The present invention detects the cell viability, proliferation ability, and migration ability of human glioblastoma U251 cells, confirms that quercetin significantly inhibits glioblastoma cell proliferation and cell migration by inducing ferroptosis in U251 cells, and verifies the proliferation inhibitory effect of quercetin on U251 cells and the regulation of tumor invasion and metastasis characteristics.

[0028] The present invention also provides a glioblastoma ferroptosis inducer, wherein the glioblastoma ferroptosis inducer comprises quercetin as an active ingredient, and the content of the quercetin is 50-100 wt %. The glioblastoma ferroptosis inducer of the present invention can comprise quercetin as the sole active ingredient, or quercetin can be used in combination with other active ingredients.

[0029] The present invention also provides the use of the above-mentioned glioblastoma ferroptosis inducer in the preparation of a drug for preventing and / or treating glioblastoma. The glioblastoma ferroptosis inducer can be used as a drug for treating glioblastoma, and can also be used in combination with other active ingredients to prepare a drug for preventing and / or treating glioblastoma.

[0030] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0031] In the specific embodiments of the present invention, quercetin was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; DMEM / F12 complete medium was purchased from GIBCO, USA, containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin double antibody; CCK-8 kit and crystal violet staining solution were purchased from Beijing Beiren Chemical Technology Co., Ltd. and diluted according to the instructions; MDA and GSH detection kits were purchased from Abcam, UK, and operated according to the instructions.

[0032] In the specific examples of the present invention, all experiments were performed with three biological replicates, and the data were expressed as mean ± standard deviation. Statistical analysis was performed using GraphPad Prism 9.0 (*P < 0.05, **P < 0.01, ***P < 0.001).

[0033] In the following examples, unless otherwise specified, all methods are conventional.

[0034] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0035] Example 1

[0036] 1. Functional phenotypic testing

[0037] Cell line: human glioblastoma U251 cells.

[0038] Culture conditions: 37°C, 5% CO2, DMEM / F12 complete medium for adherent culture, medium change every 3 days.

[0039] Experimental groups: U251 cell group and drug treatment group; the cells in the U251 cell group were cultured normally, and the cells in the drug treatment group were treated with culture medium containing different concentrations of quercetin (quercetin was dissolved in DMSO and then diluted to DMEM / F12 complete medium) for 48 hours.

[0040] (1) Cell viability assay

[0041] CCK-8 assay: U251 cells in the logarithmic growth phase were washed twice with PBS buffer and digested with 0.1% trypsin. After digestion, the cells were centrifuged and the precipitate was collected. DMEM / F12 complete medium was added to make a single cell suspension. The cells were counted on a hemocytometer and different treatment groups were prepared. The cell number was 2×10 5After mixing, the cells were seeded into a 96-well plate at a volume of 100 μL per well, with 3 replicates per group. After seeding, the cells were cultured in an incubator.

[0042] After the cells have grown normally, the old culture medium was replaced with 90 μL of culture medium containing different drug concentrations (quercetin concentrations of 0 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, and 90 μM) at 48 hours. 10 μL of CCK-8 reagent was added to each well at the corresponding time. Incubate in the dark for 4 hours. Before detection, gently shake the 96-well plate to mix the culture medium in the wells. The absorbance value (450 nm) of the test wells was measured with a microplate reader and the results were recorded. The experiment was repeated three times, and Graphpad Prism 9.0 software was used for plotting and statistical analysis.

[0043] The inhibition rate of quercetin on U251 cells is as follows Figure 1 As shown in the figure, A: U251 cell viability under different quercetin concentrations; B: Quercetin inhibition curve of U251 cell viability. CCK-8 results showed that quercetin had a significant concentration-dependent inhibition on the proliferation of U251 cells (A). Nonlinear regression analysis calculated that the half-maximal inhibitory concentration (IC50) of quercetin on U251 cells treated for 48 hours was 50 ) was 46.95 μM (B).

[0044] (2) Proliferation inhibition assay

[0045] Plate colony formation assay: U251 cells were washed twice with PBS buffer, digested, centrifuged, and resuspended. Counted using a hemocytometer and 1×10 4 Cells were inoculated into 2 mL of culture medium containing different drug concentrations (35 μM and 70 μM quercetin, respectively). Cultured in an incubator, the medium was changed every 5 days. After 14 days, the cells were observed and photographed. A cell count of ≥50 was considered the cloning standard.

[0046] Before imaging, cells were stained by removing the culture medium and rinsing twice with PBS. Fix the cells with 4% paraformaldehyde for 30 minutes at room temperature, then rinse and aspirate any remaining PBS. Stain the cells with 0.1% crystal violet solution for 20 minutes and rinse with water to remove the violet. Images were taken and analyzed using Image J software to count colonies. The experiment was repeated three times, and graphing and statistical analysis were performed using Graphpadprism 9.0 software.

[0047] Effect of quercetin on the cloning ability of U251 cells Figure 2As shown in the figure, Con represents the U251 cell group (blank control group), 35μM represents the drug-treated group with a quercetin concentration of 35μM, and 70μM represents the drug-treated group with a quercetin concentration of 70μM. Quantitative analysis results showed that the number of colonies formed in the blank control group reached 193.59±4.66, while the number of colonies formed after treatment with 35μM quercetin was 89.68±7.41, with an inhibition rate of 53.68%. When the quercetin concentration was increased to 70μM, the number of colonies further decreased to 15.46±3.79, with an inhibition efficiency of 92.01%. Statistical analysis showed that quercetin's inhibitory effect on U251 cell proliferation was significantly concentration-dependent.

[0048] (3) Migration inhibition assay

[0049] Scratch assay: U251 cells in the logarithmic growth phase were washed twice with PBS buffer and digested with trypsin. After digestion, the cells were collected by centrifugation. DMEM / F12 complete medium was added and gently pipetted to form a single-cell suspension. The cells were counted on a hemocytometer and seeded into a six-well plate with a pre-marked bottom. The seeding number of U251 cells was 5.0×10 5 Each well was filled with culture medium containing different drug concentrations (quercetin concentrations of 35 μM and 70 μM, respectively) to a volume of 2 mL, and the cells were placed in a 37°C, 5% CO2 incubator in the dark.

[0050] When the cells are growing well and the cell density is high, use a 200μL pipette tip to scratch. After rinsing and absorbing the remaining PBS, add 2mL of culture medium with different concentrations of drugs (quercetin concentrations are 35μM and 70μM, respectively). Take pictures under a microscope and record the photos of each concentration treatment group at 0h. After continuing to culture for 48h, rinse and absorb the remaining PBS, add basal culture medium, and take pictures again. Use Image J software to analyze the scratch surface and calculate the cell migration rate. Repeat the experiment 3 times and use Graphpadprism 9.0 software to draw statistical graphs.

[0051] Effects of quercetin on the migration of U251 cells Figure 3 As shown in the figure, Con represents the U251 cell group (blank control group), 35μM represents the drug-treated group with a quercetin concentration of 35μM, and 70μM represents the drug-treated group with a quercetin concentration of 70μM. The results showed that the cells in the blank control group had a very strong scratch healing ability after 48 hours, with a migration rate of 24.38±4.01%, showing typical invasive growth characteristics. After treatment with 35μM quercetin, the migration rate dropped to 15.71±1.25%, and the inhibition efficiency reached 35.56%; when the quercetin concentration was increased to 70μM, the migration rate further decreased to 4.34±1.64%, and the inhibition efficiency was as high as 82.20%.

[0052] 2. Molecular mechanism verification

[0053] Cell line: human glioblastoma U251 cells.

[0054] Culture conditions: 37°C, 5% CO2, DMEM / F12 complete medium for adherent culture, medium change every 3 days.

[0055] Experimental groups: U251 cell group and drug treatment group; the cells in the U251 cell group were cultured normally, recorded as Con; the drug treatment groups were treated with culture medium with quercetin concentrations of 35 μM and 70 μM (quercetin was dissolved in DMSO and diluted to DMEM / F12 complete medium) for 48 hours, recorded as 35 μM and 70 μM, respectively.

[0056] (1) Gene expression detection

[0057] RT-qPCR: Total cellular RNA was extracted using the Trizol method: U251 cells from different groups were digested and centrifuged, and the supernatant was discarded. 500 μL of Trizol lysis buffer was added and allowed to stand for 5 minutes. 100 μL of pre-chilled chloroform was added, and the tube was shaken manually for 15 seconds. The tube was allowed to stand for 5 minutes, and then centrifuged. The supernatant was aspirated and an equal volume of pre-chilled isopropanol was added. Mix thoroughly, and the tube was allowed to stand at 25°C for 10 minutes before centrifugation to precipitate RNA. The supernatant was discarded, and 0.5 mL of pre-chilled ethanol was added. The tube was gently inverted to wash the tube walls and centrifuged. After centrifugation, the ethanol was aspirated and the pellet was retained (repeat this wash twice). Dry at room temperature for 3 minutes. Dissolve the supernatant in 10 μL of DEPC water.

[0058] Prepare cDNA by reverse transcription. First, prepare system 1 (Table 1) (purchased from Invitrogen, USA). After reacting at 65°C for 5 minutes, immediately place it on ice to cool for 2 minutes. Prepare system 2 (Table 2), add it to system 1 to prepare a 20μL system, mix and centrifuge. Set the reaction program: 42°C for 40 minutes, 72°C for 10 minutes to obtain the cDNA product. Use the cDNA product as a template (primer sequence see Table 3) (purchased from Shanghai Shenggong Biological Co., Ltd.) and prepare the following reaction system 3 (Table 4) (purchased from Takara, Japan) to carry out the PCR process. The reaction program is: 95°C for 4 minutes, 95°C for 30 seconds, 60°C for 30 seconds, 72°C for 30 seconds, and 40 cycles. Obtain the Ct value for each reaction, using GAPDH as the internal reference according to the relative expression ratio = 2 -ΔΔCt The formula is used to process the experimental data of each group to obtain the relative expression value of the target gene.

[0059] Table 1 Reverse transcription reaction system 1

[0060] Reagent name Add volume Total RNA 2.0 μg 50μM Oligo DT Primer 1.0 μL 10mM dNTP 1.0μL DEPC water Make up to 10.0 μL

[0061] Table 2 Reverse transcription reaction system 2

[0062] Reagent name Add volume 5×Prime Script Ⅱ Buffer 4.0μL 200U / μL Prime Script II RTase 1.0μL 40U / μL RNase inhibitor 0.5μL DEPC water 4.50 μL

[0063] Table 3 Primer sequences used in PCR reaction

[0064]

[0065] Table 4 RT-qPCR reaction system 3

[0066] Reagent name Add volume 2×SYBR Premix Ex Taq 10.0μL Upstream primer 0.8μL Downstream primer 0.8μL 5U / μL Taq DNA polymerase 0.2μL cDNA products 2.0 μL DEPC water 6.2μL

[0067] Effects of quercetin on ferroptosis-related genes Figure 4 The results showed that at the mRNA level, quercetin significantly reduced the transcription of FTH1 and GPX4, while promoting the transcription of TFR1. The GPX4 gene decreased by 31.48% and 57.88%, the FTH1 gene decreased by 37.86% and 67.39%, and the TFR1 gene increased by 123.65% and 65.49%, respectively.

[0068] (2) Protein expression detection

[0069] To extract cellular proteins: Remove quercetin-treated cells, discard the culture medium, digest and centrifuge, resuspend in pre-chilled PBS, transfer to a 1.5 mL Eppendorf tube, and wash the supernatant. Add 3 μL of PMSF protease inhibitor and 300 μL of RIPA lysis buffer to each tube. Shake on ice for 30 minutes, beat to mix, and centrifuge at 12,000 rpm for 15 minutes at 4°C. Pipette the supernatant into a pre-chilled 600 μL Eppendorf tube and store at -20°C until needed.

[0070] To determine protein concentration using the BCA assay, prepare the colorimetric working solution: Reagents A and B (50:1) and mix thoroughly. Add the standard to a 96-well plate. Next, add the protein sample to be tested, along with the diluted sample. Add 200 μL of the colorimetric working solution to each well and incubate at 37°C in the dark for 30 minutes. Measure the absorbance of each well (560 nm) using a microplate reader. The protein concentration is then directly determined using the reader's pre-programmed protocol.

[0071] Western blot: Set the sample load to 10 μg per protein sample. Calculate the required volume based on the respective concentrations, add RIPA buffer to a volume of 20 μL, add 5 μL of 5× instant loading buffer, and denature in a metal bath at 100°C for 5 minutes. Cool on ice for 2 minutes, centrifuge at high speed for 5 minutes, and use the supernatant for electrophoresis analysis. Mix 5 mL of lower gel solution, 5 mL of lower gel buffer, and 50 μL of coagulant, then add to a PAGE gel plate. Seal with purified water. Once the lower gel has solidified, discard the upper water layer. Add 3 mL of upper gel solution, 3 mL of upper gel buffer, and 30 μL of coagulant to the PAGE gel plate. Select a 15-tooth comb. Once prepared, remove the comb and blow through with a syringe to remove air bubbles and impurities.

[0072] Add sample and the dual-color pre-stained Protein Maker according to the volume added to each well. Turn on the power supply for electrophoresis. When the protein is in the stacking gel, maintain a voltage of 90V and a current of 40mA for approximately 30 minutes. Once in the separating gel, maintain a voltage of 120V and a current of 40mA for approximately 40 minutes before ending the electrophoresis. After electrophoresis, select the target protein band for segmentation. Soak the resulting gel in TBST for 2 minutes. Equilibrate the gel and PVDF membrane in transfer buffer for approximately 10 minutes. Place the sponge pad, gel, PVDF membrane, and sponge pad in order (be careful to avoid trapping bubbles). Clamp the membrane and transfer it. Maintain a voltage of 120V and a current of 40mA for 1 hour. Afterwards, cut the PVDF membrane according to the molecular weight of the target protein, mark it, and soak it in TBST for 5 minutes. Add the prepared blocking solution to the membrane and gently shake it on a shaker. Block it at room temperature for approximately 30 minutes. Rinse it with TBST three times, each 5 minutes.

[0073] The membrane was placed in the primary antibody dilution buffer containing the following antibodies: GPX4 (Catalog No: 67763-1-Ig, 1:4000), FTH1 (Catalog No: 83428-1-R, 1:5000), TFR1 (Catalog No: 66180-1-Ig, 1:5000), and β-Actin (Catalog No: 66009-1-Ig, 1:20,000). The membrane was incubated overnight at 4°C, followed by three rinses in TBST. The membrane was then added to the corresponding secondary antibody dilution buffer, incubated on a shaker at room temperature for 1 hour, and then rinsed three times in TBST. The membrane was removed from the wash buffer, drained, and kept moist. An equal volume of chemiluminescent solution was mixed and evenly applied to the membrane. The reaction was allowed to proceed for 2 minutes. The membrane, with the working solution, was then placed on a shaker. The membrane was removed, drained, and directly photographed using an imaging device. The images were saved and analyzed. The images were analyzed using Image J software, and the intensity-area sum was calculated. The grayscale value of the control group was set to 1, and the relative expression of the grayscale value of the experimental group was calculated based on this value (the antibody was purchased from Wuhan Tri-Eagle Biotechnology Co., Ltd.).

[0074] Effects of quercetin on ferroptosis-related proteins Figure 5 As shown in the figure, Western blot analysis showed that quercetin significantly inhibited the expression of GPX4 protein. In the 35μM and 70μM quercetin treatment groups, GPX4 protein expression decreased by 20.62% and 53.28%, respectively. In addition, FTH1 protein expression was also significantly reduced by 48.09% and 61.91%, respectively. TFR1 protein expression was significantly increased in the 70μM quercetin treatment group.

[0075] (3) Detection of ferroptosis markers

[0076] MDA assay: First, lyse U251 cells in an ice bath, and measure protein concentration using a BCA assay kit. Prepare the standard and working solution according to the manufacturer's instructions. Add the control solution, standard, and sample to a centrifuge tube, followed by the working solution. Mix thoroughly, then heat at 100°C for 15 minutes. Cool to room temperature, centrifuge, and transfer the supernatant to a 96-well plate. Measure the absorbance of each well (532 nm) using a microplate reader to calculate the MDA concentration.

[0077] GSH assay: First, disrupt cells by ultrasonication. Add a precipitant to the cell suspension, mix thoroughly, centrifuge, and collect the supernatant for color development. Add the treated standard and sample to a 96-well plate, add the color development reagent and buffer, mix thoroughly, and let stand for 5 minutes. Measure the absorbance of each well (405 nm) using a microplate reader to calculate the GSH concentration.

[0078] Fe 2+ Detection: Dilute 100mM iron standard solution with distilled water to generate 0, 2, 4, 6, 8, and 10nM standards.5 After digestion and centrifugation of each U251 cell, add 100 μL of iron assay buffer and quickly homogenize, centrifuge at 4°C, 16000 rpm for 10 minutes, and take the supernatant for subsequent experiments. Add 50 μL of sample to the sample well of the 96-well plate, and add iron assay buffer to make the volume of each well 100 μL. Add 5 μL of iron assay buffer to each sample well. Mix evenly using a horizontal shaker or pipette, and incubate the reaction at 25°C in the dark for 30 minutes. Add 100 μL of iron probe to each standard and sample well, mix evenly using a horizontal shaker or pipette, and incubate the reaction at 25°C in the dark for 60 minutes. Measure the absorbance of each well (593 nm) with a microplate reader and calculate Fe 2+ concentration (the kit was purchased from Abcam, UK).

[0079] Quercetin on Fe 2+ , MDA and GSH content as Figure 6 The results showed that compared with the blank control group, 35 μM and 70 μM quercetin reduced GSH by 16.04% and 30.36% in U251 cells, respectively, while MDA content increased by 67.25% and 98.13%, respectively. 2+ The contents of quercetin and ferroptosis increased by 32.71% and 84.44% respectively. The results showed that quercetin can promote the occurrence of ferroptosis.

[0080] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. Application of quercetin in the preparation of glioblastoma ferroptosis inducer.

2. The use according to claim 1, characterized in that The quercetin reduces the transcription of the FTH1 gene and the GPX4 gene in glioblastoma.

3. The use according to claim 1, characterized in that The quercetin promotes the transcription of the TFR1 gene in glioblastoma.

4. The use according to claim 1, characterized in that The quercetin inhibits the expression of FTH1 protein and GPX4 protein in glioblastoma.

5. The use according to claim 1, characterized in that The quercetin reduces the GSH content in glioblastoma cells.

6. The use according to claim 1, characterized in that The quercetin increases the MDA content and Fe 2+ content.

7. The use according to claim 1, characterized in that The quercetin inhibits glioblastoma cell proliferation.

8. The use according to claim 1, characterized in that The quercetin inhibits glioblastoma cell migration.

9. A glioblastoma ferroptosis inducer, characterized in that: The glioblastoma ferroptosis inducer uses quercetin as an active ingredient, and the content of the quercetin is 50-100 wt %.

10. Use of the glioblastoma ferroptosis inducer according to claim 9 in the preparation of a medicament for preventing and / or treating glioblastoma.

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