Application of mitoxantrone in the preparation of drugs for treating head and neck squamous cell carcinoma with high IL6R expression

Abstract

The present invention discloses the use of mitoxantrone in the preparation of a drug for treating head and neck squamous cell carcinoma with high IL6R expression. The present invention discovers for the first time that IL6R can be used as a biomarker for mitoxantrone in the treatment of head and neck squamous cell carcinoma. The higher the expression level of IL6R, the more sensitive the head and neck squamous cell carcinoma cells are to mitoxantrone. Based on this, the present invention provides a new use of mitoxantrone: mitoxantrone can be used to prepare a drug for treating head and neck squamous cell carcinoma with high IL6R expression. At the same time, the present invention also provides a kit for detecting the sensitivity of head and neck squamous cell carcinoma patients to mitoxantrone. The kit can pre-detect whether the head and neck squamous cell carcinoma patient is suitable for treatment with mitoxantrone before treatment, so that personalized treatment plans can be formulated for different patients, improving the treatment effect and the patient's quality of life.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the application of mitoxantrone in preparing a medicine for treating head and neck squamous cell carcinoma. Background Art

[0002] Head and neck cancer is the sixth most common type of cancer in the world. It includes epithelial malignancies originating in the paranasal sinuses, nasal cavity, oral cavity, pharynx, and larynx. Globally, it causes more than 550,000 cases and more than 300,000 deaths each year. More than 95% of head and neck cancers are head and neck squamous cell carcinoma (HNSCC). Currently, surgery, radiotherapy, and chemotherapy remain the main treatments for HNSCC, but these treatments can cause damage to the organs and functions of the head and neck, leading to a decrease in patients' quality of life. In recent years, with the advancement of sequencing and multi-omics analysis technologies, targeted therapies have demonstrated high efficiency and low toxicity, becoming the mainstream treatment for many types of cancer. However, because HNSCC is mainly caused by mutations in tumor suppressor genes and there are fewer oncogenic driver genes available for intervention, targeted drug treatments are still extremely scarce.

[0003] Mitoxantrone (CAS: 70476-82-3) is a long-established anthracycline compound with anti-tumor properties. Because it lacks an amino sugar structure, does not generate free radicals, and inhibits lipid peroxidation, it exhibits low cardiotoxicity. Mitoxantrone's anti-tumor activity is comparable to or slightly higher than that of doxorubicin, and significantly higher than that of cytarabine, cyclophosphamide, and fluorouracil. It is primarily used to treat acute myeloid leukemia, malignant lymphoma, and breast cancer, as well as various other cancer types, including bladder cancer, ovarian cancer, gastrointestinal tumors, malignant mesothelioma, and multiple sclerosis. However, mitoxantrone has a low drug response rate in head and neck squamous cell carcinoma (HNSCC), and thus has not been clinically used in this setting. Summary of the Invention

[0004] The present invention aims to provide the use of mitoxantrone in preparing a medicament for treating head and neck squamous cell carcinoma with high IL6R expression.

[0005] In order to achieve the above object, the present invention provides a new use of mitoxantrone: mitoxantrone can be used to prepare a drug for treating head and neck squamous cell carcinoma with high IL6R expression.

[0006] Optionally, the above-mentioned drug for treating head and neck squamous cell carcinoma with high IL6R expression comprises an IL6R agonist.

[0007] The present invention also provides a pharmaceutical composition for preparing a treatment of head and neck squamous cell carcinoma, wherein the pharmaceutical composition comprises at least an IL6R agonist and mitoxantrone.

[0008] The present invention also provides a kit for detecting the sensitivity of head and neck squamous cell carcinoma patients to mitoxantrone, and the kit at least comprises a reagent for detecting the expression level of IL6R.

[0009] Optionally, the kit detects the expression level of IL6R in head and neck squamous cell carcinoma tissues by RT-PCR.

[0010] Optionally, the above reagents include a nucleic acid that binds to the IL6R gene, and the nucleic acid may be a primer for amplifying the IL6R gene.

[0011] Optionally, the kit detects the expression level of IL6R in head and neck squamous cell carcinoma tissues by immunohistochemistry.

[0012] Optionally, the above reagent includes a substance that binds to the IL6R protein, and the substance may be an antibody that specifically binds to the IL6R protein.

[0013] Compared with the prior art, the technical effects of the present invention are as follows:

[0014] The present invention discovers for the first time that IL6R can serve as a biomarker for mitoxantrone in the treatment of head and neck squamous cell carcinoma (HNSCC). The higher the expression of IL6R, the more sensitive the HNSCC cells are to mitoxantrone. Based on this, the present invention provides a new use for mitoxantrone: mitoxantrone can be used to prepare a drug for treating HNSCC with high IL6R expression. Furthermore, the present invention provides a kit for detecting the sensitivity of HNSCC patients to mitoxantrone. This kit can be used to pre-test whether HNSCC patients are suitable for mitoxantrone treatment before treatment, thereby enabling the development of personalized treatment plans for different patients, improving treatment efficacy and quality of life. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This figure shows the results of high-throughput drug screening of 13 primary tumor cells using 2248 drugs in the present invention.

[0016] Figure 2 This is a diagram showing the results of high-throughput drug screening of 300 drugs among the 2248 drugs of the present invention that can induce more than 50% cell killing effect in a primary tumor cell line.

[0017] Figure 3 This figure shows the results of the efficacy test of mitoxantrone on 10 primary tumor cells.

[0018] Figure 4 This is the expression result of the top 7 genes in the pharmacogenomics analysis between the mitoxantrone-sensitive and mitoxantrone-resistant cell groups.

[0019] Figure 5 The graph shows the expression level of IL6R in primary tumor cells PDC_52 and commercial cell lines HSC3 and TU686 after knocking down IL6R expression using small interfering RNA. Figure 5A is the expression level result of IL6R in PDC_52 after small interfering RNA knocked down the expression of IL6R in PDC_52. Figure 5 B is the expression level of IL6R in HSC3 after small interfering RNA knocked down the expression of IL6R in HSC3. Figure 5 C is the expression level of IL6R in TU686 after small interfering RNA knocked down the expression of IL6R in TU686.

[0020] Figure 6 The relative cell survival rates of cells in each group after IL6R knockdown in PDC_52, HSC3 and TU686 were treated with 0.5 μM mitoxantrone for 72 h. Figure 6 A is the relative cell survival rate of IL6R knockdown PDC_52 cells after treatment with 0.5 μM mitoxantrone for 72 hours. Figure 6 B is the relative cell survival rate of IL6R knockdown HSC3 cells after treatment with 0.5 μM mitoxantrone for 72 h. Figure 6 C is the relative cell survival rate of TU686 cells with IL6R knockdown after treatment with 0.5 μM mitoxantrone for 72 h.

[0021] Figure 7 The results of PDX_14 transplanted tumors treated with mitoxantrone drug and solvent control were shown. Figure 7 A is a statistical graph of PDX_14 treated with mitoxantrone drug and PDX_14 treated with solvent control. Figure 7 B is a photo of the transplanted tumors of PDX_14 treated with mitoxantrone and PDX_14 treated with solvent as a control.

[0022] Figure 8 The results of PDX_31 transplanted tumors treated with mitoxantrone drug and solvent control were shown. Figure 7 A is a statistical graph of PDX_31 treated with mitoxantrone drug and PDX_31 treated with solvent control. Figure 7 B is a photo of the transplanted tumors of PDX_31 treated with mitoxantrone and PDX_31 treated with solvent as a control. DETAILED DESCRIPTION

[0023] The present invention is described in detail below by way of examples. It is necessary to point out that the following examples are only used to further illustrate the present invention and are not to be construed as limiting the scope of protection of the present invention. Those skilled in the art may make some non-essential improvements and adjustments to the present invention based on the above-mentioned disclosure. In the following examples, unless otherwise indicated, the reagents used are all analytically pure, and the reagents and commercial cell lines used are all available from commercial channels. Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention.

[0024] IL6R (Interleukin-6 receptor) is an 80kDa glycoprotein also known as gp80 and CD126. IL6R binds to IL-6 through residues within domains 2 and 3, recruiting gp130 to form a high-affinity IL6 / IL6R / gp130 complex capable of signal transduction, leading to activation of downstream signaling. Studies have shown that abnormal activation of the IL6 / IL6R pathway is associated with poor prognosis in various cancers, including colorectal cancer, liver cancer, and breast cancer.

[0025] The inventors of the present invention selected 2,248 drugs (including drugs approved by the U.S. Food and Drug Administration, drugs evaluated in clinical trials, and preclinical candidate compounds) and conducted three rounds of high-throughput drug screening on patient-derived cell (PDC) head and neck squamous cell carcinoma cells derived from patient tumors and commercial head and neck squamous cell carcinoma cell lines. They finally found that mitoxantrone has a killing effect on head and neck squamous cell carcinoma cells with high IL6R expression, and the higher the expression level of IL6R, the better the killing effect of mitoxantrone on head and neck squamous cell carcinoma cells.

[0026] Example 1

[0027] A patient-derived xenograft (PDX) model of head and neck squamous cell carcinoma was constructed for subsequent in vivo transplantation tumor validation experiments.

[0028] For patients whose PDX models are constructed using this method, clinical information is recorded, including basic information (gender, age, smoking and alcohol history, etc.), clinical pathological diagnosis (tumor size and location, TNM stage, HPV infection status), previous treatment history (surgery, radiotherapy), and prognostic information such as recurrence and metastasis. Tumor, adjacent tumor, and blood samples are collected from the patients. Tumor samples are then subjected to histopathological histomorphological and genetic analysis.

[0029] After the tumor tissue is surgically removed, observe the color, morphology, and texture of the tissue, remove the necrotic tissue, and select the central part of the lesion for sampling. Because head and neck squamous cell carcinoma generally grows in contaminated areas such as the oral cavity and nasal mucosa, the sample needs to be sterilized with 0.05% sodium hypochlorite and quickly washed with 1% penicillin-streptomycin double-antibody PBS for 30 seconds before inoculation and transplantation into mice. Gently scrape the peripheral tissue of the tissue sample and cut the tumor into 1-2 mm pieces. 3 Small pieces of the patient's tissue are transplanted under sterile conditions into the blood supply and lymph node-rich areas (such as the bilateral axilla) of immunodeficient mice to construct a subcutaneous PDX model, or inoculated into the bilateral submandibular space of the animal to construct an in situ PDX model. To increase the success rate of inoculation, Matrigel can be mixed with the patient's tumor tissue and then inoculated. Each patient tissue can be inoculated into 3-5 mice. The growth trajectory of the PDX model begins to be tracked 1-2 weeks after modeling. When the tumor volume exceeds 800mm 3 Or when there is no significant increase in tumor volume for two weeks, the transplanted tumor is serially passaged. Generally, when the transplanted tumor is passaged for more than three generations, the model is considered to be stable for passage, and each model is named PDX_N (N is an integer greater than zero).

[0030] Example 2

[0031] Patient-derived cells (PDCs) from head and neck squamous cell carcinoma patients were constructed for subsequent in vitro validation experiments.

[0032] After transporting head and neck squamous cell carcinoma tumor tissue back to the laboratory at 4°C, the outer packaging was disinfected and the tissue was transferred to the cell room through a sterile transfer chamber. Sterile microscissors and microtweezers were used to dissect and mince the tissue. A tissue digestion solution (1 mg / ml collagenase type IV, 200 U / ml hyaluronidase, 200 U / ml DNase type I, and 1× phenol red-free trypsin) was prepared, with the volume of the tissue digestion solution being approximately 5-10 times the tumor volume.

[0033] After centrifugation at 1500 rpm for 3 minutes, resuspend the minced tissue in tissue digestion buffer, transfer it to a tissue dissociation tube, and place it in a Miltenyi tissue processor. Select a separation mode based on tissue toughness. After mincing, digest the tissue on a shaker at 37°C for 30-60 minutes. Shake the tube up and down every 10 minutes to mix the tissue and prevent clumping, which can affect digestion. Digestion is terminated when the digestion buffer becomes turbid and no tissue remains. Neutralize the digestion with culture medium supplemented with 10% fetal bovine serum and pass the tube through 100μm and 40μm filters, collecting any undigested clumps and collecting the cell suspension. After centrifugation at 1500 rpm for 3 minutes, if significant erythrocyte contamination is detected, add 1 ml of erythrocyte lysis buffer. After 3 minutes of rest, dilute with 10 ml of PBS and centrifuge at 1500 rpm for 3 minutes.

[0034] Discard the supernatant and resuspend in 5 ml of complete medium. The complete medium formula is high-glucose DMEM medium: DMEM / F12 medium = 3:1; insulin, 5 μg / ml; amphotericin B, 250 ng / ml; gentamicin, 10 μg / ml; cholera toxin, 0.1 nM; EGF, 0.125 ng / ml; hydrocortisone, 25 ng / ml; ROCK inhibitor Y-27632, 10 μM. Sterilize the solution with a 0.22 μm sterile filter and store at 4°C for 2 months. Incubate the cell suspension in a humidified incubator at 37°C, 5% CO2 for 2 months. PDC cells that maintain high cell proliferation activity after 5 or more successful passages are considered successfully constructed, and each PDC cell line is designated PDC_N (N is an integer greater than zero).

[0035] Example 3 High-throughput drug screening for the purpose of "new uses of old drugs" using primary head and neck squamous cell carcinoma cells and cell lines

[0036] Given the high cost, long development cycle, and extremely high failure rate of new drug development, repurposing drugs with known indications to treat other diseases is becoming an increasingly promising approach to drug development, commonly known as "repurposing old drugs." Compared to developing completely new drugs, this approach offers the following advantages: 1. A higher success rate, as marketed drugs have already undergone systematic safety evaluations; 2. A shorter development cycle, as marketed drugs have comprehensive preclinical pharmacokinetic and toxicology parameters, significantly shortening the development cycle for new indications based on these previous studies; 3. Lower development costs, significantly reducing R&D costs during preclinical and Phase I and II clinical trials compared to developing a new drug for the same indication.

[0037] In order to explore the strategy of "new uses of old drugs" for head and neck squamous cell carcinoma, the present invention selected 2248 small molecule compounds, mainly drugs approved by the U.S. Food and Drug Administration (1800 / 2248), and also included 319 clinical trial evaluation drugs and 129 preclinical compounds. Non-antitumor drugs accounted for more than 50% (1419 / 2248), and the remaining drugs were 718 targeted drugs and 111 therapeutic drugs. Inhibitors of common oncogene targets were included, including PI3K inhibitors, mTOR inhibitors, CDK inhibitors, and HDAC inhibitors, etc., covering a total of 286 different drug targets.

[0038] Subsequently, the present invention selected 13 primary tumor cells (PDCs) from patients and screened 2248 compounds with a single dose (1 μM) for three times, and measured the cell proliferation activity after 72 hours of drug action ( Figure 1Based on the initial screening results, 300 drugs that can cause more than 50% cell killing effect in more than one cell model were selected and screened at three concentrations (5μM, 1μM, 0.2μM) in five primary tumor cell lines ( Figure 2 ).

[0039] A total of 171 compounds were excluded because they had no concentration gradient dependence or could not reduce the number of viable cells by at least 50% in more than one cell model. Finally, 129 compounds were screened as drugs with good anti-head and neck squamous cell carcinoma activity, which formed the "old drug new use" collection of this invention.

[0040] Example 4: Pharmacogenomic research on the "old drug new use" collection to screen for drug efficacy sensitive biomarkers

[0041] This study used a 129-drug "old drug new use" collection to analyze the response curves of 54 head and neck squamous cell carcinoma cell lines (including 40 primary tumor cells and 14 commercial cell lines) over a 10-dose gradient. After 72 hours of drug treatment, the R package GRmetrics was used to calculate the drug response metrics IC50, Emax, and AUC, as well as the corresponding GR50, GRmax, and GRAOC.

[0042] For pharmacogenomic analysis, all primary tumor cells and commercially available cell lines underwent whole-exome and transcriptome sequencing. To identify signatures associated with drug response and develop predictive models, pharmacodynamic data were combined with genomic alterations, copy number variations, and gene expression signatures to assess their contribution to drug response. The potential of each marker as a biomarker to guide drug use was characterized. Predictive performance was evaluated using Spearman correlation and P-value.

[0043] Example 5 Pharmacogenomic analysis predicts that mitoxantrone is highly sensitive to head and neck squamous cell carcinoma with high IL6R expression

[0044] After analyzing the above 129 drugs for head and neck squamous cell carcinoma, we found that mitoxantrone (CAS: 70476-82-3, molecular formula: C 22 H 30 Cl2N4O6 (purchased from Selleck) has the highest efficacy among non-targeted drugs and has great potential for new uses in head and neck squamous cell carcinoma. In light of this, the present invention conducted a pharmacogenomic analysis of this marker. Table 1 shows potential candidate markers selected based on their biological function.

[0045] Table 1 Results of the potential of each gene as a biomarker to guide the use of mitoxantrone

[0046] Predictive markers Drug name p-value SNRPD3 Mitoxantrone <![CDATA[2.45E -10 ]]> AHSA1 Mitoxantrone <![CDATA[1.90E -09 ]]> IL16 Mitoxantrone <![CDATA[1.86E -08 ]]> KLF7 Mitoxantrone <![CDATA[1.72E -07 ]]> HECA Mitoxantrone <![CDATA[4.24E -06 ]]> SEMA4B Mitoxantrone <![CDATA[1.13E -05 ]]> SUMO4 Mitoxantrone <![CDATA[1.40E -05 ]]> RUNX2 Mitoxantrone <![CDATA[3.04E -05 ]]> PPT1 Mitoxantrone <![CDATA[3.04E -05 ]]> IL6R Mitoxantrone 0.000372348 FGF18 Mitoxantrone 0.000427904 GPR157 Mitoxantrone 0.000546867 ATF5 Mitoxantrone 0.000696705 AURKAIP1 Mitoxantrone <![CDATA[3.06E -08 ]]> RHOF Mitoxantrone 0.000713372

[0047] The present invention then selected 6 independent primary tumor cells and 4 commercial cell lines that had not been subjected to drug screening to form a validation cell set for mitoxantrone efficacy testing, and divided the cells into mitoxantrone-sensitive and resistant cells ( Figure 3 ).like Figure 3 As shown, HN13, PDC_54, PDC_55, and PDC_56 are mitoxantrone-resistant cells, and the rest are mitoxantrone-sensitive cells.

[0048] The present invention then tested the expression of 7 drug efficacy markers in the above 10 cell lines, and the results were as follows: Figure 4 As shown in the results, high expression of IL6R was significantly correlated with the sensitivity of mitoxantrone (P=0.049).

[0049] Example 6 In vitro experiments confirmed that mitoxantrone has a higher sensitivity to IL6R-highly expressed head and neck squamous cell carcinoma cells

[0050] Small interfering RNA design and synthesis:

[0051] Genetron Health Biotechnology Co., Ltd. was commissioned to synthesize the target small interfering RNA sequence. The specific small interfering RNA sequence is shown in Table 2.

[0052] Table 2 Small interfering RNA sequences

[0053] sense(5'-3') antisense(5'-3') si-IL6R-1 CCUCAGCAAUGUUGUUUGUTT ACAAACAACAUUGCUGAGGTT si-IL6R-2 GGCACUUACUACUAAUAAATT UUUAUUAGUAGUAAGUGCCTT si-IL6R-3 GCCCUUAUGACAUCAGCAATT UUGCUGAUGUCAUAAGGGCTT si-NC UUCUCCGAACGUGUCACGUTT ACGUGACACGUUCGGAGAATT

[0054] Small interfering RNA transfection:

[0055] The day before transfection, seed the target cells (primary tumor cells PDC_52, and commercial cell lines HSC3 and TU686) in each well of the well plate, and put 2.5mL of growth medium without antibiotics in each well. Remove the growth medium from the cells and add 1.5mL of fresh serum-free growth medium. Add 100pmol of small interfering RNA to 250μL of serum-free growth medium and mix gently. Add 5μL of transfection reagent to 250μL of growth medium for dilution and incubate at room temperature for 5min. Mix the small interfering RNA and transfection reagent diluted in the first two steps and incubate at room temperature for 20min. Add the mixture to the well plate containing cells seeded the day before. Culture the cells in a 37°C incubator for 5-6h. Replace the serum-containing medium and culture the cells for 24-48h before performing other detection steps after transfection. Detect protein expression after 3-4 days of culture.

[0056] Real-time PCR:

[0057] In order to detect the knockdown efficiency of the target gene, this example extracts total RNA from each group of cells, uses the mRNA as a template, and uses Oligo (dT) or random primers to reverse transcribe it into cDNA using reverse transcriptase. Then, PCR amplification is performed using the cDNA as a template to obtain the expression level of the IL6R gene. The results show that compared with the control group, the expression of the target gene in the small interfering RNA treatment group is significantly reduced ( Figure 5 ).

[0058] Cell proliferation assay:

[0059] Head and neck squamous cell carcinoma cells were seeded into 96-well plates at a density of 3000 cells / well and cultured in an incubator overnight. Subsequently, small interfering RNA was transfected and then treated with mitoxantrone. After 72 hours, cell proliferation was detected using the Cell Counting Kit-8. The results showed that compared with the IL6R knockdown group, mitoxantrone significantly inhibited the proliferation of the control group cells, indicating that IL6R-overexpressing head and neck squamous cell carcinoma cells are more sensitive to mitoxantrone ( Figure 6 ).

[0060] Example 7 In vivo transplant tumor experiments confirmed that mitoxantrone has a higher sensitivity to head and neck squamous cell carcinoma with high IL6R expression

[0061] Two head and neck squamous cell carcinoma PDX models (PDX_14 and PDX_31) were selected to evaluate the in vivo therapeutic effect of mitoxantrone on head and neck squamous cell carcinoma with different IL6R expression levels. Fifteen offspring models were constructed for each PDX model. When the tumor volume reached 100-200 mm 3 Then 10-12 mice were selected and divided into groups, namely, mitoxantrone drug treatment group and control group.

[0062] The dosing regimen is as follows:

[0063] ① Mitoxantrone treatment group: intravenous injection, 3 mg / kg, solvent is normal saline, twice a week.

[0064] ②Control group: intravenous injection of the corresponding drug solvent (normal saline), twice a week.

[0065] Continue to measure the model tumor volume and body weight until the tumor volume reaches 1000-1500mm 3 .

[0066] PDX_14 is a model with high expression of IL6R, with an expression level of FKPM=13.08. In the PDX_14 model, the mitoxantrone-treated group significantly inhibited tumor growth compared with the control group ( Figure 7PDX_31 is a model with low IL6R expression, with an expression level of FKPM = 6.48. In the PDX_31 model, the mitoxantrone drug treatment group had no significant effect on tumor growth compared to the control group ( Figure 8 ). It can be seen that in the xenograft tumor model, the IL6R expression level FKPM>13.08, which can be regarded as a high expression of IL6R in this model.

[0067] In some embodiments, the present invention further provides a kit for detecting the sensitivity of head and neck squamous cell carcinoma patients to mitoxantrone, wherein the kit comprises at least a reagent for detecting the expression level of IL6R, and the expression level of IL6R in head and neck squamous cell carcinoma tissue can be detected by RT-PCR or immunohistochemistry. Wherein, when the expression level of IL6R in head and neck squamous cell carcinoma tissue is detected by RT-PCR, the reagent comprises a nucleic acid that binds to the IL6R gene, and the nucleic acid comprises a primer for amplifying the IL6R gene. Wherein, when the expression level of IL6R in head and neck squamous cell carcinoma tissue is detected by immunohistochemistry, the reagent comprises a substance that binds to the IL6R protein, and the substance comprises an antibody that specifically binds to the IL6R protein.

[0068] In summary, the present invention has discovered for the first time that IL6R can serve as a biomarker for mitoxantrone in the treatment of head and neck squamous cell carcinoma (HNSCC). The higher the expression of IL6R, the more sensitive the HNSCC cells are to mitoxantrone. Based on this, the present invention provides a new use of mitoxantrone: mitoxantrone can be used to prepare a drug for treating HNSCC with high IL6R expression, and this has been experimentally verified both in vivo and in vitro. Furthermore, the present invention also provides a kit for detecting the sensitivity of HNSCC patients to mitoxantrone. This kit can be used to pre-test whether HNSCC patients are suitable for mitoxantrone treatment before treatment, thereby enabling the development of personalized treatment plans for different patients, improving treatment efficacy and quality of life.

[0069] Although the present invention has been described in detail through the above preferred embodiments, it should be understood that the above description is not intended to limit the present invention. After reading the above description, various modifications and substitutions of the present invention will become apparent to those skilled in the art. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. Use of a reagent for detecting IL6R expression level in the preparation of a kit for detecting the sensitivity of head and neck squamous cell carcinoma patients to mitoxantrone, characterized in that: The reagent for detecting the expression level of IL6R includes a nucleic acid that binds to the IL6R gene, or an antibody that specifically binds to the IL6R protein.

2. Use of the reagent for detecting IL6R expression level according to claim 1 in preparing a kit for detecting the sensitivity of head and neck squamous cell carcinoma patients to mitoxantrone, characterized in that: The nucleic acid includes primers for amplifying the IL6R gene.

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