TRPV4, application of TRPV4 inhibitor, head and neck cancer medicine and in-vitro non-therapeutic inhibition method

Abstract

The invention relates to application of TRPV4 and an inhibitor thereof, a head and neck cancer medicine and an in-vitro non-therapeutic inhibition method. According to the application of the TRPV4 as a head and neck cancer treatment target, the TRPV4 protein is remarkably and highly expressed in head and neck cancer tissues, and proliferation and migration of head and neck cancer cells can be inhibited and apoptosis can be promoted by knocking down the expression of the TRPV4 protein. The invention provides a novel therapeutic target for the head and neck cancer, and the target can be effectively used for judging and / or treating the head and neck cancer, so that a novel diagnostic and / or therapeutic agent is provided for the field and has a clinical application prospect.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and provides the use of TRPV4 and its inhibitor, a head and neck cancer drug, and an in vitro non-therapeutic inhibition method. BACKGROUND

[0002] Head and neck cancer is a general term for a type of cancer, including tumors in the oral cavity, larynx and nasopharynx, and is the seventh most common malignant tumor in the world. At present, the main treatment methods for head and neck cancer include surgery, radiotherapy, targeted therapy and immunotherapy. Although great progress has been made in the treatment of head and neck cancer, the 5-year overall survival rate of advanced head and neck cancer is still low, about 50%, which brings great challenges to clinical treatment. Therefore, it is crucial to explore new therapeutic targets for head and neck cancer.

[0003] In recent years, studies have found that the transient receptor potential (TRP) channel superfamily is a widely distributed non-selective cation channel located on the cell membrane, which is involved in the maintenance and regulation of intracellular Ca 2+ Homeostasis. According to its structural homology, it can be divided into seven subfamilies: TRPC (Canonical), TRPV (Vanilloid), TRPM (Melastatin), TRPML (Muclopins), TRPP (Polycystin), TRPA (Ankyrin) and TRPN (nompC). TRPV4 is a non-selective cation channel mainly through calcium ions, which belongs to the member of the vanilloid subfamily (TRPV). TRPV4 is widely expressed in various cells (such as nerve cells, endothelial cells and epithelial cells) and various tissues (such as lung, heart and kidney). The main functions of TRPV4 include the regulation of bone and cartilage development, the maintenance of body osmotic pressure, the perception of pain, and the regulation of vascular tension response to blood flow shear force. The ability of cell division and proliferation depends on Ca 2+ , which is required at the beginning of G1 phase, and during the transition from G1 / S and G1 / M. 2+ SUMMARY

[0004] Applicants found that abnormalities in calcium ion signals are closely related to the malignant progression of tumors. In head and neck cancer cells, abnormal activation of calcium ion signals can lead to abnormal expression of cell cycle-related proteins, thereby promoting the proliferation of tumor cells. TRPV4 (Transient Receptor Potential Vanilloid Receptor 4) is a non-specific cation channel with high permeability to calcium ions, which can mediate calcium ion influx.

[0005] The application is implemented by the following technical solutions:

[0006] ​The primary objective of this invention is to provide an application of TRPV4 as a therapeutic target for head and neck cancer. TRPV4 protein is significantly overexpressed in head and neck cancer tissues, and knocking down TRPV4 protein expression can inhibit the proliferation and migration of head and neck cancer cells and promote apoptosis.

[0007] The second objective of this invention is to provide the use of TRPV4 inhibitors in the preparation of medicaments for the prevention and / or treatment of head and neck cancer.

[0008] In one embodiment of the present invention, the inhibitor is a TRPV4 protein inhibitor or a TRPV4 gene inhibitor.

[0009] As one embodiment of the present invention, the TRPV4 protein inhibitor is selected from antibodies against the TRPV4 protein and / or binding proteins of the TRPV4 protein.

[0010] As one embodiment of the present invention, the TRPV4 gene inhibitor is selected from one or more of TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

[0011] The third objective of this invention is to provide a medicament for the prevention and / or treatment of head and neck cancer, comprising a pharmaceutically acceptable carrier and an effective amount of an active ingredient, said active ingredient being a TRPV4 protein inhibitor and / or a TRPV4 gene inhibitor.

[0012] As one embodiment of the present invention, the TRPV4 protein inhibitor is selected from antibodies against the TRPV4 protein and / or binding proteins of the TRPV4 protein.

[0013] As one embodiment of the present invention, the TRPV4 gene inhibitor is selected from one or more of TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

[0014] The fourth objective of this invention is to provide a method for in vitro non-therapeutic inhibition of head and neck cancer cells, wherein head and neck cancer cells are cultured in the presence of a TRPV4 protein inhibitor or a TRPV4 gene inhibitor.

[0015] In one embodiment of the present invention, the head and neck cancer cells are squamous cells of head and neck cancer.

[0016] The beneficial effects of this invention are: this invention provides a new therapeutic target for head and neck cancer, which can be effectively used for the diagnosis and / or treatment of head and neck cancer, thereby providing a novel diagnostic and / or therapeutic agent in the field with promising clinical application prospects. Attached Figure Description

[0017] Figure 1 TRPV4 was significantly highly expressed in head and neck cancer tissues. Figure 1 Image A is a representative image of TRPV4 staining in head and neck cancer (laryngeal cancer) tissue and adjacent tissue. Figure 1 B represents the statistical results of TRPV4 expression. LC: laryngeal cancer tissue, Ctrl: adjacent normal tissue. *P<0.05. Compared with the Ctrl group, the statistical method was Student's st-test.

[0018] Figure 2 The results were validated for the stable translocation of TRPV4-expressing HN4 cells; Figure 2 Image A is a representative image of TRPV4 expression in HN4 cells after overexpression. Figure 2 B represents the statistical results of TRPV4 expression in HN4 cells after overexpression. Figure 2 C is a representative image of TRPV4 expression in HN4 cells after knockdown. Figure 2 D represents the statistical results of TRPV4 expression in HN4 cells after knockdown. Figure 2 E is an image showing the calcium concentration in HN4 cells after overexpression / knockdown.

[0019] Figure 3 High expression of TRPV4 promotes the malignant progression of head and neck cancer cells; Figure 3 A represents the statistical results of HN4 cell proliferation after CCK-8 assay at different TRPV4 expression levels. Figure 3 B shows representative HN4 cell migration images after different TRPV4 expression levels were detected by cell scratch assay. Figure 3 Statistical results of representative HN4 cell migration after different TRPV4 expression levels. Figure 3 D shows representative images of HN4 cell apoptosis after TUNEL assay at different TRPV4 expression levels. Figure 3 Statistical results of HN4 cell apoptosis after different TRPV4 expression levels.

[0020] Figure 4 TRPV4 inhibitors can effectively block the malignant progression of head and neck cancer mediated by TRPV4 overexpression; Figure 4 A represents the statistical results of HN4 cell proliferation after the addition of a specific inhibitor following CCK-8 assay for TRPV4 overexpression. Figure 4 Image B shows a representative image of tumor size after adding a specific inhibitor following detection of TRPV4 overexpression in animal experiments. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise defined, the technical terms used in the following embodiments have the same meaning as commonly understood by those skilled in the art to which this invention pertains. Unless otherwise specified, the experimental reagents used in the following embodiments are all conventional biochemical reagents; the experimental methods, unless otherwise specified, are all conventional methods.

[0022] Unless otherwise specified, all experimental materials used in the following examples were purchased from conventional biochemical reagent stores. The present application is described in detail below with reference to specific embodiments.

[0023] Example 1

[0024] Studies have found that TRPV4 protein is significantly overexpressed in head and neck cancer tissues, and that high TRPV4 expression significantly enhances proliferation and growth. Knockdown of TRPV4 expression significantly inhibited the proliferation, growth, and migration of head and neck cancer cells and promoted apoptosis. Based on this, the invention of using TRPV4 as a therapeutic target for head and neck cancer was completed.

[0025] This invention provides the use of TRPV4 inhibitors in the preparation of medicaments for the prevention and / or treatment of head and neck cancer. The inhibitor is a TRPV4 protein inhibitor or a TRPV4 gene inhibitor, as the active ingredient.

[0026] Among them, the TRPV4 protein inhibitor is selected from antibodies against TRPV4 protein and / or binding proteins of TRPV4 protein; the TRPV4 gene inhibitor is selected from one or more of TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

[0027] Specifically, this application may use, but is not limited to, the following TRPV4 proteins or gene inhibitors:

[0028] TRPV4 protein inhibitors:

[0029] Antibodies against TRPV4 protein: anti-TRPV4 (brand: Abcam, catalog number: ab307444) (brand: CellSignaling Technology, catalog number: 65893); TRPV4 protein binding proteins: GSK2193874 (brand: MedChemExpress, catalog number: HY-100720), HC-067047 (brand: MedChemExpress, catalog number: HY-100208), RN1734 (brand: MedChemExpress, catalog number: HY-19975);

[0030] TRPV4 gene inhibitors:

[0031] TRPV4 siRNA and shRNA plasmids (brand: Santa Cruz Biotechnology, catalog number: sc-425650); TRPV4 gene-specific microRNA: miR-203; inhibitors that suppress the TRPV4 gene promoter: FOXC1 siRNA and shRNA plasmids (brand: Santa Cruz Biotechnology, catalog number: sc-43766).

[0032] This application provides a medicament for the prevention and / or treatment of head and neck cancer, comprising a pharmaceutically acceptable carrier and an effective amount of an active ingredient, said active ingredient being a TRPV4 protein inhibitor and / or a TRPV4 gene inhibitor.

[0033] Among them, the TRPV4 protein inhibitor is selected from antibodies against TRPV4 protein and / or binding proteins of TRPV4 protein; the TRPV4 gene inhibitor is selected from one or more of TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

[0034] The following TRPV4 proteins or gene inhibitors may be used, including but not limited to:

[0035] TRPV4 protein inhibitors:

[0036] Antibodies against TRPV4 protein: anti-TRPV4 (brand: Abcam, catalog number: ab307444) (brand: CellSignaling Technology, catalog number: 65893); TRPV4 protein binding proteins: GSK2193874 (brand: MedChemExpress, catalog number: HY-100720), HC-067047 (brand: MedChemExpress, catalog number: HY-100208), RN1734 (brand: MedChemExpress, catalog number: HY-19975);

[0037] TRPV4 gene inhibitors:

[0038] TRPV4 siRNA and shRNA plasmids (brand: Santa Cruz Biotechnology, catalog number: sc-425650); TRPV4 gene-specific microRNA: miR-203; inhibitors that suppress the TRPV4 gene promoter: FOXC1 siRNA and shRNA plasmids (brand: Santa Cruz Biotechnology, catalog number: sc-43766).

[0039] The term "effective amount" or "effective dose" refers to an amount that is functional or active in humans and / or animals and is acceptable to humans and / or animals.

[0040] The term "pharmaceuticalally acceptable" refers to a substance suitable for human and / or mammalian use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. The term "pharmaceuticalally acceptable carrier" refers to a carrier used for the administration of therapeutic agents, including various excipients and diluents.

[0041] Pharmaceutically acceptable carriers include, but are not limited to: water, saline, buffer solutions, glycerol, ethanol, liposomes, lipids, proteins, protein-antibody conjugates, peptides, cellulose, nanogels, or combinations thereof. The choice of carrier should be appropriate for the route of administration, as is well known to those skilled in the art.

[0042] The pharmaceutical compositions of the present invention contain a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier. Generally, pharmaceutical formulations should be matched to the route of administration; the dosage forms of the pharmaceutical compositions of the present invention are injections, oral formulations (tablets, capsules, oral liquids), transdermal formulations, and sustained-release formulations. They are prepared, for example, using physiological saline or an aqueous solution containing glucose and other excipients by conventional methods. The pharmaceutical compositions are preferably manufactured under aseptic conditions.

[0043] The effective amount of the active ingredient described in this invention can vary depending on the administration method and the severity of the disease to be treated. A preferred effective amount can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration.

[0044] A method for non-therapeutic inhibition of head and neck cancer cells in vitro is also provided, which involves culturing head and neck cancer cells in the presence of a TRPV4 protein inhibitor or a TRPV4 gene inhibitor. Head and neck cancer cells are squamous cells associated with head and neck cancer.

[0045] This application also provides specific experiments, taking the TRPV4 protein inhibitor HC-067047 as an example.

[0046] Example 2

[0047] I. Materials and Methods

[0048] 1. Experimental Materials

[0049] (1) Human head and neck cancer cells (HN4) were obtained from the Department of Otolaryngology, Atan Medical Center, Ulsan University College of Medicine, Seoul, South Korea (Kimetal., Acta Otolaryngol. 1997).

[0050] (2) Male BALB / c-nu mice (18-20g) were obtained from the Guangdong Provincial Medical Experimental Animal Center.

[0051] (3) TRPV4 overexpression lentiviral plasmid (purchased from Shanghai Heyuan Biotechnology Co., Ltd., name: pSLenti-SFH-EGFP-P2A-Puro-CMV-Trpv4-3xFLAG-WPRE), TRPV4 knockdown lentiviral plasmid (purchased from Shanghai Heyuan Biotechnology Co., Ltd., name: pSLenti-U6-shRNA(Trpv4)-CMV-EGFP-F2A-Puro-WPRE).

[0052] (4) Anti-TRPV4 antibody was purchased from Jiangsu Qinke Biotechnology Research Center Co., Ltd., catalog number: DF8624; anti-GAPDH antibody was purchased from Jiangsu Qinke Biotechnology Research Center Co., Ltd., catalog number: AF7021; Fluo-4AM was purchased from MedChemExpress, catalog number: HY-101896; TUNEL apoptosis detection kit was purchased from MedChemExpress, catalog number: HY-K1078; DAPI staining solution was purchased from Beyotime Biotechnology Co., Ltd., catalog number: C1006; CellCounting Kit-8 (CCK-8) was purchased from MedChemExpress, catalog number: HY-K0301; high-concentration matrix gel was purchased from MedChemExpress, catalog number: HY-K6008; TRPV4 inhibitor HC-067047 was purchased from MedChemExpress, catalog number: HY-100208.

[0053] 2. Experimental Methods

[0054] (1) HN4 cell proliferation was detected by CCK-8 assay. The specific steps are as follows: HN4 cells were injected with 2.0 × 10⁻⁶ cells per ... 4 / cm 2 Cells were seeded at a density in 96-well plates and cultured for 24 hours. The TRPV4 inhibitor group was added at a concentration of 50 nM and treated for 24 hours. Cell viability was assessed using the CellCount Kit-8 (CCK-8). Briefly, 10 μl of CCK-8 in 100 μl of culture medium was added to each well of cells, and the cells were incubated at 37°C for 1 hour. The amount of formazan dye produced was proportional to the number of viable cells and was detected at an absorbance wavelength of 450 nm, and quantified using an automated microplate reader (Rayto, Shandong, China).

[0055] (2) TRPV4 expression was detected by Western blotting. The specific steps are as follows:

[0056] 1) Add cells to RIPA lysis buffer (Shanghai Beyotime Biotechnology Co., Ltd.), which contains 1X PMSF and 1X protease and phosphatase inhibitor (Shanghai Beyotime Biotechnology Co., Ltd.). Incubate on ice for 30 min, centrifuge at 12000g for 20 min at 4℃, and collect the supernatant into a new 1.5ml centrifuge tube.

[0057] 2) Total protein concentration was determined by the BCA method (Shanghai Beyotime Biotechnology Co., Ltd.).

[0058] 3) 10% SDS-PAGE electrophoresis was used to detect the expression of TRPV4 and GAPDH, with a loading amount of 20 μg.

[0059] 4) Transfer membrane: Electrotransfer the proteins in the PAGE onto a PVDF membrane (Millipor, USA).

[0060] 5) Block with 5% skim milk at room temperature for 1 hour, then wash three times with PBST for 5 minutes each time.

[0061] 6) Primary antibody incubation: the anti-TRPV4 antibody was diluted 1:1000 and the anti-GAPDH antibody was diluted 1:2000. Incubate overnight at 4°C. Wash three times with PBST, 5 min each time.

[0062] 7) Secondary antibody incubation: goat anti-rabbit IgG-HRP (Jiangsu Qinke Biotechnology Research Center Co., Ltd.), incubated at room temperature for 2 hours, washed three times with PBST, 5 minutes each time.

[0063] 8) Exposure: Mix ECL developer (Tanon, Shanghai) 1:1 and add it evenly to the strip for exposure and color development.

[0064] (3) The expression level of TRPV4 in head and neck cancer (laryngeal cancer) tissues and adjacent normal tissues was detected by immunohistochemistry. The specific steps are as follows:

[0065] 1) Head and neck cancer patient tissues and adjacent tissues were placed in 4% paraformaldehyde and fixed at room temperature for 48 hours before dehydration, embedding, and sectioning.

[0066] 2) Dewaxing: Xylene 15min → Xylene 15min → Anhydrous ethanol 5min → Anhydrous ethanol 5min → 90% ethanol 5min → 80% ethanol 5min → 70% ethanol 5min → Pure water 5min.

[0067] 3) Thermal repair (pH 6.0 citric acid repair solution): Microwave repair for 30 minutes, then cool to room temperature.

[0068] 4) Cover the tissue with goat serum (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.) and incubate at 37℃ for 30 min.

[0069] 5) Primary antibody incubation: The anti-IGF1R antibody is diluted 1:200. Cover and incubate overnight at 4°C. Wash three times with PBS, 5 min each time.

[0070] 6) Secondary antibody incubation: Cover the tissue with the working solution of the secondary antibody, incubate at 37°C for 30 min, and wash three times with PBS for 5 min each time.

[0071] 7) Dilute DAB to 1X, prepare fresh and use immediately, cover the tissue, and develop color.

[0072] 8) Hematoxylin counterstaining, gradient dehydration, and clearing and mounting.

[0073] (4) Cell migration was detected by cell scratch assay. The specific steps are as follows:

[0074] 1) When HN4 cells with different TRPV4 expression levels reach 100% confluence, use a 10μl pipette tip to draw a cross shape.

[0075] 2) Wash away the cell debris with PBS and replace with serum-free culture medium, and record the time as 0h.

[0076] 3) Take another photo at the same location 36 hours later to compare the degree of cell healing after 36 hours.

[0077] (5) Cell apoptosis was detected by TUNEL assay. The specific steps are as follows:

[0078] 1) Slide climbing: HN4 cells with different TRPV4 expression levels were climbed and cultured for 24 h.

[0079] 2) Fixation: Wash three times with PBS, 5 minutes each time, and then place in 4% paraformaldehyde for 30 minutes at room temperature.

[0080] 3) Permeabilization: Wash 3 times with PBS for 5 minutes each time, cover and incubate with 0.1% Triton X-100 (dissolved in PBS) for 5 minutes, then digest and inactivate endogenous peroxidase with 2 mg / ml proteinase K (incubate at 37°C for 15 minutes).

[0081] 4) Equilibration: Add equilibration solution and incubate at 37°C for 30 minutes.

[0082] 5) Staining: The reaction solution of terminal deoxynucleotidyl transferase (TdT) and FITC-dUTP was incubated at 37°C in the dark for 60 minutes.

[0083] 6) Counterstaining: Cover with DPAI staining solution and let stand at room temperature for 5 minutes.

[0084] 7) Mounting and Photography: Mount the slides with anti-fluorescence quenching mounting medium (Shanghai Beyotime Biotechnology Co., Ltd.), and after drying, analyze using a fluorescence microscope. Blue fluorescence at 460 nm (DAPI) reveals each cell, while green fluorescence at 520 nm identifies apoptotic cells. Apoptosis percentage = (Number of apoptotic cells / Total number of cells) × 100%.

[0085] (6) The growth of head and neck cancer was detected using a xenograft mouse model (CDX). The specific steps are as follows:

[0086] HN4 cells (with different TRPV4 expression levels) (1×10⁶ cells) suspended in 50 μL DMEM high-glucose medium were mixed with Matrigel (1:1) (MedChemExpress, USA) and subcutaneously injected into mice. Tumor size was observed in mice after 2 weeks. The TRPV4 inhibitor group received intraperitoneal injection of HC-067047 at a concentration of 50 mg / kg, once every 3 days for 1 month.

[0087] 3. Statistical Methods

[0088] The software GraphPadPrism5 was used; the Student's st-test was used to compare the means of two groups.

[0089] II. Experimental Results

[0090] 1. TRPV4 is significantly overexpressed in head and neck cancer patient tissues.

[0091] The results are as follows Figure 1 As shown in A-1B, representative images and statistical results of TRPV4 expression in head and neck cancer (laryngeal cancer) tissues are presented. The results show that TRPV4 PCNA expression is significantly increased in head and neck cancer tissues compared to adjacent normal tissues. These results suggest that increased TRPV4 expression may be an important cause of malignant progression in head and neck cancer.

[0092] 2. Screening and validation of HN4 cells with TRPV4 overexpression or knockdown

[0093] The results are as follows Figure 2 As shown in A-2B, compared with the control group, the TRPV4 expression level in HN4 cells in the TRPV4 OE group was significantly increased; Figure 2 As shown in C-2D, compared with the control group, the expression level of TRPV4 in HN4 cells in the TRPV4 KD group was significantly reduced; Figure 2 As shown in Figure E, compared with the control group, the intracellular calcium ion concentration of HN4 cells in the TRPV4 OE group was increased, while the intracellular calcium ion concentration of HN4 cells in the TRPV4 KD group was decreased. These results suggest that HN4 cells of TRPV4 OE and TRPV4 KD have been successfully screened.

[0094] 3. High expression of TRPV4 significantly promotes the malignant progression of head and neck cancer cells.

[0095] like Figure 3 As shown in Figure A, TRPV4OE significantly promoted the proliferation of HN4 cells, while TRPV4KD significantly inhibited the proliferation of HN4 cells. Figure 3 B-3C showed that TRPV4 OE significantly promoted the migration of HN4 cells, while TRPV4 KD significantly inhibited the migration of HN4 cells. Figure 3The D-3E results also showed that TRPV4 OE significantly inhibited HN4 cell apoptosis, while TRPV4KD significantly promoted HN4 cell apoptosis. Figure 3 The results showed that, compared with the CDX model established by HN4, the CDX model established by TRPV4 OE-HN4 exhibited significantly increased tumor growth, while the CDX model established by TRPV4 KD-HN4 showed significantly decreased tumor growth. These results suggest that high expression of TRPV4 is an important factor promoting the growth of head and neck cancer tumors.

[0096] 4. TRPV4 inhibitors can effectively block the malignant progression of head and neck cancer mediated by TRPV4 overexpression.

[0097] like Figure 4 As shown in Figure A, the TRPV4 OE-induced proliferation of HN4 cells can be blocked by the TRPV4-specific inhibitor (HC-067047). Figure 4 The results showed that the growth-promoting effect of TRPV4 OE on tumor tissue was significantly inhibited by HC-067047. These results suggest that TRPV4 inhibitors can significantly block the malignant progression of head and neck cancer induced by TRPV4 OE.

[0098] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of TRPV4 as a therapeutic target for head and neck cancer is characterized by: TRPV4 protein is significantly overexpressed in head and neck cancer tissues. Knocking down TRPV4 protein expression can inhibit the proliferation and migration of head and neck cancer cells and promote apoptosis.

2. Use of TRPV4 inhibitors in the preparation of drugs for the prevention and / or treatment of head and neck cancer.

3. The use according to claim 2, characterized in that: The inhibitor is a TRPV4 protein inhibitor or a TRPV4 gene inhibitor.

4. The use according to claim 3, characterized in that: The TRPV4 protein inhibitor is selected from antibodies against the TRPV4 protein and / or binding proteins of the TRPV4 protein.

5. The use according to claim 3, characterized in that: The TRPV4 gene inhibitor is selected from one or more of the following: TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

6. A drug for the prevention and / or treatment of head and neck cancer, characterized in that: It contains a pharmaceutically acceptable carrier and an effective amount of an active ingredient, said active ingredient being a TRPV4 protein inhibitor and / or a TRPV4 gene inhibitor.

7. The medicament for preventing and / or treating head and neck cancer according to claim 6, characterized in that: The TRPV4 protein inhibitor is selected from antibodies against the TRPV4 protein and / or binding proteins of the TRPV4 protein; the TRPV4 gene inhibitor is selected from one or more of the following: TRPV4 gene-specific RNAi, TRPV4 gene-specific microRNA, and inhibitors that inhibit the TRPV4 gene promoter.

8. The medicament for preventing and / or treating head and neck cancer according to claim 6, characterized in that: The drug dosage forms are injections, oral preparations, transdermal preparations, and sustained-release preparations.

9. A method for in vitro non-therapeutic inhibition of head and neck cancer cells, characterized in that: Head and neck cancer cells were cultured in the presence of TRPV4 protein inhibitors or TRPV4 gene inhibitors.

10. The method for in vitro non-therapeutic inhibition of head and neck cancer cells according to claim 9, characterized in that: The head and neck cancer cells are squamous cells of the head and neck cancer.

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