Application of biomarker in preparation of products for diagnosing and / or preventing and treating esophageal cancer diseases

Abstract

The invention discloses application of a biomarker in preparation of a product for diagnosing and / or preventing and treating esophageal cancer diseases. A verification result provided by the invention proves that interference of the CD24 gene can inhibit invasion and metastasis of esophageal squamous carcinoma cells, and the CD24 gene has very important significance on treatment of esophageal squamous carcinoma. The verification result provided by the invention also proves that the interference of the CD24 gene obviously enhances the sensitivity of the esophageal squamous carcinoma cells to the 5-fluorouracil, and obviously reduces the clone formation ability of the esophageal squamous carcinoma cells during 5-FU treatment, and the CD24 inhibitor and the 5-fluorouracil can be used in combination for clinical treatment of the esophageal squamous carcinoma.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the application of biomarkers in the preparation of products for diagnosing and / or preventing and treating esophageal cancer. Background Art

[0002] Esophageal cancer (ESCA) is one of the most aggressive cancers and the sixth leading cause of cancer death worldwide, posing a serious threat and challenge to human health. Esophageal cancer mainly includes two pathological subtypes: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC), of which ESCC accounts for more than 90% of the annual incidence of esophageal cancer, and there are obvious differences in their geographical distribution and epidemiology.

[0003] The main methods for treating esophageal cancer in clinic include surgical resection, radiotherapy and chemotherapy. However, due to the limited efficacy and serious adverse reactions of conventional treatment, the results are still unsatisfactory. The main reasons are: 1. Surgical resection of esophageal cancer is only applied to patients with esophageal cancer in the early stage, while most patients are already in the middle and late stages when diagnosed, and the tumor has spread or metastasized, and cannot receive surgical treatment; 2. Chemotherapy, as another treatment method, is becoming increasingly important in the comprehensive treatment of tumors. However, cancer cells often develop resistance to chemotherapy drugs, which cannot kill cancer cells. Residual cancer cells will form new lesions, thereby causing tumor recurrence and metastasis. This is also the main reason why chemotherapy fails in many cases. Therefore, clarifying the molecular mechanism of the occurrence and development of esophageal cancer and identifying and confirming the key molecules in this process will provide valuable clues for the study of the mechanism of tumorigenesis, and will also provide new targets for the early diagnosis and treatment of esophageal cancer tumors, making an important contribution to further improving the level of esophageal cancer treatment.

[0004] CD24, or Cluster of differentiation 24, is a highly glycosylated glycosylphosphatidylinositol (GPI)-anchored membrane protein, and its relationship with the occurrence and development of esophageal cancer has not yet been clearly clarified. Summary of the invention

[0005] The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide an application of CD24 in the preparation of products for diagnosing and / or preventing and treating esophageal cancer.

[0006] The purpose of the present invention is achieved through the following technical solutions: Application of CD24 in the preparation of products for diagnosing and / or preventing and treating esophageal cancer.

[0007] Furthermore, the application is the use of CD24 inhibitors in the preparation of products for treating and / or preventing esophageal cancer.

[0008] Furthermore, the product for treating and / or preventing esophageal cancer is a drug that can inhibit the invasion and / or metastasis of esophageal cancer cells.

[0009] Furthermore, the metastasis includes but is not limited to lung metastasis.

[0010] Application of CD24 in the preparation of drugs for enhancing the sensitivity of esophageal cancer to chemotherapy.

[0011] Furthermore, the application is the use of the CD24 inhibitor in the preparation of a drug for enhancing the sensitivity of esophageal cancer to chemotherapy.

[0012] Furthermore, the chemotherapy drug includes 5-fluorouracil.

[0013] Application of CD24 inhibitor combined with 5-fluorouracil in the preparation of anti-esophageal cancer drugs.

[0014] Furthermore, the CD24 inhibitors include reagents used in gene editing technologies such as RNA interference technology, antisense oligonucleotide technology, CRISPR technology, TALEN technology, ZFN technology, Cre-loxP gene recombination technology, and also include compounds that can reduce the expression of CD24.

[0015] Furthermore, the CD24 inhibitor includes shRNA used in RNA interference technology, and its target sequence is: shCD24#1: 5'-CTTCTGCATCTCTACTCTTAA-3'; Or, shCD24#2: 5'-TGCTCCTACCCACGCAGATTT-3'.

[0016] Furthermore, the esophageal cancer includes but is not limited to esophageal squamous cell carcinoma.

[0017] Furthermore, the medicine also contains pharmaceutically acceptable excipients.

[0018] Furthermore, the pharmaceutically acceptable excipient is preferably at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant and a lubricant.

[0019] Furthermore, the administration methods of the drug for treating and / or preventing esophageal cancer include but are not limited to oral administration and injection administration.

[0020] Compared with the prior art, the present invention has the following advantages and effects: The validation results provided by the present invention indicate that CD24 has good diagnostic efficacy, and whether a subject has the risk of esophageal cancer metastasis or drug resistance can be diagnosed based on CD24, and has good clinical application value.

[0021] The verification results provided by the present invention prove that the interference of CD24 gene can inhibit the invasion and metastasis of esophageal squamous cell carcinoma cells, which is of great significance to esophageal squamous cell carcinoma and the treatment of esophageal squamous cell carcinoma.

[0022] The verification results provided by the present invention also prove that the interference of CD24 gene significantly enhances the sensitivity of esophageal squamous cell carcinoma cells to 5-fluorouracil, and significantly reduces the clone formation ability of esophageal squamous cell carcinoma cells when treated with 5-FU. CD24 inhibitors can be used in combination with 5-fluorouracil for the clinical treatment of esophageal squamous cell carcinoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a graph showing the effect of CD24 gene interference on the invasion / migration ability of human esophageal squamous cell carcinoma cells KYSE150-LUC-LM3 and human esophageal squamous cell carcinoma cells EC109.

[0024] Figure 2 This figure shows the effect of CD24 gene interference on the lung metastasis ability of human esophageal squamous cell carcinoma cell line KYSE150-LUC-LM3.

[0025] Figure 3 This is a graph showing the effects of different concentrations of 5-FU on the activity of esophageal squamous cell carcinoma cells that overexpress CD24.

[0026] Figure 4 This is a graph showing the effect of different concentrations of 5-FU on the clone-forming ability of esophageal squamous cell carcinoma cells that overexpress CD24.

[0027] Figure 5 This is a graph showing the effects of different concentrations of 5-FU on the activity of CD24 knockdown esophageal squamous cell carcinoma cells.

[0028] Figure 6 This is a graph showing the effect of different concentrations of 5-FU on the clone-forming ability of esophageal squamous cell carcinoma cells with CD24 knockdown. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0030] The human esophageal squamous cell carcinoma cells KYSE150, KYSE30, KYSE510, and EC109 used in the following examples were purchased from German Microbiology and Cell Culture Co., Ltd.; LB medium was purchased from Shanghai Bioengineering Co., Ltd., and 0.25% trypsin was purchased from Gibco, USA. Experimental female mice (NOD-SCID) were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0031] The human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC used in the following examples are described in “WenWen Xu et al. Direct Targeting of CREB1 with Imperatorin Inhibits TGFβ2-ERKSignaling to Suppress Esophageal Cancer Metastasis, Advanced Science, 2020. DOI: 10.1002 / advs.202000925”.

[0032] The human esophageal squamous cell carcinoma 5-FU-resistant (FR) cell lines KYSE150FR and KYSE410FR have been described in “BinLi et al. Competitive Binding Between Id1 and E2F1 to Cdc20 Regulates E2F1 Degradationand Thymidylate Synthase Expression to Promote Esophageal Cancer Chemoresistance, Clin Cancer Res. 2016. DOI: 10.1158 / 1078-0432.CCR-15-1196”.

[0033] The KYSE150-LUC-LM3 cell construction method used in the following examples is as follows: 0.25% trypsin was used to digest the human esophageal squamous cell carcinoma luciferase-labeled KYSE150-LUC cells into single cells. 6The LUCI-labeled KYSE150-LUC cells of human esophageal squamous cell carcinoma were resuspended in 100 µL PBS buffer and injected into the mice through the tail vein. After 5 to 6 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally for in vivo fluorescence photography. The mice with lung metastasis were dissected and the lungs with KYSE150-LUC cells were removed. The lungs were minced into minced meat and resuspended in culture medium. After standing for one week, the cells were washed twice with PSB buffer and Blasticidin (5 µg / mL, purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.) was added. The cells were screened for one week until there was no cell death. These cells were KYSE150-LUC-LM1. Then, 0.25% trypsin was used to digest the LUCI-labeled KYSE150-LUC-LM1 cells into single cells. 10 6 Individual esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC-LM1 were resuspended in 100 µL PBS buffer and injected into mice through the tail vein. After 5 to 6 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally and in vivo fluorescence photography was performed. Mice with lung metastasis were dissected and the lungs with KYSE150-LUC-LM1 cells were removed. The lungs were minced into minced meat and resuspended in culture medium. After standing for one week, the cells were washed twice with PBS buffer and Blasticidin (5 µg / mL, purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.) was added. The cells were screened for one week until there was no cell death. These cells were KYSE150-LUC-LM2. This cycle was repeated three times to obtain KYSE150-LUC-LM3 cells.

[0034] The construction methods of the human esophageal squamous cell carcinoma cell KYSE150-LUC-LM3 with stable knockdown of CD24 gene (i.e., KYSE150-LUC-LM3-shCD24 cells) and the human esophageal squamous cell carcinoma cell EC109 with stable knockdown of CD24 gene (i.e., EC109-shCD24 cells) used in the following examples are as follows: The CD24 interference fragment (shCD24 target sequence: shCD24#1: 5'-CTTCTGCATCTCTACTCTTAA-3'; shCD24#2: 5'-TGCTCCTACCCACGCAGATTT-3') was constructed on the pLKO.1-PuroR expression vector (commissioned by Shanghai Quanyang Biotechnology Co., Ltd.). The constructed plasmid was transformed into 293T cells, and the 293T cells with good growth status were digested and plated on a 6-well plate. When the cells grew to 50%-60%, CD24 knockdown plasmid transfection was performed. After 24 hours and 48 hours, the cells were replaced with medium and the supernatant was collected. The supernatant was centrifuged and filtered with a 0.45 µM filter membrane. The supernatant was added to KYSE150-LUC-LM3 cells and human esophageal cancer EC109 cells, respectively. After 48 hours of infection, the cells were transferred from the six-well plate to a vial. After adherence, puromycin (purchased from Sigma-Aldrich) at a working concentration of 1 µg / mL was added for screening. After 1-2 weeks, when the cells no longer died, the addition of drugs was stopped to obtain human esophageal squamous cell carcinoma cells KYSE150-LUC-LM3 with stable knockdown of CD24 gene (i.e., KYSE150-LUC-LM3-shCD24 cells) and human esophageal squamous cell carcinoma cells EC109 with stable knockdown of CD24 gene (i.e., EC109-shCD24 cells). Western blot was used to identify that the cells were able to interfere with CD24 expression.

[0035] The construction methods of the human esophageal squamous cell carcinoma cell KYSE30 (i.e., KYSE30-CD24 cell) with stable overexpression of the CD24 gene and the human esophageal squamous cell carcinoma cell KYSE510 (i.e., KYSE510-CD24 cell) with stable overexpression of the MEST gene used in the following examples are as follows: The CD24 gene (NCBI accession number: NM_001291737.1) was constructed on the pTSB-CMV-MCS-SBP-3Flag-copGFP-F2A-PuroR expression vector to obtain the CD24 eukaryotic expression plasmid (commissioned to Shanghai Quanyang Biotechnology Co., Ltd.). The constructed plasmid was transformed into 293T cells, and the 293T cells with good growth status were digested and plated on a 6-well plate. When the cells grew to 50%-60%, the CD24 eukaryotic expression plasmid was transfected. After 24 hours and 48 hours, the cells were replaced with liquid and the supernatant was collected. The supernatant was centrifuged and filtered with a 0.45 µM filter membrane. The supernatant was added to KYSE30 cells and human esophageal cancer KYSE510 cells, respectively. After 48 hours of infection, the cells were transferred from the six-well plate to a vial. After adherence, puromycin (purchased from Sigma-Aldrich) at a working concentration of 1 µg / mL was added for screening. After 1-2 weeks, when the cells no longer died, the drug addition was stopped to obtain human esophageal squamous cell carcinoma cells KYSE30 with stable overexpression of the CD24 gene (i.e., KYSE30-CD24 cells) and human esophageal squamous cell carcinoma cells KYSE510 with stable overexpression of the CD24 gene (i.e., KYSE510-CD24 cells). Western blot was used to identify that the cells were able to overexpress CD24.

[0036] Example 1: CD24 interference inhibits the metastasis and invasion ability of human esophageal squamous cell carcinoma cells 1. Knock down CD24 in esophageal squamous cell carcinoma cells, and then detect the cell invasion ability through invasion experiment, migration experiment and in vivo mouse experiment.

[0037] (1) Invasion experiment 1) Chamber treatment: Add 100 μL of 5% Matrigel to the clean invasion chamber and place it in a 37°C incubator for 30 min.

[0038] 2) Cell plating: Human esophageal squamous cell carcinoma cells KYSE150-LUC-LM3 with stable knockdown of CD24 gene (i.e., KYSE150-LUC-LM3-shCD24 cells) and human esophageal squamous cell carcinoma cells EC109 with stable knockdown of CD24 gene (i.e., EC109-shCD24 cells) were digested into suspended single cells with 0.25% trypsin, and after counting, 500,000 KYSE150-LUC-LM3-shCD24 cells and 200,000 EC109-shCD24 cells were added to the upper layer of each chamber, and 500 μL complete medium was added to the lower layer, and cultured at 37°C and 5% CO2 for 24 h. At the same time, human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC-LM3-shCON and human esophageal squamous cell carcinoma cells EC109-shCON were used as controls for the above experiment.

[0039] 3) Crystal violet staining: Take out the chamber, wash twice with PBS buffer (pH 7.4, purchased from Sigma-Aldrich; the same below), add 500 μL of methanol to the lower chamber, fix for 10 min, remove the methanol, add 0.2% crystal violet to stain for 10 min, remove the crystal violet and wash away the residual crystal violet with clean water, dry and take pictures.

[0040] (2) Migration experiment In the migration experiment, the invasion chamber was not treated with 5% Matrigel, and the other steps were the same as those of invasion.

[0041] (3) In vivo experiments Twelve 6-week-old female mice (NOD-SCID), 6 in the control group and 6 in the experimental group, were selected to construct a tumor metastasis model.

[0042] 1) Digest KYSE150-LUC-LM3-shCON and KYSE150-LUC-LM3-shCD24 cells, human esophageal squamous cell carcinoma luciferase-labeled cells, into single cells using 0.25% trypsin. Take 10 6 Resuspend cells in 100 µL PBS buffer.

[0043] 2) Anesthetize the mice before the tail vein experiment, and evaluate the degree of anesthesia through painless and painful stimulation to ensure that the mice are in an anesthetized state; 3) Resuspend the cells using a microsyringe with a 25G needle and inject into the tail vein of 12 mice.

[0044] After 5 to 6 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally, and in vivo fluorescence photography was performed to observe the lung metastasis of cancer cells.

[0045] The results are as follows Figure 1 and Figure 2 As shown. Figure 1 The results of in vitro experiments showed that CD24 interference could inhibit the invasion of human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC-LM3 and human esophageal squamous cell carcinoma cells EC109. Figure 2 The results of in vivo experiments showed that CD24 interference could inhibit the lung metastasis of human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC-LM3 in mice.

[0046] Example 2: CD24 promotes 5-FU chemotherapy resistance in human esophageal squamous cell carcinoma cells CD24 was stably overexpressed in esophageal squamous cell carcinoma cells, and then the biological function of CD24 during 5-FU chemotherapy of esophageal cancer was detected by CCK-8 experiment and clone formation experiment. The specific steps of CCK-8 experiment are as follows: 1. When the cells grow to 90%, discard the original culture medium of the cells, wash the cells twice with PBS buffer, add 0.25% trypsin, and place them in the incubator for digestion for a period of time.

[0047] 2. After digestion is complete, add an appropriate amount of complete medium to terminate digestion.

[0048] 3. Use a pipette to gently blow the surface of the cell culture dish and the cell suspension to disperse the cells, and transfer them to a clean centrifuge tube. Centrifuge at 1000 rpm for 3 min.

[0049] 4. Discard the supernatant, add an appropriate amount of complete culture medium to resuspend the cells, and count them.

[0050] 5. Inoculate the cells in a 96-well plate at a density of 3,000 cells per well (control group and experimental group). After addition, tap the edge of the 96-well plate three times in a clockwise direction to mix the cells. Let stand for about 5 minutes and then culture in a cell culture incubator overnight.

[0051] 6. After adhesion, different concentrations (0 μM, 5 μM, 10 μM, and 20 μM) of the drug 5-FU were added to treat CD24-overexpressing human esophageal squamous cell carcinoma cells KYSE30 and KYSE510 (i.e., KYSE30-CD24, KYSE510-CD24) and corresponding control cells (i.e., KYSE30-CON, KYSE510-CON) for 48 h.

[0052] 7. Aspirate the culture medium in the 96-well plate and add 100 μL of the prepared CCK-8 solution (CCK-8 solution: culture medium = 1:9) to each well, avoiding the formation of bubbles, and incubate in a cell culture incubator away from light for 1-4 h (depending on experimental requirements).

[0053] 8. Use an ELISA reader to measure the absorbance at 450 nm and calculate the inhibitory effect of 5-FU on each cell.

[0054] The specific steps of the clone formation experiment are as follows: 1. When the cells have grown to 90%, trypsinize, resuspend and count.

[0055] 2. After counting, 2000 cells were inoculated into 6-well plates. After adherence, different concentrations (0 μM, 0.625 μM, 1.25 μM, and 2.5 μM) of the drug 5-FU were added to treat CD24-overexpressing human esophageal squamous cell carcinoma cells KYSE30 and KYSE510 (i.e., KYSE30-CD24, KYSE510-CD24), as well as corresponding control cells (i.e., KYSE30-CON, KYSE510-CON) for 14 days, during which the medium was changed as needed.

[0056] 3. Discard the culture medium and add PBS buffer for washing.

[0057] 4. Add 1 mL of methanol and fix for 10 min.

[0058] 5. Discard the methanol and add crystal violet staining solution for 5 min.

[0059] 6. After rinsing with clean water, turn the 6-well plate upside down and scan and take pictures after drying.

[0060] The results are as follows Figure 3 and Figure 4 As shown, from Figure 3 The experimental results show that overexpression of CD24 significantly inhibited the sensitivity of human esophageal squamous cell carcinoma cells KYSE30 and KYSE510 to 5-FU. Figure 4 The experimental results show that overexpression of CD24 significantly improved the cloning ability of human esophageal squamous cell carcinoma cells KYSE30 and KYSE510 when treated with 5-FU.

[0061] Example 3: CD24 interference enhances 5-FU sensitivity in human esophageal squamous cell carcinoma cells CD24 was interfered with in esophageal squamous cell carcinoma cells, and then the biological function of CD24 in 5-FU chemotherapy of esophageal cancer was detected by CCK-8 experiment and clone formation experiment. The specific steps of CCK-8 experiment are as follows: 1. When the cells grow to 90%, discard the original culture medium of the cells, wash the cells twice with PBS buffer, add 0.25% trypsin, and place them in the incubator for digestion for a period of time.

[0062] 2. After digestion is complete, add an appropriate amount of complete medium to terminate digestion.

[0063] 3. Use a pipette to gently blow the surface of the cell culture dish and the cell suspension to disperse the cells, and transfer them to a clean centrifuge tube. Centrifuge at 1000 rpm for 3 min.

[0064] 4. Discard the supernatant, add an appropriate amount of complete culture medium to resuspend the cells, and count them.

[0065] 5. Inoculate the cells in a 96-well plate at a density of 3,000 cells per well (control group and experimental group). After addition, tap the edge of the 96-well plate three times in a clockwise direction to mix the cells. Let stand for about 5 minutes and then culture in a cell culture incubator overnight.

[0066] 6. After adhesion, different concentrations (0 μM, 5 μM, 10 μM, and 20 μM) of the drug 5-FU were added to treat CD24-knockdown human esophageal squamous cell carcinoma cells KYSE150FR and KYSE410FR (i.e., KYSE150FR-shCD24 and KYSE410FR-shCD24), as well as the corresponding control cells (i.e., KYSE150FR-shCON and KYSE410FR-shCON) for 48 hours.

[0067] 7. Aspirate the culture medium in the 96-well plate and add 100 μL of the prepared CCK-8 solution (CCK-8 solution: culture medium = 1:9) to each well, avoiding the formation of bubbles, and incubate in a cell culture incubator away from light for 1-4 h (depending on experimental requirements).

[0068] 8. Use an ELISA reader to measure the absorbance at 450 nm and calculate the inhibitory effect of 5-FU on each cell.

[0069] The specific steps of the clone formation experiment are as follows 1. When the cells have grown to 90%, trypsinize, resuspend and count.

[0070] 2. After counting, 2000 cells were inoculated into 6-well plates. After adherence, different concentrations (0 μM, 0.625 μM, 1.25 μM, and 2.5 μM) of the drug 5-FU were added to treat CD24-knockdown human esophageal squamous cell carcinoma cells KYSE150FR and KYSE410FR (i.e., KYSE150FR-shCD24 and KYSE410FR-shCD24), as well as corresponding control cells (i.e., KYSE150FR-shCON and KYSE410FR-shCON) for 14 days, during which the medium was changed as needed.

[0071] 3. Discard the culture medium and add PBS buffer for washing.

[0072] 4. Add 1 mL of methanol and fix for 10 min.

[0073] 5. Discard the methanol and add crystal violet staining solution for 5 min.

[0074] 6. After rinsing with clean water, turn the 6-well plate upside down and scan and take pictures after drying.

[0075] The results are as follows Figure 5 and Figure 6 As shown. Figure 5 The experimental results show that the interference of CD24 significantly enhances the sensitivity of human esophageal squamous cell carcinoma cells KYSE150FR and KYSE410FR to 5-FU. Figure 6 The experimental results show that CD24 interference significantly reduced the cloning ability of human esophageal squamous cell carcinoma cells KYSE150FR and KYSE410FR when treated with 5-FU.

[0076] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of CD24 in the preparation of products for diagnosing and / or preventing and treating esophageal cancer.

2. The use according to claim 1, characterized in that: The application is the use of CD24 inhibitors in the preparation of products for treating and / or preventing esophageal cancer.

3. The use according to claim 2, characterized in that: The product for treating and / or preventing esophageal cancer is a drug that can inhibit the invasion and / or metastasis of esophageal cancer cells; The metastasis includes lung metastasis.

4. Application of CD24 inhibitors in the preparation of drugs for enhancing the sensitivity of esophageal cancer to chemotherapy.

5. The use according to claim 4, characterized in that: The chemotherapy drugs include 5-fluorouracil.

6. Application of CD24 inhibitors combined with 5-fluorouracil in the preparation of anti-esophageal cancer drugs.

7. The use according to any one of claims 2 to 6, characterized in that: The CD24 inhibitor includes any one or more of reagents used in RNA interference technology, antisense oligonucleotide technology, CRISPR technology, TALEN technology, ZFN technology, Cre-loxP gene recombination technology and compounds that reduce the expression of CD24.

8. The use according to any one of claims 2 to 6, characterized in that: The CD24 inhibitor includes shRNA used in RNA interference technology, and its target sequence is: shCD24#1: 5'-CTTCTGCATCTCTACTCTTAA-3'; Or, shCD24#2: 5'-TGCTCCTACCCACGCAGATTT-3'.

9. The use according to any one of claims 2 to 6, characterized in that: The esophageal cancer includes esophageal squamous cell carcinoma.

10. The use according to any one of claims 2 to 6, characterized in that: The drug further comprises a pharmaceutically acceptable excipient; The pharmaceutically acceptable excipient is at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant and a lubricant; The administration method of the medicine for treating and / or preventing esophageal cancer is oral administration or injection administration.

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