Construction method and application of mouse gastric cancer spontaneous tumor model

Abstract

The invention provides a construction method and application of a mouse gastric cancer spontaneous tumor model, and the construction method comprises the following steps: reducing or inhibiting the expression quantity of a TP53 gene and / or the activity of a P53 protein in a mouse body, and then giving a drug to the mouse for chemical induction to obtain the mouse gastric cancer spontaneous tumor model. Wherein the drug contains sodium nitrite and ethyl sarcosinate, and / or the drug contains an N-nitroso compound. According to the construction method of the mouse gastric cancer spontaneous tumor model, the construction of the C57 mouse gastric cancer spontaneous tumor model by using the NESS is proposed for the first time, and the tumor formation rate can reach 75% or above; the tumor formation period is short and is about 3-4 months, and the construction cost is low.

Description

Technical Field

[0001] The present application relates to the technical field of animal model construction, and in particular to a method for constructing and applying a spontaneous gastric cancer model in mice. Background Art

[0002] Gastric cancer ranks among the top five in the world in terms of morbidity and mortality, and is characterized by late stage and heavy tumor burden. The specific pathogenesis of gastric cancer has not yet been elucidated, and there is a lack of effective treatment methods. The 5-year survival rate of advanced gastric cancer is only 30%.

[0003] Gastric cancer mouse models are the basis for the etiology of gastric cancer. Currently, mouse models in the field of gastric cancer are divided into spontaneous tumor and transplanted tumor models. The transplanted tumor model uses subcutaneous or orthotopic transplantation of human or mouse tumor cells or tissue blocks. Mouse cell lines have the advantages of high tumor formation rate and a wide range of tumor-bearing mouse strains. However, due to the lack of mature, available, and stable tumor-forming mouse gastric cancer cell lines, the commonly used mouse gastric cancer cell line is only the MFC cell line established in 1985, which has been passaged for more than hundreds of generations and is derived from the forestomach squamous cell carcinoma of 615 mice. It can only be tumor-bearing on the 615 mouse strain, and its feasibility and applicability are limited. Therefore, the current mouse gastric cancer transplanted tumor model mostly uses immunodeficient mice to implant human gastric cancer cells or tissues. However, more and more studies have shown that the tumor immune microenvironment plays an important role in the occurrence and development of tumors, which is inseparable from the development and application of tumor treatment, especially for gastric cancer, which is recognized as one of the most heterogeneous tumors. Since the stomach is a digestive organ with complex tissue structure and diverse physiological functions, there is interaction between the internal and external environments. The occurrence and development of gastric cancer is attributed to the combined effects of many environmental and genetic factors. For example, Helicobacter pylori infection is one of the most important strong risk factors for gastric cancer found so far. Therefore, the tumor immune microenvironment of gastric cancer is also one of the current research focuses in the field of gastric cancer. The gastric cancer transplant tumor model is not a primary tumor and cannot reflect the process of tumor formation in the body. It cannot simulate the tumor microenvironment. Immunodeficient mice cannot simulate the body's immune system, making it difficult to conduct research on the tumor microenvironment and immunity of gastric cancer. The use of this model for etiological research on gastric cancer is also greatly limited and controversial.

[0004] The spontaneous tumor model of mice can simulate the occurrence and development of gastric cancer in the human body, has a tumor microenvironment, and has the body's immune system. Therefore, it is widely recognized as a reliable material for the etiology of gastric cancer. Gastric cancer mouse models are roughly divided into two categories according to different pathogenic mechanisms. One is a transgenic mouse model constructed based on genetic mutations, and the other is an in situ induced mouse model constructed based on environmental stimuli. However, the current mouse spontaneous tumor model of gastric cancer also faces many challenges. The gene-edited transgenic mouse model has high specificity and has unique advantages in the study of the molecular mechanism of signaling pathways related to the occurrence and development of gastric cancer, but the production cost is high and the tumor formation cycle is long; the in situ induced mouse model constructed based on environmental stimuli has a low tumor formation rate and a long tumor formation cycle. Therefore, it is necessary to develop a new method for establishing a spontaneous gastric cancer model in mice. Summary of the invention

[0005] The purpose of this application is to provide a method for constructing a mouse gastric cancer spontaneous tumor model and its application, so as to improve the tumor formation rate in the process of constructing a mouse gastric cancer spontaneous tumor model, shorten the tumor formation cycle, and reduce the construction cost. The specific technical scheme is as follows:

[0006] The first aspect of the present application provides a method for constructing a mouse spontaneous tumor model of gastric cancer, comprising the steps of: reducing or inhibiting the expression level of the TP53 gene and / or the activity of the P53 protein in the mouse, and then administering a drug to the mouse for chemical induction to obtain a mouse spontaneous tumor model of gastric cancer; wherein the drug comprises sodium nitrite and ethyl sarcosine, and / or the drug comprises an N-nitroso compound.

[0007] In some embodiments of the present application, the reducing or inhibiting the expression level of TP53 gene and / or the activity of P53 protein in mice is to specifically knock out the TP53 gene in mice to obtain TP53 knockout mice; preferably, the TP53 knockout mice are TP53 knockout mice with systemic TP53 gene heterozygous deletion.

[0008] In some embodiments of the present application, the TP53 gene in the specific knockout mouse is at least one of exon 2, exon 3, exon 4, exon 5 and exon 6 in the TP53 gene in the specific knockout mouse genome.

[0009] In some embodiments of the present application, the specific knockout of the TP53 gene in the mouse includes using the CRISPR-Cas9 system to specifically knock out the TP53 gene in the recipient mouse.

[0010] In some embodiments of the present application, the N-nitroso compound comprises N-nitrososarcosine ethyl ester, and the concentration of the N-nitrososarcosine ethyl ester is 4g / 100mL-7g / 100mL; the concentration of the sodium nitrite is 2g / 100mL-3g / 100mL, and the concentration of the sarcosine ethyl ester is 5g / 100mL-10g / 100mL.

[0011] In some embodiments of the present application, the drug is administered by intragastric administration; the dosage of the drug is 5 mL / kg body weight to 10 mL / kg body weight.

[0012] In some embodiments of the present application, the frequency of administration of the drug is 1-3 times per week; the administration cycle of the drug is 4-8 weeks; the age of the mice when the drug is first administered is 5-7 weeks old; the weight of the mice when the drug is first administered is 20g-25g; and the mice are C57 mice.

[0013] In some embodiments of the present application, the administering of drugs to mice specifically comprises the steps of: administering the drug to mice after the mice have fasted for 2 to 4 hours; providing normal diet and water during the dosing interval; and feeding mice conventionally for 8 to 24 weeks after the administration.

[0014] The second aspect of the present application provides a mouse spontaneous gastric cancer tumor model constructed by the method described in the first aspect of the present application.

[0015] The third aspect of the present application provides the use of the method described in the first aspect of the present application or the mouse spontaneous gastric cancer tumor model described in the second aspect of the present application in screening drugs for preventing and / or treating gastric cancer.

[0016] The method for constructing a spontaneous gastric cancer model in mice provided in this application has the following advantages:

[0017] (1) The induction process is convenient. Compared with the traditional long-term administration of MNU (N-methyl-N-nitrosourea) in drinking water away from light, this method is administered intragastrically 1-3 times a week, and the operation process is more convenient;

[0018] (2) The induction process was stable, and the differences between groups were small. Compared with drinking water administration, the differences in time and dosage of oral administration were smaller.

[0019] (3) High tumor formation rate. The tumor formation rate induced by drinking water with traditional MNU compounds is about 30%, while the tumor formation rate of this method is as high as 75% or more;

[0020] (4) The tumor formation period is short. The tumor formation period of traditional methods is about 7-12 months, while that of this method is about 3-4 months.

[0021] (5) Easy to carry out. This method does not require light-proof treatment or equipment such as light-proof water bottles, and is simple to operate.

[0022] Of course, implementing any product or method of the present application does not necessarily require achieving all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application, and for ordinary technicians in this field, other embodiments can also be obtained based on these drawings.

[0024] Figure 1 The schematic diagram and renderings of the construction of the mouse gastric cancer spontaneous tumor model in Example 1 of the present application; wherein: A) is a schematic diagram of the construction of the mouse gastric cancer spontaneous tumor model, B) is a rendering of the appearance of the constructed mouse gastric cancer spontaneous tumor model; C) is a rendering of the pathological H&E staining results of the primary gastric cancer tissue of the constructed mouse gastric cancer spontaneous tumor model;

[0025] Figure 2 This is a picture showing the pathological H&E staining result of the primary gastric cancer tissue of a WT mouse that was not genetically modified in Comparative Example 2 of the present application;

[0026] Figure 3 This is the result of nuclear magnetic resonance examination of mouse stomach tissue block;

[0027] Figure 4 This is a diagram showing the effect of transplanting primary gastric cancer tissue fragments of the mouse gastric cancer spontaneous tumor model obtained in Example 1 of the present application into the subcutaneous tissue of C57BL / 6 mice. DETAILED DESCRIPTION

[0028] The following will be combined with the embodiments of the present application and the accompanying drawings to clearly and completely describe the technical solutions in the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. All other embodiments obtained by those skilled in the art based on the present application belong to the scope of protection of the present application.

[0029] In the establishment of a mouse spontaneous gastric cancer model, the tumor formation rate of a single gene mutation mouse model can reach 100%, such as INS-GAS transgenic mice, but the tumor formation period is relatively long, about 20 months; multiple gene mutations have a high tumor formation rate and a short period, but the technical requirements are high. In situ induced models include tumor formation induced by nitroso compounds or Helicobacter pylori. MNU induction has been shown to have a relatively stable tumor formation rate in current studies, but the traditional MNU induction method is likely to produce forestomach squamous cell carcinoma, with a tumor formation rate of about 30% in C57BL / 6 mice and a tumor formation period of about 7-12 months; long-term infection with Helicobacter pylori usually induces severe gastritis, and the tumor type is mostly adenocarcinoma, but the tumor formation rate that progresses to cancer is low, with a tumor formation rate of about 20% in C57BL / 6 mice, and a tumor formation period of more than 18 months. During the research process, the inventors of the present application were pleasantly surprised to find that the spontaneous gastric cancer model of C57 mice based on TP53 gene knockout has a high tumor formation rate and a short tumor formation time, which can effectively solve the research problem of the lack of TP53 knockout / mutant C57 mouse gastric cancer model.

[0030] The first aspect of the present application provides a method for constructing a mouse spontaneous tumor model of gastric cancer, comprising the steps of: reducing or inhibiting the expression level of TP53 gene and / or the activity of P53 protein in the mouse body, and then administering drugs to the mouse for chemical induction to obtain a mouse spontaneous tumor model of gastric cancer; wherein the drug contains sodium nitrite and ethyl sarcosine; and / or the drug contains an N-nitroso compound.

[0031] The genomic sequence of the mouse TP53 gene described in the present application is the nucleotide sequence of positions 69471174-69482699 of GenBank Accession No.NC_000077.7 (Update Date 2024-02), which includes 12 exons.

[0032] The P53 protein encoded by the mouse TP53 gene described in the present application is a protein whose amino acid sequence is shown in GenBank Accession No. NP_035770.2 (Update Date 2024-02).

[0033] The medicine described in the present application may also contain water, preferably double distilled water.

[0034] This application proposes for the first time to use NESS to construct a C57 mouse spontaneous gastric cancer model. The method for constructing a mouse gastric cancer spontaneous tumor model provided in this application has a high tumor formation rate. The tumor formation rate induced by drinking water with traditional MNU compounds is about 30%, while the tumor formation rate of this method can reach 75% or more; the tumor formation period is short. The tumor formation period of the traditional method is about 7-12 months, while the tumor formation period of this method is about 3-4 months, and the construction cost is low.

[0035] In some embodiments of the present application, the reducing or inhibiting the expression level of TP53 gene and / or the activity of P53 protein in mice is to specifically knock out the TP53 gene in mice to obtain TP53 knockout mice; preferably, the TP53 knockout mice are TP53 knockout mice with systemic TP53 gene heterozygous deletion.

[0036] The gene knockout mice described in this application refer to the phenomenon of inactivating a specific target gene by homologous recombination. Gene knockout is the inactivation of a specific target gene by changing the DNA sequence. This application can utilize homologous recombination of genes in mouse embryonic stem cells to heterozygously knock out the TP53 gene in the mouse genome to obtain a TP53 gene knockout mouse with systemic TP53 gene heterozygosity loss. This application does not particularly limit the specific operating steps for obtaining TP53 gene knockout mice, and the operating methods commonly used in the art can be used.

[0037] In some embodiments of the present application, the TP53 gene in the specific knockout mouse is at least one of exon 2, exon 3, exon 4, exon 5 and exon 6 in the TP53 gene in the specific knockout mouse genome.

[0038] In some embodiments of the present application, the specific knockout of the TP53 gene in the mouse includes using the CRISPR-Cas9 system to specifically knock out the TP53 gene in the recipient mouse.

[0039] In some embodiments of the present application, the N-nitroso compound comprises N-nitrososarcosine ethyl ester (NSEE), and the concentration of the N-nitrososarcosine ethyl ester is 4g / 100mL-7g / 100mL. In the present application, the concentration of N-nitrososarcosine ethyl ester can be 4g / 100mL, 4.5g / 100mL, 5g / 100mL, 5.5g / 100mL, 6g / 100mL, 6.5g / 100mL, 7g / 100mL, or a range consisting of any two values ​​therebetween. The N-nitrososarcosine ethyl ester of the present application can be prepared from sodium nitrite (NaNO2) and sarcosine ethyl ester.

[0040] In some embodiments of the present application, the concentration of the sodium nitrite is 2g / 100mL-3g / 100mL, and the concentration of the sarcosine ethyl ester is 5g / 100mL-10g / 100mL. In the present application, the concentration of sodium nitrite can be 2g / 100mL, 2.5g / 100mL, 3g / 100mL, or a range consisting of any two values ​​therebetween; the concentration of the sarcosine ethyl ester can be 5g / 100mL, 6g / 100mL, 7g / 100mL, 8g / 100mL, 9g / 100mL, 10g / 100mL, or a range consisting of any two values ​​therebetween.

[0041] In some embodiments of the present application, the drug is administered by gavage; the dosage of the drug is 5 mL / kg body weight-10 mL / kg body weight (referring to the body weight of the mouse). In the present application, the dosage of the drug can be 5 mL / kg body weight, 6 mL / kg body weight, 7 mL / kg body weight, 8 mL / kg body weight, 9 mL / kg body weight, 10 mL / kg body weight or a range consisting of any two values ​​therebetween. In the present application, the dosage of the drug is about 200 µL / mouse-250 µL / mouse.

[0042] In some embodiments of the present application, the frequency of administration of the drug is 1-3 times per week; the administration cycle of the drug is 4-8 weeks; the age of the mouse when the drug is first administered is 5-7 weeks old; the weight of the mouse when the drug is first administered is 20g-25g; the mouse is a C57 mouse. The C57 mouse of the present application can be a C57BL / 6 mouse. In the present application, the frequency of administration of the drug can be 1, 2 or 3 times per week; the administration cycle of the drug can be 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks; the age of the mouse when the drug is first administered can be 5 weeks, 5.5 weeks, 6 weeks, 6.5 weeks or 7 weeks old; the weight of the mouse when the drug is first administered can be 20g, 21g, 22g, 23g, 24g, 25g or a range consisting of any two values ​​therebetween.

[0043] In some embodiments of the present application, the administration of drugs to mice specifically includes the steps of: administering drugs to mice after fasting for 2 to 4 hours; giving normal diet and drinking water during the dosing interval; and conventionally raising mice for 8 to 24 weeks after the administration. In the present application, the fasting time of mice can be 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, or a range consisting of any two values ​​therebetween; and the conventional raising time after the administration can be 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, 22 weeks, 24 weeks, or a range consisting of any two values ​​therebetween.

[0044] The method for constructing a spontaneous tumor model of gastric cancer in mice provided in the present application has a convenient induction process. Compared with the traditional MNU long-term administration of drinking water in the dark, this method has fewer gavages per week and a more convenient operation process. The induction process is stable and the differences between the groups are small. Compared with the administration of drinking water, the differences in the time and amount of gavage administration are smaller. The method is easy to carry out, does not require light protection treatment, does not require equipment such as light-shielding water bottles, and is simple to operate.

[0045] The second aspect of the present application provides a mouse spontaneous gastric cancer tumor model constructed by the method described in the first aspect of the present application.

[0046] The third aspect of the present application provides the use of the method described in the first aspect of the present application or the mouse spontaneous gastric cancer tumor model described in the second aspect of the present application in screening drugs for preventing and / or treating gastric cancer.

[0047] Example

[0048] Hereinafter, the embodiments of the present application will be described in more detail with reference to Examples and Comparative Examples. Various tests and evaluations were performed according to the following methods. In addition, unless otherwise specified, "parts" and "%" are by mass.

[0049] Example 1

[0050] (1) The CRISPR-Cas9 system was used to knock out exon 2, exon 3, exon 4, exon 5, and exon 6 of the TP53 gene in the C57BL / 6 mouse genome. After PCR identification, TP53 gene knockout C57BL / 6 mice with systemic TP53 gene heterozygous loss (P53 - / + ).

[0051] (2) Thirty 6-week-old C57BL / 6 mice with systemic TP53 gene heterozygous deletion and body weight of 20 g-25 g were selected and fasted for 3 hours. The C57BL / 6 mice were gavaged with drugs, including sodium nitrite at a concentration of 3 g / 100 mL and sarcosine ethyl ester at a concentration of 10 g / 100 mL, and the solvent was double distilled water. The dosage of the drug was 10 mL / kg body weight, and the administration frequency was twice a week. Normal diet and drinking water were given between administrations for a total of 6 weeks. After the end of the administration, the mice were fed normally for 16 weeks.

[0052] Comparative Example 1

[0053] (1) The CRISPR-Cas9 system was used to knock out exon 2, exon 3, exon 4, exon 5, and exon 6 of the TP53 gene in the C57BL / 6 mouse genome. After PCR identification, TP53 gene knockout C57BL / 6 mice with systemic TP53 gene heterozygous loss (P53- / + ).

[0054] (2) Six 6-week-old C57BL / 6 mice with heterozygous deletion of the TP53 gene and body weight of 20 g to 25 g were selected. After fasting for 3 hours, the C57BL / 6 mice were gavaged with PBS solution twice a week. The mice were given normal diet and water during the dosing intervals for a total of 6 weeks. After the end of the dosing period, they were kept in a conventional manner for 16 weeks.

[0055] Comparative Example 2

[0056] Fifteen 6-week-old non-genetically modified C57BL / 6 mice (WT) weighing 20g-25g were selected. After fasting for 3 hours, the C57BL / 6 mice were gavaged with drugs, including sodium nitrite with a concentration of 3g / 100mL and sarcosine ethyl ester with a concentration of 10g / 100mL, and the solvent was double distilled water; the drug dosage was 10mL / kg body weight, the administration frequency was 2 times a week, and normal diet and drinking water were given between administrations, and the total administration lasted for 6 weeks; after the end of the administration, the mice were fed normally for 16 weeks.

[0057] The mice finally obtained in the above examples and comparative examples were subjected to tissue sampling and histopathological (H&E staining) identification. The specific results are shown in Tables 1 and Figure 1-Figure 3 shown.

[0058] Table 1

[0059]

[0060] From the above results, it can be seen that Example 1 obtained 23 mouse gastric cancer spontaneous tumor models, with a tumor formation rate of 76.6% (23 / 30), and a tumor formation period of about 3 months after oral administration. However, the spontaneous gastric cancer model of mice obtained in Comparative Example 2 was only 6.7% (1 / 15), and Comparative Example 1 failed to successfully obtain a mouse gastric cancer spontaneous tumor model.

[0061] The pathological results of the mouse gastric cancer spontaneous tumor model obtained in Example 1 of the present application showed that the pathological types of gastric cancer that occurred were gastric adenocarcinoma and gastric squamous cell carcinoma. Figure 1 The schematic diagram and renderings of the construction of the mouse spontaneous gastric cancer tumor model in Example 1 of the present application; wherein: A) is a schematic diagram of the construction of the mouse spontaneous gastric cancer tumor model, B) is a rendering of the appearance of the constructed mouse spontaneous gastric cancer tumor model; C) is a pathological H&E staining result of the primary gastric cancer tissue of the constructed mouse spontaneous gastric cancer tumor model.

[0062] Figure 2 This is a graph showing the pathological H&E staining results of primary gastric cancer tissue of a WT mouse that was not genetically modified in Comparative Example 2 of the present application.

[0063] Figure 3Figure 1 is a graph showing the results of nuclear magnetic resonance (NMR) detection of gastric tissue blocks of mice; wherein: A is a graph showing the results of nuclear magnetic resonance (NMR) detection of gastric tissue blocks of WT mice (not suffering from gastric cancer) that have not been genetically modified in Comparative Example 2, and B is a graph showing the results of nuclear magnetic resonance (NMR) detection of gastric tissue blocks of the spontaneous gastric cancer model of mice obtained in Example 1; Figure 3 It can be seen that the stomach wall of the gastric tissue block of the mouse gastric cancer spontaneous tumor model obtained in Example 1 is thickened, and its thickness is significantly greater than the thickness of the gastric tissue block of the non-genetically modified WT mouse (not suffering from gastric cancer) in Comparative Example 2.

[0064] The primary gastric cancer tissue fragments of the mouse gastric cancer spontaneous tumor model obtained in Example 1 of the present application are transplanted subcutaneously into C57BL / 6 mice to construct an allogeneic transplantation model. Figure 4 This is a diagram showing the effect of transplanting the primary gastric cancer tissue fragments of the mouse gastric cancer spontaneous tumor model obtained in Example 1 of the present application into the subcutaneous tissue of C57BL / 6 mice. Figure 4 It can be seen that the allogeneic transplantation model was successfully established.

[0065] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. A method for constructing a mouse spontaneous gastric cancer model, comprising the steps of: reducing or inhibiting the expression of TP53 gene and / or the activity of P53 protein in the mouse, and then administering drugs to the mouse for chemical induction to obtain a mouse spontaneous gastric cancer model; wherein: The drug comprises sodium nitrite and ethyl sarcosinate, and / or the drug comprises an N-nitroso compound.

2. The construction method according to claim 1, wherein: The reducing or inhibiting of the expression level of TP53 gene and / or the activity of P53 protein in mice is to specifically knock out the TP53 gene in mice to obtain TP53 gene knockout mice; preferably, the TP53 gene knockout mice are TP53 gene knockout mice with systemic TP53 gene heterozygous deletion.

3. The construction method according to claim 2, wherein: The TP53 gene in the specific knockout mouse is at least one of the 2nd exon, the 3rd exon, the 4th exon, the 5th exon and the 6th exon in the TP53 gene in the genome of the specific knockout mouse.

4. The construction method according to claim 2, wherein: The specific knockout of the TP53 gene in the mouse includes using the CRISPR-Cas9 system to specifically knock out the TP53 gene in the recipient mouse.

5. The construction method according to claim 1, wherein: The N-nitroso compound comprises N-nitrososarcosine ethyl ester, and the concentration of the N-nitrososarcosine ethyl ester is 4 g / 100 mL-7 g / 100 mL; The concentration of the sodium nitrite is 2g / 100mL-3g / 100mL, and the concentration of the sarcosine ethyl ester is 5g / 100mL-10g / 100mL.

6. The construction method according to claim 1, wherein: The drug is administered by intragastric administration; the dosage of the drug is 5 mL / kg body weight to 10 mL / kg body weight.

7. The construction method according to claim 1, wherein: The drug is administered once to three times a week for a period of 4 to 8 weeks. The age of the mice when the drug is first administered is 5 to 7 weeks old. The weight of the mice when the drug is first administered is 20 g to 25 g. The mice are C57 mice.

8. The construction method according to any one of claims 1 to 7, wherein: The specific steps of administering the drug to mice include: administering the drug after the mice have fasted for 2 to 4 hours; providing normal diet and water during the administration interval; and feeding the mice conventionally for 8 to 24 weeks after the administration.

9. A mouse spontaneous gastric cancer tumor model constructed by the method according to any one of claims 1 to 8.

10. Use of the method according to any one of claims 1 to 8 or the mouse spontaneous gastric cancer tumor model according to claim 9 in screening drugs for preventing and / or treating gastric cancer.

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