Construction method and application of drosophila m6A RNA methylation tumor model

Abstract

The invention discloses a construction method and application of a drosophila m6A RNA methylation tumor model. Drosophila melanogaster is used as a model organism, an m6A related gene is specifically overexpressed in a drosophila melanogaster eye adult disc through an MARCM system, and the m6A RNA methylation tumor model is constructed. The method ensures the accuracy and reliability of the model, and provides a reliable in-vivo tumor model for researching the effect of m6A in tumor generation and development. By utilizing the characteristics of high visibility and non-invasiveness of the drosophila melanogaster model, the blank of m6A methylation in drosophila melanogaster tumor research is filled. The model constructed by the invention can more accurately simulate the effect of m6A methylation in in-vivo tumors through specific expression of m6A related genes and tumor induction factors, and provides an important in-vivo experimental tool for studying the molecular mechanism of m6A in tumor biology.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical research, and particularly relates to a method for constructing a Drosophila m6A RNA methylation tumor model and its application. This model aims to systematically screen and verify the key molecular mechanisms regulating m6A methylation in tumorigenesis and development through in vivo experiments in Drosophila, providing an important experimental tool for deeply understanding the role of m6A methylation in tumor biology. Background Art

[0002] Drosophila melanogaster, as a classical model organism, has extensive applications in genetics and developmental biology research due to its unique biological advantages. Drosophila has characteristics such as a short life cycle, strong reproductive ability, simple genome, and easy operation, making it an ideal model for studying gene function and complex biological processes. In addition, many genes and signaling pathways in Drosophila are highly conserved with humans, which makes the biological discoveries obtained in Drosophila often have important reference value.

[0003] Drosophila also has unique advantages in constructing tumor models. Through gene editing techniques, such as the MARCM (Mosaic Analysis with a Repressible Cell Marker) system, specific genes can be overexpressed or knocked out in specific tissues of Drosophila, thus simulating the occurrence and development of human tumors. For example, the Drosophila eye imaginal disc is a commonly used model system. By overexpressing oncogenes (Ras and Raf activators) in the eye imaginal disc, the formation and growth of tumors can be observed, providing an intuitive experimental platform for studying the molecular mechanisms of tumorigenesis. The scribble gene encodes a cell polarity protein that participates in maintaining cell polarity and tissue structural integrity. Drosophila with scribble gene mutations show disrupted cell polarity and damaged tissue structure, and these effects are similar to the loss of cell polarity in human tumors. Introducing mutations in the scribble gene into cells overexpressing oncogenes (Ras and Raf activators) will lead to the loss of cell polarity, thus promoting tumor formation. This model can not only simulate the tumor growth process, but also directly observe the size and morphology of tumors through a microscope, providing a highly visible and non-invasive research method.

[0004] RNA methylation, especially N6-methyladenosine (m6A) modification, has been a hot topic of research in recent years. m6A modification is widely involved in various biological processes by regulating RNA transcription, maturation, translation, degradation and stability. In in vitro studies, m6A modification has been shown to be closely related to the occurrence and development of various cancers. For example, abnormal expression of m6A modifying enzymes (such as METTL3, METTL14) and demethylases (such as FTO, ALKBH5) can affect the proliferation, metabolism, immune response and microenvironment formation of tumor cells. However, the limitation of in vitro studies is that it is difficult to fully reflect the complex physiological and pathological environment in vivo. Therefore, in vivo studies are particularly important.

[0005] In vivo studies, the role of RNA methylation has also been gradually revealed. Through studies in mouse models, m6A modification has been shown to play a key role in the occurrence and development of tumors. For example, m6A modification can regulate the expression of tumor-related genes and affect the proliferation and metastasis of tumor cells. However, these studies are often limited by high costs, complex operations, and long experimental cycles, which makes fruit flies a more ideal model organism.

[0006] Although there are relatively few studies on RNA methylation in Drosophila, some preliminary findings have been made. The m6A RNA methyltransferase also exists in Drosophila and plays an important role in the development of Drosophila. The m6A methylation regulatory factors in Drosophila are highly homologous to those in mammals, which provides a unique advantage for studying m6A methylation using the Drosophila model. For example, the m6A methyltransferase complex mettl3 / mettl14 / nito in Drosophila corresponds to METTL3 / METTL14 / RBM15 in mammals, and these proteins are highly similar in structure and function. By constructing a Drosophila m6A RNA methylation tumor model, the role of m6A modification in tumor occurrence and development can be studied more systematically.

[0007] However, the specific mechanism and function of Drosophila m6A RNA methylation in tumors in vivo are still unclear. The existing methods for constructing tumor models in mice are complex and difficult to efficiently, quickly and accurately simulate the role of m6A in tumor development. These limitations seriously hinder the in-depth study of m6A methylation in tumors in vivo. Therefore, the present invention aims to develop an efficient and accurate method for constructing a Drosophila m6A RNA methylation tumor model to overcome the shortcomings of the prior art. Summary of the invention

[0008] The problem to be solved in this application is how to construct an m6A RNA methylation tumor model in Drosophila: This application uses Drosophila as a model organism, and specifically overexpresses the m6A-related gene nito (a homolog of the human m6A RNA methylation regulatory factor RBM15) in the imaginal disc of the Drosophila eye through the MARCM system to construct an m6A RNA methylation tumor model. This method ensures the accuracy and reliability of the model and provides a reliable in vivo tumor model for studying the role of m6A in tumor development. This application uses the high visibility and non-invasive characteristics of the Drosophila model to fill the gap in m6A methylation research in Drosophila tumors. After the model is constructed, the specific role of the screened genes in tumor development can be confirmed through high-throughput mutation screening and systematic functional verification experiments, providing new perspectives and tools for a deep understanding of the function of m6A in tumors. This model can more accurately simulate the role of m6A methylation in in vivo tumors by specifically expressing m6A-related genes and tumor-inducing factors, and provides an important in vivo experimental tool for studying the molecular mechanism of m6A in tumor biology.

[0009] The present invention is implemented by the following technical solutions:

[0010] A method for constructing a Drosophila m6A RNA methylation tumor model specifically comprises the following steps:

[0011] ey-FLP1; actin>y + >Gal4,UAS-GFP;FRT82B,Tubulin-Gal80 Drosophila strains and UAS-nito;UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb Drosophila strain hybridization; construct the Drosophila m6ARNA methylation tumor model in the eye imaginal disc, and select the genotype ey-FLP1; actin>y + >Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,scrib - / - You can get the fruit flies.

[0012] The present invention uses MARCM technology to overexpress Raf in Drosophila eye imaginal disc cells GOF The polarity-related gene scribble was mutated and the RNA methylase nito was overexpressed, thereby constructing a tumor model with high expression of m6A RNA methylation in the imaginal disc of the Drosophila eye. Compared with the closest prior art, the biggest difference of this application is that through specific gene manipulation and a combination of multiple steps, an efficient m6A RNA methylation tumor model is constructed.

[0013] First, the present invention uses the MARCM system to overexpress Raf in Drosophila eye imaginal disc cells. GOF Gene. Raf GOF Gene overexpression is a key step in building a tumor model. Through this operation, tumor formation can be induced in the imaginal disc of the Drosophila eye. This technical approach ensures the reliability and stability of the tumor model.

[0014] Secondly, the present application enhances tumor formation by mutating the polarity-related gene scribble. Mutation of the scribble gene leads to loss of cell polarity, further promoting tumor growth. This step not only increases the complexity and authenticity of the tumor model, but also provides a new perspective for studying the role of cell polarity in tumorigenesis.

[0015] Finally, the present application simultaneously overexpresses the RNA methylase nito on the basis of the above. Overexpression of nito can significantly increase the level of m6A RNA methylation in the imaginal disc of the Drosophila eye, and the tumor model with high expression of m6A RNA has stronger proliferation and invasion capabilities. This technical means is the core innovation of the present application. Through the coordinated operation of multiple genes, it not only successfully constructs an efficient m6A RNA methylation tumor model, but also provides a new tool for studying the role of m6A methylation in tumor occurrence and development.

[0016] In summary, the biggest difference of this application is that by adding and changing multiple key steps in the manufacturing method, including specific overexpression of Raf GOF The researchers used a combination of specific gene manipulations and multiple steps to construct an efficient m6A RNA methylation tumor model, by combining the m6A RNA methylation gene, the scribble gene, and the overexpression of the RNA methylase nito. These specific gene manipulations and multiple steps not only ensure the accuracy and reliability of the model, but also fill the gap in m6A methylation research in Drosophila tumors.

[0017] The inventive principle of the present invention is:

[0018] The MARCM system was used to specifically overexpress m6A-related genes (such as METTL3, METTL14, WTAP, etc.) and the key kinase Raf in the Ras / MAPK signaling pathway in the Drosophila eye imaginal disc through UAS / GAL4, and then the homozygous mutation of the polarity-related gene scribble was introduced during the cell mitosis period using the FLP / FRT system to construct a Drosophila in vivo tumor model with high expression of m6A RNA. Overexpression of m6A-related genes can promote the proliferation and growth of tumor cells, thereby forming an obvious tumor phenotype. Taking advantage of the advantages of Drosophila, such as high visibility, simple operation and low cost, the key factors regulating m6A methylation can be screened and verified more efficiently, providing new perspectives and tools for a deeper understanding of the function of m6A in tumors, filling the gap in m6A methylation research in Drosophila tumors.

[0019] The beneficial effects of the present invention are:

[0020] This paper uses the MARCM system for the first time to specifically overexpress m6A-related genes in Drosophila eye imaginal discs, and successfully constructs a Drosophila tumor model with high expression of m6A RNA methylation. This precise gene regulation method ensures the accuracy and reliability of the model and provides a reliable platform for studying the role of m6A in tumor occurrence and development.

[0021] The Drosophila m6A RNA methylation tumor model constructed by the present invention can be used to verify potential drug targets and accelerate the early stages of drug discovery. The model can be used to evaluate the effects of candidate drugs in vivo and improve the success rate and efficiency of drug development. By screening and verifying m6A methylation-related genes, new biomarkers can be discovered for early diagnosis and prognosis evaluation of tumors. Understanding the specific role of m6A methylation in tumors will help develop personalized treatment plans and improve treatment effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The figure is a schematic diagram of the MARCM system. The system combines the FLP / FRT recombination system with the UAS / Gal4 system, using the FLP recombinase to cut the chromosome fragment between the FRT sites at a specific time point, remove the Gal80 repressor, and allow the Gal4-driven UAS gene to be expressed in specific cells, thereby achieving precise regulation and labeling of the target gene.

[0023] Figure 2 UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - Flowchart for the construction of the / SM6B-TM6B.Tb Drosophila strain.

[0024] Figure 3Flowchart for using the MARCM system in Drosophila to simultaneously express activated Raf, nito and mutate the scribble gene in cells. During mitosis, under the action of the FLP / FRT elements, chromosomes are recombined to form cells with three genotypes.

[0025] Figure 4 Flow chart for constructing m6A methylated tumors in Drosophila. The m6A methylated tumor model was constructed in the imaginal disc of Drosophila eyes using the MARCM system. The GFP cells in the figure represent tumor cells. The larger the GFP area, the stronger the tumor proliferation ability. DETAILED DESCRIPTION

[0026] The present invention adopts the method of genetic hybridization and recombination of Drosophila to overexpress Raf in Drosophila eye imaginal disc by Gal4 / UAS system and MARCM system. GOF By mutating scribble and overexpressing the RNA methylase nito, tumors with high expression of m6A RNA methylation were induced in the eye imaginal disc. This model can be used to study the role of m6A methylation in tumor development and is a novel in vivo tumor model for studying m6A methylation regulatory signals.

[0027] The MARCM system is a powerful tool for cell-specific gene manipulation in Drosophila. The system uses the FLP / FRT recombination system and the UAS / Gal4 system to activate or inhibit genes in specific cells. Specifically, the MARCM system uses the FLP recombinase to cut the chromosome segment between the FRT sites at a specific time point, thereby removing the Gal80 repressor and allowing the Gal4-driven UAS gene to be expressed. In this way, precise regulation of target genes in specific cells or tissues can be achieved (such as Figure 1 ).

[0028] N6-methyladenosine (m6A) is one of the most common modifications in RNA. It is widely involved in various biological processes by regulating RNA transcription, maturation, translation, degradation and stability. Nito is an RNA methylase, a homolog of human RBM15, which can catalyze the formation of m6A modification. In Drosophila, overexpression of nito can significantly increase RNA methylation levels, thereby affecting gene expression and cell function. Therefore, by overexpressing nito in Drosophila, a model of high expression of m6A RNA methylation can be constructed to study the role of m6A modification in tumor development.

[0029] Fruit flies are usually raised in standard fruit fly tubes or fruit fly bottles, with corn culture medium at the bottom and latex sponge plugs at the top to ensure air permeability. The culture medium is prepared by mixing corn flour, yeast, sugar and water, and sometimes a small amount of acetic acid is added to prevent mold growth. Fruit flies grow best in a constant temperature environment of about 25°C, and the life cycle of each generation is about 10-14 days.

[0030] In order to construct an efficient m6A RNA methylation tumor model, we used two key fruit fly strains and achieved precise gene regulation through hybridization. Fruit flies have four chromosomes, and the present invention mainly performs hybridization and recombination on chromosomes 2, 3 and X.

[0031] The tools fruit flies need to use include:

[0032] a.ey-FLP1; Sp / CyO; Sb / TM6B.Tb: The X chromosome carries the recombinase FLP1 specifically expressed in the eye imaginal disc, and chromosomes 2 and 3 carry balancers. The fruit fly has hairy axillae, curled wings, hairy shoulders, and truncated hair.

[0033] b.FRT82B, located on chromosome 3, has white eyes, carries the neo resistance gene, and is a tool fruit fly used for recombination.

[0034] c.Sp / CyO;Sb / TM6B.Tb and sco / CyO;Sb / TM6B.Tb were tools used for balancing Drosophila.

[0035] d.Sp; Sb / SM6B-TM6B.Tb is a genetically coupled tool fly, which carries balancers on chromosomes 2 and 3, showing axillary hairiness (Sb), curly wings (CyO) and shoulder hairiness and truncated hair (TM6B.Tb). The two balancer chromosomes SM6B-TM6B.Tb are linked together and inherited, ensuring the stability of the genotype during hybridization.

[0036] In this project, we need to construct the following two genotypes of fruit flies for experiments:

[0037] 1. Drosophila strain 1: ey-FLP1; actin>y + >Gal4, UAS-GFP; FRT82B, Tubulin-Gal80

[0038] ey-FLP1 (X chromosome): ey-FLP1 is a strain that expresses FLP recombinase specifically in the eye imaginal disc and is used to initiate FLP / FRT recombination during mitosis.

[0039] actin>y +>Gal4 (Chromosome II): This is a universal Gal4 driver system used to activate the expression of UAS genes in specific cells.

[0040] UAS-GFP (chromosome II): UAS-GFP is a reporter gene used to mark cells expressing Gal4 for easy observation and screening.

[0041] FRT82B, Tubulin-Gal80 (chromosome III): Tubulin-Gal80 is a ubiquitously expressed Gal80 repressor used to inhibit Gal4-driven UAS gene expression prior to FLP recombination.

[0042] The method for constructing Drosophila strain 1 is as follows:

[0043] First, actin>y + >Gal4 Drosophila strain was crossed with UAS-GFP Drosophila strain to obtain actin>y + >Gal4 / UAS-GFP female virgin fruit flies were then crossed with Sp / CyO; Sb / TM6B.Tb male fruit flies for two generations to obtain actin>y + >Gal4, UAS-GFP / CyO; Sb / TM6B.Tb.

[0044] Subsequently, the FRT82B fruit fly strain was hybridized with the Tubulin-Gal80 fruit fly strain to obtain FRT82B / Tubulin-Gal80 female virgin fruit flies, which were then hybridized with Sp / CyO; Sb / TM6B.Tb male fruit flies to obtain the Sp / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb fruit fly strain.

[0045] Next, actin>y + >Gal4,UAS-GFP / CyO;Sb / TM6B.Tb fruit fly strain was crossed with Sp / CyO;FRT82B,Tubulin-Gal80 / TM6B.Tb fruit fly strain to construct actin>y + >Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb fruit fly strain.

[0046] Finally, ey-FLP1;Sp / CyO;Sb / TM6B.Tb male and female virgin flies were crossed with actin>y +>Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb male fruit flies were hybridized, and finally the ey-FLP1; actin>y+>Gal4, UAS-GFP; FRT82B, Tubulin-Gal80 fruit fly strain was obtained.

[0047] 2. Drosophila strain 2: UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb

[0048] UAS-nito (Chromosome II): UAS-nito is a strain expressing the nito gene under the drive of Gal4 and is used to overexpress nito in specific cells.

[0049] UAS-Raf GOF (Chromosome III): UAS-Raf GOF This is a strain that expresses the activation fragment of Raf under the drive of Gal4.

[0050] FRT82B,scrib - / - (Chromosome III): scrib - / - This is a strain in which the polarity-related gene scribble is mutated on chromosome 3 of Drosophila, which is used to simulate the loss of cell polarity.

[0051] The construction process of Drosophila strain 2 is as follows Figure 2 , the specific method is:

[0052] First, the UAS-Raf GOF ,FRT82B,scrib - / - The Drosophila strain was crossed with the sco / CyO; Sb / TM6B.Tb strain for two generations to obtain the sco / CyO; UAS-Raf GOF ,FRT82B,scrib - / - The UAS-nito / CyO;Sb / TM6B.Tb fruit fly strain was obtained by hybridizing the UAS-nito and sco / CyO;Sb / TM6B.Tb fruit fly strains for two generations.

[0053] Then sco / CyO; UAS-Raf GOF ,FRT82B,scrib - / - The UAS-nito / CyO; UAS-Raf / TM6B.Tb fruit fly strain was hybridized with the UAS-nito / CyO; Sb / TM6B.Tb fruit fly strain to obtain the UAS-nito / CyO; UAS-Raf GOF,FRT82B,scrib - / - / TM6B.Tb fruit fly strain, and then hybridized with Sp; Sb / SM6B-TM6B.Tb to select UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb genotype fruit flies.

[0054] By hybridizing the two fruit fly strains mentioned above, precise gene regulation and tumor model construction can be achieved.

[0055] 1. Hybridization process:

[0056] Virgin flies of Drosophila line 1 (ey-FLP1; actin>y + >Gal4,UAS-GFP; FRT82B,Tubulin-Gal80) and Drosophila line 2 male flies (UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb) for hybridization.

[0057] 2. Regulation of gene expression:

[0058] In the eye imaginal disc, ey-FLP1 initiates FLP recombination during cell mitosis, cuts off Tubulin-Gal80 between the FRT82B sites, removes the Gal80 repressor, allows Gal4 to be expressed freely, and causes a homozygous mutation in the scribble gene.

[0059] Gal4-driven UAS system will activate UAS-Raf GOF , UAS-nito and UAS-GFP to achieve overexpression of Raf, overexpression of nito and expression of GFP respectively.

[0060] 3. Specific genotype ( Figure 3 ):

[0061] Through hybridization, the genotypes of the offspring fruit flies were: ey-FLP1; actin>y+>Gal4, UAS-GFP / UAS-nito; FRT82B, Tubulin-Gal80 / UAS-Raf GOF ,FRT82B,scrib - / - .

[0062] During mitosis, under the action of FLP / FRT elements, chromosomes are recombined to form three genotypes of cells:

[0063] a.ey-FLP1; actin>y+ >Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,

[0064] scrib - / - The scribble in the cell was homozygous for mutation and expressed activated Raf and nito, which were labeled with GFP and emitted green fluorescence.

[0065] b.ey-FLP1; actin>y + >Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,Tubulin-Gal80 /

[0066] FRT82B, Tubulin-Gal80. In this cell, no other genes are expressed due to the presence of Tubulin-Gal80, and the cell does not have fluorescence.

[0067] c.ey-FLP1; actin>y+>Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,

[0068] Tubulin-Gal80. The scribble gene in this cell is a heterozygous mutation, and the presence of Tubulin-Gal80 means that other genes are not expressed and the cell does not fluoresce.

[0069] Through the above hybridization and gene expression regulation, a tumor model with high expression of m6A RNA methylation in the imaginal disc of the Drosophila eye was successfully constructed. The genotype selected was ey-FLP1; actin>y + >Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,scrib - / - The Drosophila m6A RNA methylation tumor model was obtained by using Drosophila. The effectiveness of the model was verified by observing and analyzing the size and morphology of the tumor.

[0070] In this model, overexpression of activated Raf and nito, and mutation of scribble increased the m6A RNA methylation level and promoted tumor proliferation and invasion (e.g. Figure 4). By marking tumor cells with GFP, it is easy to visually detect and analyze the size and morphology of the tumor, and it is possible to screen and verify the key factors that regulate m6A methylation, providing a powerful tool for in-depth research on the role of m6A in tumor occurrence and development.

Claims

1. A method for constructing a Drosophila m6A RNA methylation tumor model, characterized in that: The construction method specifically comprises the following steps: ey-FLP1; actin>y + >Gal4,UAS-GFP;FRT82B,Tubulin-Gal80 Drosophila strains and UAS-nito;UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb Drosophila strain hybridization; construct the Drosophila m6ARNA methylation tumor model in the eye imaginal disc, and select the genotype ey-FLP1; actin>y + >Gal4,UAS-GFP; UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / FRT82B,scrib - / - You can get the fruit flies.

2. The method for constructing a Drosophila m6A RNA methylation tumor model according to claim 1, characterized in that: The method for constructing the Drosophila m6A RNA methylation tumor model in the eye imaginal disc is as follows: In the eye imaginal disc, ey-FLP initiates FLP recombination during cell mitosis, cuts off Tubulin-Gal80 between FRT82B sites, removes the Gal80 repressor, allows Gal4 to be freely expressed, and the scribble gene undergoes a homozygous mutation; Gal4-driven UAS system activates UAS-Raf GOF , UAS-nito and UAS-GFP to achieve overexpression of Raf, overexpression of nito and expression of GFP respectively.

3. The method for constructing a Drosophila m6A RNA methylation tumor model according to claim 1, characterized in that: ey-FLP1; actin>y + >Gal4,UAS-GFP;FRT82B,Tubulin-Gal80 Drosophila strain was constructed by: + >Gal4,UAS-GFP / CyO;Sb / TM6B.Tb fruit fly strain was crossed with Sp / CyO;FRT82B,Tubulin-Gal80 / TM6B.Tb fruit fly strain, and the genotype actin>y + >Gal4, UAS-GFP / CyO; FRT82B, Tubulin-Gal80 / TM6B.Tb fruit fly strain, and then hybridized with ey-FLP1; Sp / CyO; Sb / TM6B.Tb fruit flies to obtain ey-FLP1; actin>y + >Gal4,UAS-GFP;FRT82B,Tubulin-Gal80 genotypes of Drosophila.

4. The method for constructing a Drosophila m6A RNA methylation tumor model according to claim 1, characterized in that: UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - The method for constructing the / SM6B-TM6B.Tb fruit fly strain is as follows: UAS-Raf GOF ,FRT82B,scrib - / - The sco / CyO; Sb / TM6B.Tb flies were crossed for two generations to construct the sco / CyO; UAS-Raf GOF ,FRT82B,scrib - / - / TM6B.Tb Drosophila strain; UAS-nito was crossed with sco / CyO; Sb / TM6B.Tb fruit flies for two generations to construct the UAS-nito / CyO; Sb / TM6B.Tb fruit fly strain. Subsequently, sco / CyO; UAS-Raf GOF ,FRT82B,scrib - / - The UAS-nito / CyO; UAS-Raf / TM6B.Tb fruit fly strain was hybridized with the UAS-nito / CyO; Sb / TM6B.Tb fruit fly strain to obtain the UAS-nito / CyO; UAS-Raf GOF ,FRT82B,scrib - / - / TM6B.Tb fruit fly strain, and then hybridized with Sp; Sb / SM6B-TM6B.Tb to obtain UAS-nito; UAS-Raf GOF ,FRT82B,scrib - / - / SM6B-TM6B.Tb Drosophila strain.

5. An application of a Drosophila m6A RNA methylation tumor model, characterized in that: The Drosophila m6A RNA methylation tumor model is constructed using the method described in any one of claims 1 to 4, and the model is used to discover new tumor biomarkers and verify potential tumor drug targets.

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