Duodenal cancer cell strain TH-1 and separation and purification method thereof

Abstract

The invention belongs to the technical field of biology, and relates to a duodenal cancer cell strain TH-1 and a separation and purification method thereof. The cell strain TH-1 contains ATM gene mutation, can efficiently express DHX33 protein, and is highly sensitive to a DHX33 helicase inhibitor. The invention also relates to a method for screening anti-cancer drugs, which comprises the following steps: contacting TH-1 cells with a compound to be detected, and judging whether the compound to be detected can be used for treating duodenal cancer with ATM gene mutation or not. The invention also relates to application of the duodenal cancer cell strain TH-1 in screening of anti-cancer drugs. The cell strain TH-1 has unique ATM gene mutation and unique STR cell identification atlas, and can be used for duodenal cancer anti-cancer drug screening and gene expression research.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to a duodenal cancer cell line with ATM gene mutation and a separation and purification method thereof. BACKGROUND

[0002] The duodenum is mostly located in the retroperitoneum and is closely related to the pancreas, so duodenal tumors, especially malignant tumors, are prone to invade the head of the pancreas. Primary duodenal tumors refer to benign or malignant tumors occurring in each segment of the duodenum, which are divided into primary duodenal benign tumors and primary duodenal malignant tumors, but do not include tumors in the Vater ampulla and the distal end of the common bile duct. In China, primary duodenal tumors account for 13%-36% of small intestinal tumors; in foreign countries, primary duodenal tumors account for 18.4%-52.0% of small intestinal tumors. Primary duodenal tumors are more common in middle-aged people and are mostly located in the descending and horizontal parts of the duodenum, and are most commonly found in the papillary part. The pathological type of benign tumors is mostly adenoma, and the pathological type of malignant tumors is mostly adenocarcinoma.

[0003] Because the duodenum has special anatomical and physiological characteristics, it lacks early specific clinical manifestations and is prone to delayed diagnosis. Common clinical symptoms include: (1) digestive system symptoms, such as abdominal pain, nausea, anorexia, and symptoms similar to digestive system inflammation and ulcer; (2) weight loss and anemia are common symptoms of duodenal tumors, especially in patients with a long medical history and advanced malignant tumors; (3) jaundice: more common in the descending part of the duodenum with large tumors, especially in tumors blocking the opening of the biliopancreatic duct in the papillary region, often accompanied by abdominal pain; (4) digestive system bleeding: fecal occult blood positivity and melena are more common, and hematemesis cases are rare; (5) nausea, vomiting, and intestinal obstruction symptoms: more common in rapidly growing tumors with large sizes, causing intestinal obstruction, or malignant tumors infiltrating and growing through the serosa; (6) abdominal mass: less common in clinical practice, mostly seen in large serosal tumors growing outward. Abdominal pain, jaundice, melena, and weight loss are the most common symptoms. The incidence of upper abdominal pain is about 66%, digestive obstruction is 40%, digestive bleeding is 24%, weight loss is 54.4%, and jaundice is 48.1%.

[0004] Duodenal malignant tumors have a high overall malignancy degree and poor prognosis, which is affected by the depth of tumor infiltration, lymph node metastasis, and surgical approach, and the overall 5-year survival rate is about 27%.

[0005] Genome sequencing of primary duodenal cancer revealed mutation rates >5% in eight genes: KRAS, TP53, APC, SMAD4, PIK3CA, ERBB2, BRAF, and FBXW7, with ERBB2 showing a mutation rate >12%. Further studies also found positive results for microsatellite instability (MSI). Currently, there are no suitable targeted therapies for genes such as KRAS, TP53, APC, and SMAD4.

[0006] Ataxia-telangiectasia (AT) is an autosomal recessive genetic disorder. Common clinical symptoms include ataxia, telangiectasia, sensitivity to chemicals and radiation, immunodeficiency, neurodegeneration, and an increased risk of cancer. Symptoms typically appear in early childhood and worsen over time, often leading to death in the 20s due to chronic lung disease or related cancers. AT patients are abnormally sensitive to radiation, and their cells have defects at cell cycle checkpoints, allowing DNA-damaged cells to continue dividing, increasing mutations and genomic instability, and promoting cancer development. Not only do AT patients have a high risk of cancer, but carriers of the AT-causing gene also have an increased risk of certain cancers. The single gene that causes AT is named ATM. The ATM gene encodes a protein with a molecular weight of approximately 350 kDa, belonging to the phosphatidylinositol 3-kinase-like (PI3K) family of serine / threonine protein kinases, which phosphorylate the SQ / TQ sites of more than 700 target substrates. It plays an initiating role in DNA repair signaling, regulates cell cycle checkpoints, and inhibits cell cycle progression by phosphorylating proteins such as p53, Chk1 / 2, NBS1, and RPA. It also plays a key role in telomere maintenance and apoptosis.

[0007] When a DNA double-strand break (DSB) occurs, the exposed DNA ends phosphorylate / acetylate inactive ATM dimers, promoting their activation. Activated ATMs then initiate a series of signaling cascades involving proteins such as BRCA1, CtIP, and γH2AX, promoting DSB repair through homologous recombination.

[0008] ATM exists in both the nucleus and cytoplasm. Nuclear ATM primarily participates in DNA damage repair and cell cycle regulation, while cytoplasmic ATM regulates AKT activity in the insulin response. Furthermore, ATM dysfunction leads to mitochondrial impairment, increased reactive oxygen species production, and T-cell misophagy defects. It also participates in the cGAS / STING pathway, a crucial component of the innate immune system; however, its function in the cytoplasm remains under investigation.

[0009] Patients with hereditary homozygous or pathogenic compound heterozygous ATM mutations will develop acute tumors (AT). Approximately 25% of AT patients will develop cancer, most commonly hematologic malignancies such as acute leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, and T-cell lymphoblastic leukemia. The risk of cancer increases significantly with age. Individuals carrying pathogenic ATM mutation alleles will not develop AT, but they have an increased risk of developing various cancers, including breast cancer, pancreatic cancer, gastric cancer, and prostate cancer. For example, women carrying a single pathogenic ATM mutation have a 5 to 8 times higher risk of developing breast cancer than the general population; it is estimated that 8.8% of breast cancer patients are AT heterozygous. ATM mutations are believed to promote tumorigenesis during cancer development; however, the molecular mechanisms of tumorigenesis in germline carriers of ATM mutations remain to be investigated.

[0010] Genetic testing results reported in the ClinVar database show that genetic alterations to the ATM gene have more than doubled in the past five years. This database contains over 17,000 reports of germline and somatic variations of ATM, including over 4,000 pathogenic variations and nearly 8,000 variations of undetermined significance (VUS). The majority of pathogenic ATM variations are frameshift and nonsense mutations, accounting for 51.7% and 27.9% respectively, leading to protein truncation or complete absence of ATM expression; 97% of VUS are missense mutations.

[0011] The Cancer Genome Atlas (TCGA) pan-cancer atlas study revealed somatic ATM alterations in 6% of cancer tissue samples, with the most frequent alterations observed in uterine cancer (18.6%), bladder cancer (14.11%), and colorectal cancer (12.5%). These mutations are relatively evenly distributed across ATM genes, but the Cancer Hotspots Database lists several recurring mutation sites, such as R250 and R337 at the N-terminus, R1466 in the clamp domain, and R2832, N2875, I2888, L2890, and R3008 in the kinase domain. Understanding the origins and biological impacts of these hotspots is crucial for future research. Summary of the Invention

[0012] The development and progression of duodenal cancer are caused by a variety of factors, among which mutations in key cancer genes are the main cause. Therefore, the purpose of this invention is to provide duodenal cancer cell lines with ATM gene mutations, methods for their isolation and purification, and their uses for cancer and other scientific research.

[0013] On the one hand, the present invention provides a duodenal cancer cell line TH-1, which has the accession number CCTCC No:C2025220.

[0014] In an embodiment of the present invention, the cell line TH-1 has an ATM gene mutation.

[0015] In an embodiment of the present invention, the TH-1 cell line efficiently expresses DHX33.

[0016] In embodiments of the present invention, the TH-1 cell line is highly sensitive to DHX33 helicase inhibitors but insensitive to anticancer drugs. In further embodiments, the DHX33 helicase inhibitor may be KY386, KY38, or other DHX33 inhibitors. DHX33 helicase inhibitors are disclosed in, for example, Chinese patent applications 202011532460.0, 202110724793.1, 202211413921.1, 202211486053.X, 202211486081.1, 202211720769.1, and 202310472343.7.

[0017] In an embodiment of the present invention, cell line TH-1 has a short doubling time and unlimited proliferation capacity.

[0018] In a second aspect, the present invention provides a method for isolating and purifying the TH-1 cell line, the method comprising the following steps:

[0019] S1. Soak duodenal cancer tissue in phosphate buffer and then homogenize the tissue;

[0020] S2. Add trypsin solution and digest at 37°C. Then add complete culture medium containing 10% fetal bovine serum to terminate the digestion.

[0021] S3. Remove the digested single-cell suspension and place it in a carbon dioxide incubator for cell growth using a complete culture medium containing 10% fetal bovine serum and antibiotics; and

[0022] S4. The proliferating, uniformly morphologically homogeneous monoclonal cells were regionally digested with trypsin, and then the cells were expanded and preserved.

[0023] In an embodiment of the present invention, after removing the digested single-cell suspension, the single cells can be seeded into a culture dish and then placed in an incubator for culture.

[0024] In embodiments of the present invention, the complete culture medium may be, for example, DMEM, RPMI-1640, F12K and IMDM medium, preferably F12K medium.

[0025] In embodiments of the present invention, the antibiotic may be penicillin or streptomycin, for example, 1% penicillin and 1% streptomycin.

[0026] In embodiments of the present invention, the concentration of pancreatic enzyme can be 0.05-0.25%, for example 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.015%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, etc.

[0027] In embodiments of the present invention, cell expansion and preservation can be performed, for example, by transferring the cells to a culture dish.

[0028] In a third aspect, the present invention provides a method for screening anticancer drugs, comprising contacting a test compound with TH-1 cells and determining, based on the sensitivity of TH-1 cells to the test compound, whether the test compound can be used to treat duodenal cancer with ATM gene mutation.

[0029] In an embodiment of the present invention, the anticancer drug is a DHX33 helicase inhibitor.

[0030] In a fourth aspect, the present invention provides the use of the duodenal cancer cell line TH-1 in screening anticancer drugs.

[0031] The TH-1 duodenal cancer cell line provided by this invention has an ATM gene mutation and expresses DHX33 efficiently. It can be used as an experimental tool for later cancer or other related scientific research, and can be used to screen anticancer drugs for duodenal cancer with ATM gene mutation. Attached Figure Description

[0032] Figure 1 The expression of DHX33 protein in duodenal cancer pathological tissue was shown.

[0033] Figure 2 The growth morphology of TH-1 cells after isolation is shown.

[0034] Figure 3 The growth curve of TH-1 cells is shown.

[0035] Figure 4 The STR signature map of cell line TH-1 is shown.

[0036] Figure 5 The results show the cell line TH-1's sensitivity to different types of targeted anticancer drugs (cell morphology).

[0037] Figure 6 The study showed the cell line TH-1's sensitivity to different types of targeted anticancer drugs (survival index).

[0038] Figure 7 The results of TH-1 cell genomic DNA sequencing show the ATM gene mutation information.

[0039] The human duodenal cancer cell line TH-1 has been deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, 430072, China, with accession number CCTCC No: C2025220 and deposit date of July 23, 2025. The taxonomic name of this cell line is Homosapiens. Detailed Implementation

[0040] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. However, those skilled in the art should understand that the embodiments described below are only for illustrating the present invention and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0041] 1. Immunohistochemical analysis

[0042] Pathological tissue sections were deparaffinized in xylene and hydrated in a series of solutions with gradually decreasing ethanol concentrations. The antigen was presented in a vaporizer with Tris buffer (pH 9.0). The tissues were then incubated in a methanol solution containing 1% H2O2 to inactivate endogenous peroxidase. After blocking with phosphate-buffered saline (PBS) containing 5% fetal bovine serum for 1 h at room temperature, the tissues were incubated with the primary antibody at 25°C for 2 h. Subsequent immunohistochemical analysis was performed according to the manufacturer's instructions using the DAKO kit (DAKO, K5007). The antibodies used were as follows: primary antibody: anti-DHX33 antibody (B4, purchased from Santa Cruz Biotechnology Co., Ltd.); secondary antibody: ready-to-use MaxVision™ HRP kit (mouse / rabbit), purchased from Fuzhou Maixin Biotechnology Development Co., Ltd.

[0043] 2. Cell Culture

[0044] The isolated TH-1 cells were cultured in F12K medium containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, streptomycin, and penicillin. Human primary HFF cells were cultured in DMEM medium containing 10% FBS, 2 mM L-glutamine, streptomycin, and penicillin, and incubated in a humidified environment containing 5% carbon dioxide.

[0045] 3. Cell growth curve analysis

[0046] Cells were digested with trypsin and resuspended in complete F12K cell culture medium to produce a single-cell suspension. Cells were counted and then seeded in 6-well plates at 50,000 cells per well. Cell counts were calculated daily from the start of plate formation.

[0047] 4. Mouse xenotransplantation model

[0048] All mouse experiments were conducted in accordance with standard guidelines. Balb / cNUDE female mice were purchased from Beijing Victoria Laboratory Animal Co., Ltd. and received standard institutional care. Cells were digested with trypsin and resuspended in PBS to a final concentration of 1 × 10⁻⁶ cells / mL. 8 Cells. 1×10⁻⁶ cells were injected subcutaneously into the lateral flank of 6-week-old nude mice (Balb / c). 7 Each treatment group consisted of 5-10 mice, and the growth of subcutaneous tumors in the mice was analyzed.

[0049] 5. Cellular half-inhibitory concentration (IC50) 50 Value test

[0050] The cancer cell line TH-1 was used at a rate of 1×10 4 100 μL / well of cells were seeded onto a 96-well plate. After complete cell adhesion, various anticancer compounds were added to the cell culture medium at concentrations of 10 nM, 25 nM, 50 nM, 100 nM, 250 nM, 500 nM, 1 μM, 2.5 μM, 5 μM, 10 μM, and 20 μM, and mixed thoroughly using a multichannel pipette. After 48 hours of incubation with the anticancer compounds and cells, the medium was added to the 96-well plate using CCK-8 reagent (Shanghai Yisheng Biotechnology Co., Ltd.) according to standard procedures. After 2 hours of incubation, the plate was read using an ELISA reader (OD). 450nm The experiment was repeated three times, and growth inhibition curves of the compound at different concentrations were plotted. The half-inhibitory concentration (IC50) of the compound in TH-1 cells was calculated. 50 ).

[0051] 6. Sequencing of the ATM gene in the TH-1 cell genome

[0052] DNA extraction was performed according to the standard operating procedure of the Tiangen Cell Genomic DNA Extraction Kit (Tiangen Biotech (Beijing) Co., Ltd., DP304). Then, using genomic DNA from TH-1 cells as a template, PCR amplification was performed using the following primers: ATM gene forward primer: 5'-TACCCCTTGCCAATGGAAGAT-3'; ATM reverse primer: 5'-AGACTGTACCTACCAAAATGCT-3'. The purity of the PCR amplification products was analyzed by gel electrophoresis, and the samples were submitted for gene sequencing.

[0053] 7. STR (short repeat sequence) analysis of TH-1 cells

[0054] DNA extraction was performed according to the standard operating procedure of the Tiangen Cell Genomic DNA Extraction Kit (Tiangen Biotech (Beijing) Co., Ltd., DP304). Then, using TH-1 cell genomic DNA as a template, specific primer PCR amplification was performed using the 21-STR amplification protocol, and the amplified fragments were analyzed and matched. STR loci and the sex gene Amelogenin were detected using a Seqstudio genetic analyzer. The specific STR genotyping protocol is as follows:

[0055] Scheme 1 Scheme 2 Scheme 3 Scheme 4 1 D19S433 AMEL D2S441 TH01 2 D5S818 D3S1358 vWA D12S391 3 D21S11 D13S317 D8S1179 D2S1338 4 D18S51 D7S820 TPOX FGA 5 D6S1043 D16S539 PentaE 6 CSF1PO 7 PentaD

[0056] 8. Statistical Analysis

[0057] Data are expressed as mean ± SD. Statistical significance was determined using Student's t-test, with a p-value < 0.05 indicating a significant difference.

[0058] Example

[0059] The preferred embodiments of the present invention will now be described in detail with reference to specific examples. It should be understood that the following embodiments are provided for illustrative purposes only and are not intended to limit the scope of protection of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from its spirit and intent, and all such modifications and substitutions fall within the scope of protection claimed in the present invention.

[0060] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0061] 1. Analysis of DHX33 protein expression in a duodenal cancer tissue

[0062] Immunohistochemical analysis was performed on paraffin-embedded sections of duodenal cancer tissue (provided by Donghua Hospital). The primary antibody, anti-DHX33 antibody, was purchased from Santa Cruz Biotechnology, clone number B4. The working concentration of the primary antibody was 1:20, diluted with phosphate-buffered saline (PBS) containing 5% fetal bovine serum. The tissue was then washed repeatedly with PBS. The tissue was incubated with ready-to-use secondary antibody at room temperature for 1 hour. After extensive washing with PBS, DAB working solution (Kaiyue Life Sciences, self-developed reagent, Guangdong Shenyang Medical Device Registration No. 20240883) was added. The tissue was then incubated at room temperature for 2 minutes, followed by rapid rinsing with purified water to terminate the colorimetric reaction. Cell nuclei were stained with hematoxylin solution (Kaiyue Life Sciences, self-developed reagent, Guangdong Shenyang Medical Device Registration No. 20240884) for 2 minutes, followed by washing with PBS several times. The stained tissue was dehydrated or appropriately dried, then mounted with resin for microscopic observation and photography.

[0063] The experimental results showed that the cancer cells in the pathological tissue of this cancer exhibited high-efficiency expression of the DHX33 protein.

[0064] 2. Isolation and culture of cell lines

[0065] Biopsy tissue was obtained from duodenal cancer tissue and placed in sterile phosphate buffer using sterile instruments. The tissue was then minced with sterile scissors, resuspended in PBS, and centrifuged to precipitate the tissue. The supernatant was discarded, and 2 mL of 0.25% trypsin solution was added to the precipitated tissue. The tissue was then incubated in a 37°C water bath for digestion, with constant agitation to ensure uniform absorption of the trypsin by the cells. After 5 minutes of trypsin digestion, the digestion solution was allowed to stand, and then 0.1 mL of the cell suspension (free of tissue fragments) was gently pipetted into 10 mL of complete F12K cell culture medium containing 10% fetal bovine serum. The cells were gently agitated to ensure uniform dispersion, and then cultured in a CO2 incubator. The remaining digested cell suspension was added to complete culture medium using the same method. The cell culture medium was refreshed every 3-5 days until individual cancer cells gradually formed a single cell line clone. Single clones were picked for expansion and further culture. The first batch of expanded cells was cryopreserved, and the passage number was designated as P0. If cells from P0 are passaged again later, they are designated as P1, and so on. The process of cell clonal proliferation can be achieved through… Figure 2 exhibit.

[0066] 3. Selection of the optimal cell culture medium

[0067] Four different complete culture media were selected, each containing 10% fetal bovine serum, 1% penicillin, and 1% streptomycin. These four different complete culture media were: DEMM, RPMI-1640, F12K, and IMDM. Equal volumes of 1×10⁻⁶ media were then used. 6 TH-1 cancer cells were resuspended in the four culture media mentioned above and incubated statically in a CO2 incubator. After 3 days of culture, a preliminary assessment of cell proliferation was conducted: F12K medium showed the fastest cell proliferation, the highest cell number, and the highest density, with uniform cell morphology. DMEM medium showed the slowest proliferation and the lowest density, while RPMI-1640 and IMDM media exhibited the least morphological variation. Therefore, F12K medium was ultimately chosen for cell culture.

[0068] 4. Analysis of TH-1 cell proliferation properties

[0069] Equal amounts of TH-1 cells (at a ratio of 1 x 10) 5Cells were seeded into 6-well plates, with the seeding day designated as D0. Cells were then counted on D1, D2, D3, D4, D5, and D6, with each count performed independently three times. The average value was determined, and the overall cancer cell proliferation curve was analyzed. Specific data are as follows: Figure 3 As shown, this TH-1 cell has a relatively rapid proliferation capacity.

[0070] TH-1 cells were passaged every 3 days, with each passage using a 1:5 dilution ratio. This passaged culture continued for 5 months. Analysis showed no significant changes in overall cell morphology. The cell proliferation rate maintained a relatively consistent trend, thus confirming that the TH-1 cell line possesses immortalization and proliferative capabilities.

[0071] 5. Characterization of STR cell lines

[0072] TH-1 cells from generation P0 were cultured and amplified. Genomic DNA was then extracted, and short repeat sequences were amplified using specific STR identification primers (specific primer sequences can be found in: Yi-Hsien Chen, Jon P Connelly, Colin Florian, Xiaoxia Cui, Shondra M Pruett-Miller, 2023, Dis Model Mech 16(10), Short tandem repeat profiling via next-generation sequencing for cell line authentication). The PCR products after amplification were then used for STR gene sequencing to determine the characteristic profile of the STR. The specific experimental protocol is described in section "7. STR (short repeat sequence) analysis of TH-1 cells". The experimental results are as follows: Figure 4 As shown, this result demonstrates that the STR profile of TH-1 is unique and does not highly overlap with existing cancer cell lines.

[0073] 6. Analysis of the inhibitory sensitivity of TH-1 to various targeted anticancer drugs

[0074] Duodenal cancer cell line TH-1 cells were injected at a rate of 1×10 4 Seed 100 μL / well of cells into a 96-well plate. Wait for the cells to adhere completely, then... Figure 6 Different anticancer drugs were added to F12K cell culture medium at concentrations of 20 nM, 100 nM, and 500 nM, with three replicates for each concentration. The mixture was thoroughly mixed using a multichannel pipette. After 48 hours of incubation with the anticancer drugs and cells, the CCK-8 reagent (Shanghai Yisheng Biotechnology Co., Ltd.) was added to the F12K culture medium in 96-well plates according to standard procedures. After 2 hours of incubation, the plates were read using an ELISA reader (OD).450nm The experiment was repeated three times to observe the cell morphology and cell viability of TH-1 cells after the addition of anticancer drugs. Figure 5 The images show representative cell morphology diagrams after treatment with various anticancer drugs. Figure 6 This study presents a comparison of cell viability data after treatment with different concentrations of various anticancer drugs. The results show that the DHX33 inhibitor KY386 exhibits the best TH-1 cell inhibitory activity.

[0075] 7. Identification and analysis of the ATM gene in the TH-1 cell line

[0076] Genomic DNA was extracted from P0 generation TH-1 cells, and then used as a template for gene amplification of a specific region of ATM. The primers used were as described in section "6. Sequencing of the ATM gene in the TH-1 cell genome". After purification, the obtained PCR product was sequenced using the amplification pre-primer, and the sequencing results were as follows: Figure 7 As shown, the experimental results demonstrate that TH-1 cells contain a frameshift mutation in one of the ATM alleles, which prevents ATM from being expressed normally.

[0077] In summary, the duodenal cancer cell line provided in this application has a unique ATM mutation, a relatively unique STR identification profile, and a relatively rapid growth and proliferation capacity, which can be used for cancer biology research such as screening DHX33 helicase inhibitors.

Claims

1. The duodenal cancer cell line TH-1, characterized in that, The cell line has the accession number CCTCC No:C2025220.

2. The cell line TH-1 according to claim 1, characterized in that, The cell line has an ATM gene mutation.

3. The cell line TH-1 according to claim 1, characterized in that, The cell line efficiently expresses DHX33.

4. The cell line TH-1 according to claim 1, characterized in that, The cell line is highly sensitive to DHX33 helicase inhibitors.

5. The cell line TH-1 according to claim 1, characterized in that, The cell line has a short doubling time and unlimited proliferation capacity.

6. A method for isolating and purifying cell line TH-1, characterized in that, Includes the following steps: S1. Soak duodenal cancer tissue in phosphate buffer and then homogenize the tissue; S2. Add trypsin solution and digest at 37°C. Then add complete culture medium containing 10% fetal bovine serum to terminate the digestion. S3. Remove the digested single-cell suspension and place it in a carbon dioxide incubator for cell growth using a complete culture medium containing 10% fetal bovine serum and antibiotics; and S4. The homogeneous monoclonal cells were regionally digested with trypsin, followed by cell expansion and preservation. Preferably, the concentration of the pancreatic enzyme is 0.05-0.25%.

7. The method according to claim 6, characterized in that, The antibiotics mentioned are penicillin and streptomycin.

8. The method according to claim 6, characterized in that, The complete culture medium can be DMEM, RPMI-1640, F12K and IMDM, with F12K being preferred.

9. A method for screening anticancer drugs, comprising contacting a test compound with TH-1 cells and determining whether the test compound can be used to treat duodenal cancer with an ATM gene mutation based on the sensitivity of the TH-1 cells to the test compound.

10. Use of the duodenal cancer cell line TH-1 according to claim 1 in screening anticancer drugs.

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