Primary cell and progeny cell of human lung adenocarcinoma tumor and application of primary cell and progeny cell

Abstract

The invention discloses primary cells and progeny cells of human lung adenocarcinoma tumors and application of the primary cells and the progeny cells. The invention provides a human lung adenocarcinoma primary cell, in particular to a human lung adenocarcinoma primary cell CR110, and the preservation number of the human lung adenocarcinoma primary cell CR110 in the China Center for Type Culture Collection (CCTCC) is CCTCC NO: C202419. The human lung adenocarcinoma primary cell CR110 provided by the invention has in-vitro proliferation ability and stem cell balling ability, has NCG mouse subcutaneous tumor formation ability, and shows different drug toxic reactions to common chemotherapeutic drugs for lung adenocarcinoma. The human lung adenocarcinoma primary cell CR110 provided by the invention can be used for analyzing the pathogenesis of the human lung adenocarcinoma in vitro and in vivo and detecting related characteristics such as drug sensitivity in vitro, and experimental materials closer to clinical tumor biological characteristics are provided for research of the human lung adenocarcinoma.

Description

Technical Field

[0001] The present invention belongs to the field of cell biology, and particularly relates to a primary human lung adenocarcinoma cell, a progeny cell thereof, and their applications. Background Art

[0002] The treatment of cancer is multidisciplinary, and cancer surgery, chemotherapy, and radiotherapy are listed as the three major important treatment methods for cancer. Among them, chemotherapy mainly uses anti-cancer drugs (including molecular targeted drugs) to inhibit the proliferation of cancer cells, thereby shrinking and destroying tumor cells. However, there are significant differences in the biomedical characteristics, drug sensitivities, prognoses, etc. of different tumor patients with the same pathological type. This individual difference also makes the treatment of tumors more complicated and uncertain. Due to the heterogeneity problem of tumors, the use of anti-cancer drugs for tumor patients in clinical practice has a certain degree of blindness and randomness, and the treatment effect is not good, and the side effects are obvious. The mechanism of tumor progression, the efficacy of drugs, and the mechanism of drug resistance generation are still unclear. Traditional drug resistance models are mostly established with cell lines. After multiple passages of the cell line model, both the genotype and phenotype have changed greatly, resulting in a huge difference between in vitro drug sensitivity tests and in vivo drug efficacy, and it is not suitable for extensive clinical application. Therefore, there is an urgent need in clinical practice for an in vitro model that can verify the efficacy individually to predict the response in clinical treatment, so as to provide a more reliable precision medication strategy and prolong the survival period of patients. Summary of the Invention

[0003] The purpose of the present invention is to provide a primary human lung adenocarcinoma cell, a progeny cell thereof, and their applications.

[0004] In the first aspect, the present invention claims to protect a primary human lung adenocarcinoma cell.

[0005] The primary human lung adenocarcinoma cell claimed by the present invention is the primary human lung adenocarcinoma cell CR110, and its deposit number in the China Center for Type Culture Collection is CCTCC NO: C202419.

[0006] The primary human lung adenocarcinoma cell CR110 is a primary human lung adenocarcinoma cell.

[0007] In the second aspect, the present invention claims to protect the progeny cell of the primary human lung adenocarcinoma cell described in the first aspect above.

[0008] The daughter cells have the same STR data as the primary cells (i.e., the primary human lung adenocarcinoma cells CR110 described in the first aspect above), specifically: Amelogen in-X / Y, D3S1358-15 / 17, vWA-14 / 17, D7S820-10 / 11, CSF1PO-9 / 12, PentaE-12 / 14, D8S1179-13 / 13, D21S11-32 / 33.2, D16S539-10 / 11, D2S1338-23 / 24, PentaD-13 / 15, D19S433-13.2 / 13.2, TH01-6 / 9, D13S317-8 / 10, TPOX-9 / 11, D18S51-14 / 19, D6S1043-11 / 20, D1S1656-13 / 17, D5S818-9 / 11, D12S391-18 / 18, FGA-22 / 23, covering 21 loci for general detection.

[0009] In a third aspect, the present invention claims the use of the primary human lung adenocarcinoma cells described in the first aspect above or the daughter cells described in the second aspect above in the construction of a CDX tumor model (CDX model).

[0010] The CDX model, fully known as the Cell line-derived xenograft (CDX) model, is a tumor model constructed by transplanting an in vitro cultured heterologous (human) tumor cell line into an immunodeficient mouse.

[0011] In a fourth aspect, the present invention claims the use of the primary human lung adenocarcinoma cells described in the first aspect above or the daughter cells described in the second aspect above in any of the following:

[0012] (A1) Preparing a product for in vitro study of the pathogenesis of human lung adenocarcinoma;

[0013] (A2) In vitro study of the pathogenesis of human lung adenocarcinoma.

[0014] In a fifth aspect, the present invention claims the use of the primary human lung adenocarcinoma cells described in the first aspect above or the daughter cells described in the second aspect above in any of the following:

[0015] (B1) Preparing a product for in vitro screening of therapeutic drugs for human lung adenocarcinoma;

[0016] (B2) In vitro screening of therapeutic drugs for human lung adenocarcinoma.

[0017] In a sixth aspect, the present invention claims the use of the primary human lung adenocarcinoma cells described in the first aspect above or the daughter cells described in the second aspect above in any of the following:

[0018] (C1) Preparing a product for in vitro study of the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma;

[0019] (C2) In vitro study of the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma.

[0020] In a seventh aspect, the present invention claims protection for a method of constructing a CDX tumor model.

[0021] The method of constructing a CDX tumor model (CDX model) claimed by the present invention may include the following steps: transplanting the primary human lung adenocarcinoma cells described in the first aspect above or the progeny cells described in the second aspect above into immunodeficient mice to obtain the CDX tumor model (CDX model).

[0022] In an eighth aspect, the present invention claims protection for the application of the CDX tumor model constructed by the method described in the seventh aspect above in the study of the pathogenesis of human lung adenocarcinoma.

[0023] In a ninth aspect, the present invention claims protection for the application of the CDX tumor model constructed by the method described in the seventh aspect above in the in vitro screening of therapeutic drugs for human lung adenocarcinoma.

[0024] In a tenth aspect, the present invention claims protection for the application of the CDX tumor model constructed by the method described in the seventh aspect above in the study of the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma.

[0025] Advantages of the present invention: The primary human lung adenocarcinoma cell line CR110 provided by the present invention, identified by STR gene typing, is a primary human lung adenocarcinoma cell line that has never been registered at home and abroad. Karyotype analysis shows that the chromosome number is 58 - 64, and the abnormal ratio is 100%. The cell state is stable, providing experimental materials and models closer to the clinical tumor biological characteristics for the study of human lung adenocarcinoma. The primary cells obtained by the present invention can be directly used as a basic research model for studying the pathogenesis of human lung adenocarcinoma tumors, and can also be used as a screening model for precision drug use plans for human lung adenocarcinoma tumors before clinical trials in vitro.

[0026] Deposit Description

[0027] Classification and naming: Primary human lung adenocarcinoma cell line CR110 Homo sapiens;

[0028] Depositary institution: China Center for Type Culture Collection;

[0029] Abbreviation of the depositary institution: CCTCC;

[0030] Address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province;

[0031] Preservation Date: January 5, 2024;

[0032] Registration Number in the Preservation Center: CCTCC NO: C202419. Description of the Drawings

[0033] Figure 1 It shows the cell morphology of primary human lung adenocarcinoma cell CR110 under an optical microscope. (A) P0 generation cells; (B) P8 generation cells; (C) Spheroid culture of primary human lung adenocarcinoma tumor cell CR110.

[0034] Figure 2 It shows the immunofluorescence identification of primary human lung adenocarcinoma tumor cells. Control represents the negative control, and PBS is used instead of the primary antibody in the immunofluorescence staining steps of primary cells to eliminate false positives of the antibody. The left column is DAPI staining, the middle is antibody staining, and the right column is the staining with DAPI and antibody merged together.

[0035] Figure 3 It shows the karyotype identification diagram of primary human lung adenocarcinoma cell CR110.

[0036] Figure 4 It shows the STR gene typing diagram of the progeny cells of primary human lung adenocarcinoma cell CR110.

[0037] Figure 5 It shows the tumor formation and HE identification of primary human lung adenocarcinoma cell CR110. (A) Tumor formation of primary human lung adenocarcinoma cell CR110; (B) HE identification. Specifically, it is the test result 70 days after inoculating primary human lung adenocarcinoma cell CR110.

[0038] Figure 6 It shows the immunohistochemical identification of tumors in mice with human lung adenocarcinoma. Specifically, it is the test result 70 days after inoculating primary human lung adenocarcinoma cell CR110.

[0039] Figure 7 It shows the drug sensitivity test result diagram of primary human lung adenocarcinoma cell CR110. Detailed Embodiments

[0040] The present invention will be further described in detail below in combination with the detailed embodiments. The provided embodiments are only for clarifying the present invention, rather than limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and do not constitute any limitation to the present invention in any way.

[0041] The experimental methods in the following embodiments, unless otherwise specified, are all conventional methods, and are carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following embodiments, unless otherwise specified, can all be obtained from commercial channels.

[0042] Example 1: Obtaining, Identification and Application of Primary Human Lung Adenocarcinoma Tumor Cells CR110

[0043] I. Experimental Methods

[0044] 1. Isolation, Culture and Subculture of Primary Cells

[0045] A. Isolation and Culture of Primary Cells:

[0046] (1) Quickly rinse the retrieved surgical specimen (lung adenocarcinoma tissue specimen of the patient) with 95%-100% ethanol, then rinse it with PBS at 4°C and transfer it to a new culture dish.

[0047] (2) Cut the tissue into small pieces (<10 mm) with sterilized surgical scissors.

[0048] (3) Take a 15 mL centrifuge tube, add enzyme A, H and R in the Miltenyi human tissue dissociation kit to 4.7 mL of DMEM or RPMI1640 medium, and mix to prepare a digestive enzyme solution.

[0049] (4) Transfer the cut tissue fragments to a 15 mL centrifuge tube with digestive enzymes using a Pasteur pipette, seal it with a sealing film and label the information.

[0050] (5) Transfer it to an incubator at 37°C and culture it on a rotary shaker for 0.5 h - 3 h. Observe the digestion of the tissue fragments every half hour during this period.

[0051] (6) Centrifuge at 500 g, 4°C for 5 min and discard the supernatant.

[0052] (7) Resuspend the precipitate in 10 mL of complete DMEM medium, filter it through a 100 µm filter membrane, and then centrifuge at 300 g, 4°C for 5 min.

[0053] (8) Resuspend the precipitate in primary medium and plate it in a culture plate containing feeder cells, and culture it in an incubator at 37°C and 5% CO2. Refer to 201910212202.5 for completion.

[0054] (9) Observe the cell growth under a microscope every two days and change the medium as needed.

[0055] B. Digestive Subculture:

[0056] (1) When the cells grow to 80%-90% confluence, digest them with 0.05% trypsin / EDTA for 1 min. Observe under the microscope and terminate the digestion when the feeder cells shrink into a round shape, and discard the feeder cells.

[0057] (2) Wash away the residual liquid with PBS, then digest with 0.25% trypsin / EDTA for 1 - 3 min. Stop digestion after all epithelial cells are digested. Resuspend the cells with 5 - 10 mL of complete medium and collect the cells by centrifugation at 500 g for 30 min.

[0058] C. Spheroid culture of primary human lung adenocarcinoma tumor cells:

[0059] Resuspend the digested cells with stem cell medium (Catalog: CTCC-D028 HCM), seed them in an ultra-low attachment well plate, and culture them in an incubator at 37°C with 5% CO2.

[0060] 2. Karyotype identification and STR identification

[0061] Karyotype identification: Select cells in the logarithmic growth phase, add colchicine (final concentration 0.2 μg / mL), shake well and incubate at 37°C for 1 - 2 hours. While waiting, prepare a hypotonic solution of 0.075 M KCl (preparation: 8 mL of sterile water + 44.7 mg of KCl), and preheat it in a 37°C water bath. Digest the incubated cells with trypsin, centrifuge at 1200 rpm for 5 min, collect them in a centrifuge tube, remove the supernatant, resuspend with 8 mL of hypotonic solution, homogenize and place in a 37°C water bath for 40 min. Add 2 mL of freshly prepared fixative (methanol / acetic acid = 2 / 1 to 3 / 1, volume ratio), mix well, let stand at room temperature for 10 min, then centrifuge at 1000 rpm for 10 min, remove the supernatant (leave 500 μL, first resuspend the bottom cells evenly), add 8 mL of fresh fixative, centrifuge at 1200 rpm for 10 min, repeat three times, then drop the cells onto a slide and age in an 80°C oven for 3 hours. Then place the dried slide in freshly prepared trypsin (0.25%) and digest for 1 min (strictly control the time), then place it in Giemsa stain for 5 - 10 minutes (first stain with 1 slide for 5 min, adjust the subsequent staining time according to the color), take it out, rinse off the stain with running water, and observe the results under a microscope.

[0062] STR identification: Send it to an identification company for STR identification.

[0063] 3. Immunofluorescence identification of primary human lung adenocarcinoma tumor cells

[0064] Add 10 5The cells were seeded in a 24-well plate containing cell culture slides. After the cells adhered to the wall the next day, they were rinsed twice with PBS and fixed with 4% paraformaldehyde for 30 min. Primary cells were identified using immunofluorescence. The antibodies used for immunofluorescence were CK7 (Cell Signaling Technology, CST, Cat.#4465T), NapsinA (CST, Cat.#43861S), and TTF-1 (CST, Cat.#12373S).

[0065] 4. Tumorigenicity of primary human lung adenocarcinoma cells and HE identification of CDX tissues

[0066] The digested cells were resuspended in a protective solution (the protective solution was prepared as follows: Mix Matrigel and sterilized normal saline in a volume ratio of 1:2, and then add 1% penicillin-streptomycin double antibody), and transported to the animal house on ice. Select the target animal for local physical hair removal, disinfect after sufficient subcutaneous exposure; in this example, the target animal is severely immunodeficient NCG mice (Guangdong Provincial Center for Medical Experimental Animals, 4 - 6 weeks old, NCG mice); Mix the obtained tumor cells with the protective solution, for every 100 μL of the protective solution, add about 10 7 cell amounts to obtain an injection solution; then, use a 1 mL syringe (needle size 20G) to inject 100 μL of the injection solution containing tumor cells subcutaneously into the target animal.

[0067] HE staining of CDX tissues: (1) Dewax in xylene for 5 - 10 minutes. Replace with fresh xylene solution and dewax for another 5 - 10 minutes. Immerse in absolute ethanol for 5 minutes. Immerse in 90% ethanol for 2 minutes, 80% ethanol for 2 minutes, 70% ethanol for 2 minutes, and immerse in distilled water for 2 minutes. (2) Stain with hematoxylin and eosin (HE) for 5 minutes, adjust according to the state of the slide. Rinse with double-distilled water to remove excess staining solution for about 5 minutes. Wash with distilled water again (control the time). At this time, if direct observation is needed, wash with 70% ethanol twice. If dehydration, clearing and mounting are required, follow the subsequent steps. After washing with 70% ethanol, dehydration, clearing and mounting can still be carried out according to the subsequent steps. (3) Dehydration, clearing, mounting: 70% ethanol for 10 seconds, 80% ethanol for 10 seconds, 90% ethanol for 10 seconds, absolute ethanol for 10 seconds. Clear in xylene for 5 minutes. Replace with fresh xylene and clear for another 5 minutes. Mount with neutral gum or other mounting agents. Observe under the microscope, the cell nuclei are blue, and the cytoplasm is pink or red.

[0068] 5. Immunohistochemical staining of CDX tissues

[0069] (1) Dewaxing / rehydration steps:

[0070] Incubate the sections in xylene twice, 5 minutes each time.

[0071] Incubate the sections in 100% ethanol once, 5 minutes each time.

[0072] Incubate the sections in 95% ethanol once, 5 minutes each time.

[0073] Incubate the sections in 85% ethanol once, 5 minutes each time.

[0074] Incubate the sections in 75% ethanol once, 5 minutes each time.

[0075] Wash twice with PBS, 3 - 5 minutes each time.

[0076] Air dry slightly, and draw small circles around the tissue with an immunohistochemistry pen.

[0077] (2) Antigen retrieval:

[0078] Immerse the sections in 1× citrate retrieval solution and then place them in a microwave oven and heat until boiling; continue to maintain a sub - boiling temperature (95℃ - 98℃) for 10 minutes. Cool the sections on the laboratory bench for 30 minutes.

[0079] (3) Staining:

[0080] Wash the sections with PBS three times, 5 minutes each time.

[0081] Place the sections in 3% hydrogen peroxide aqueous solution and incubate for 10 minutes.

[0082] Wash the sections with PBS twice, 5 minutes each time.

[0083] Wash the sections with wash buffer for 5 minutes continuously.

[0084] Drop 100 μL of blocking solution on each section and block at room temperature for 1 hour.

[0085] Remove the blocking solution, add 100 μL of primary antibody diluted with the recommended antibody diluent. Incubate overnight at 4℃. Remove the antibody solution and wash the sections with wash buffer 3 times, 5 minutes each time.

[0086] Add 1 drop (30 μL) of DAB Chromogen Concentrate to 1 ml of DAB Diluent and mix well before use. Apply 100 μL of DAB on each section and observe closely. Usually, appropriate staining intensity can be obtained in 5 minutes.

[0087] Immerse the sections in PBS. Wash the sections with PBS twice, 5 minutes each time.

[0088] (4) Dehydrate the sections:

[0089] Incubate the sections in 85% ethanol once for 5 minutes each time.

[0090] Incubate the sections in 95% ethanol once for 5 minutes each time.

[0091] Incubate the sections in 100% ethanol once for 5 minutes each time.

[0092] Repeat the incubation in xylene twice for 30 seconds each time.

[0093] (5) Sealing the coverslip: Wipe off the excess liquid around the tissue, and carefully seal the coverslip with neutral balsam. After sealing, fix the periphery of the coverslip with a small amount of clear nail polish.

[0094] (6) Observe, photograph, and save under the microscope.

[0095] 6. Drug sensitivity detection of primary human lung adenocarcinoma cells

[0096] Seed primary human lung adenocarcinoma cells at a density of 10 4 / well in a 96-well plate. After the cells adhere to the wall the next day, add different drugs and treat for 48 h respectively. Detect the OD value of the cells by the CCK8 method and calculate the inhibition rate of the drugs.

[0097] II. Results and analysis

[0098] 1. Culture and passage of primary human lung adenocarcinoma tumor cells

[0099] The isolated tumor cells were co-cultured with feeder cells. Change the culture medium after 3 - 7 days. Obvious cell clones can be observed under the microscope. The tumor cells adhere to the wall and aggregate, arranging tightly in a cobblestone paving stone shape. In the co-culture system, island-like cell clusters are clearly visible and are tightly wrapped and growing by the feeder cells. The feeder cells are spindle-shaped and can be clearly distinguished from epithelial cells. See Figure 1 in (A). After passage of these primary cells, they are consistent with the shape of the primary cells (P0 generation), showing a tightly arranged epithelial-like shape (P8 generation). See Figure 1 in (B). In the stem cell culture system, the morphology of most of these primary cells changes significantly. They change from polygonal when adherent cells to round or oval, and gradually proliferate and aggregate into clusters, forming floating cell spheres with smooth edges. See Figure 1 in (C).

[0100] 2. Immunofluorescence identification of primary human lung adenocarcinoma tumor cells

[0101] The cultured cells were identified by immunofluorescence method, as Figure 2As shown, the immunofluorescence results showed that CK7 was strongly positive, and both NapsinA and TTF-1 were positively expressed. The immunofluorescence antibody expression results were consistent with the clinical pathological results of the patient's tumor, indicating that the cultured primary cells were lung adenocarcinoma tumor cells.

[0102] 3. Karyotype identification

[0103] The karyotype identification results are as Figure 3 shown. The karyotype analysis and identification of primary human lung adenocarcinoma tumor cells showed that the chromosome number was 58 - 64, and the abnormal ratio was 100%.

[0104] 4. STR analysis and identification of human lung adenocarcinoma tumor cells

[0105] The results showed that the STR typing data of the primary human lung adenocarcinoma cells and their progeny cells obtained in the present invention were: Amelogenin-X / Y, D3S1358-15 / 17, vWA-14 / 17, D7S820-10 / 11, CSF1PO-9 / 12, PentaE-12 / 14, D8S1179-13 / 13, D21S11-32 / 33.2, D16S539-10 / 11, D2S1338-23 / 24, PentaD-13 / 15, D19S433-13.2 / 13.2, TH01-6 / 9, D13S317-8 / 10, TPOX-9 / 11, D18S51-14 / 19, D6S1043-11 / 20, D1S1656-13 / 17, D5S818-9 / 11, D12S391-18 / 18, FGA-22 / 23, covering 21 loci for general detection. Figure 4 It is the STR gene typing map of the progeny cells of primary human lung adenocarcinoma cells.

[0106] Comparing with the ATCC data, there was no matching cell line or primary cell, indicating that this primary cell was a new case of primary human lung adenocarcinoma tumor cells, which was named primary human lung adenocarcinoma cell CR110.

[0107] The primary human lung adenocarcinoma cell CR110 was deposited at the China Center for Type Culture Collection on January 5, 2024, and its deposit number was CCTCC NO: C202419.

[0108] 4. Tumor formation and HE identification of primary human lung adenocarcinoma tumor cells in mice

[0109] The primary human lung adenocarcinoma cell CR110 could form tumors in NCG mice, as shown in Figure 5 (A), and the tissue HE identification showed atypical cells. Combining the morphological and immunohistochemical results, it was consistent with lung adenocarcinoma, as shown in Figure 5 (B).

[0110] 5. Identification of tumor immunohistochemistry in human lung adenocarcinoma tumor-bearing mice

[0111] The results of immunohistochemical identification of tumors in CDX mice showed that ( Figure 6 ), the expression of immunohistochemical markers in CDX was consistent with that of the markers in the patient's tumor itself (ck7+, NapsinA+, TTF-1+, p63-), revealing that the primary cells were of human origin and were the only primary cells.

[0112] 6. Drug sensitivity test of primary human lung adenocarcinoma cells

[0113] The main clinical chemotherapy drugs for lung adenocarcinoma include cisplatin, carboplatin, pemetrexed, gemcitabine hydrochloride, paclitaxel, docetaxel, vinorelbine, etc. The drug sensitivity results of primary human lung adenocarcinoma cell CR110 showed that ( Figure 7 ), at different drug concentrations, gemcitabine had the highest inhibition rate on cells, carboplatin had the lowest inhibition rate, and the other chemotherapy drugs also showed different sensitivities, which could be used as an in vitro screening model for drug screening in clinical practice.

[0114] The above has described the present invention in detail. For those skilled in the art, without departing from the purpose and scope of the present invention and without unnecessary experiments, the present invention can be implemented within a relatively wide range under equivalent parameters, concentrations and conditions. Although specific embodiments of the present invention are given, it should be understood that the present invention can be further improved. In short, according to the principle of the present invention, this application intends to cover any changes, uses or improvements to the present invention, including changes made by using conventional techniques known in the art that are outside the scope disclosed in this application.

Claims

1. A primary human lung adenocarcinoma cell, characterized in that: The primary human lung adenocarcinoma cells are primary human lung adenocarcinoma cells CR110, and their preservation number at the China Center for Type Culture Collection is CCTCC NO: C202419.

2. The progeny cells of the primary human lung adenocarcinoma cells described in claim 1.

3. The application of the primary human lung adenocarcinoma cells described in claim 1 or the progeny cells described in claim 2 in constructing a CDX tumor model.

4. The application of the primary human lung adenocarcinoma cells described in claim 1 or the progeny cells described in claim 2 in any of the following: (A1) Preparing a product for studying the pathogenesis of human lung adenocarcinoma; (A2) Studying the pathogenesis of human lung adenocarcinoma.

5. The application of the primary human lung adenocarcinoma cells described in claim 1 or the progeny cells described in claim 2 in any of the following: (B1) Preparing a product for in vitro screening of therapeutic drugs for human lung adenocarcinoma; (B2) In vitro screening of therapeutic drugs for human lung adenocarcinoma.

6. The application of the primary human lung adenocarcinoma cells described in claim 1 or the progeny cells described in claim 2 in any of the following: (C1) Preparing a product for studying the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma; (C2) Studying the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma.

7. A method for constructing a CDX tumor model, comprising the following steps: transplanting the primary human lung adenocarcinoma cells described in claim 1 or the progeny cells described in claim 2 into an immunodeficient mouse to obtain a CDX tumor model.

8. The application of the CDX tumor model constructed by the method described in claim 7 in studying the pathogenesis of human lung adenocarcinoma.

9. The application of the CDX tumor model constructed by the method described in claim 7 in in vitro screening of therapeutic drugs for human lung adenocarcinoma.

10. The application of the CDX tumor model constructed by the method described in claim 7 in studying the action targets and / or pharmacological mechanisms of therapeutic drugs for human lung adenocarcinoma.

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