Malignant phenotype large cell lung cancer cell strain and application thereof
By providing six malignant phenotypes of large-cell lung cancer cell lines, the problem of lack of stable and easy passage of cell models in the prior art is solved, and the effect of stably culturing and studying the proliferation and migration ability of lung cancer cells in the laboratory is achieved.
Patent Information
- Application Number
- CN202510128641.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2022-08-01
- Publication Date
- 2025-05-09
AI Technical Summary
The prior art is difficult to effectively study and treat malignant lung cancer, especially in the study of lung cancer cell proliferation and migration invasion ability, lacking a stable and prone cell model.
Six malignant phenotypes of large cell lung cancer cell lines are provided, including 801D-A10, 801D-F4, 801D-H10, 801D-E6, 801D-F10 and 801D-F11, ensuring the stability and rapid growth of these cell lines during passage through specific screening and culture methods.
These cell lines are able to pass through stably, have different proliferation and migration invasion abilities, provide good cell models for studying the proliferation and metastasis mechanisms of lung cancer, and are easy to cultivate and promote in traditional cell biology laboratories.
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Figure CN119955728A_ABST
Abstract
Description
[0001] This application is a divisional application of the patent application with application number 202210918627.X, application date August 1, 2022, and invention name “Malignant phenotype large cell lung cancer cell line and its application”. Technical Field
[0002] The invention relates to the field of biotechnology, and in particular to a malignant phenotype large cell lung cancer cell line and an application thereof. Background Art
[0003] Lung cancer is one of the malignant tumors that seriously threatens human health. Its morbidity and mortality rates have increased significantly in the past 50 years. The morbidity and mortality rates of men rank first among all malignant tumors, while the morbidity and mortality rates of women rank second. The cause of lung cancer is still not fully understood. Moreover, more than 50% of lung cancer patients are in the advanced stage when diagnosed, and the prognosis of patients with advanced lung cancer is extremely poor, with a 5-year survival rate of only 5%. Although targeted therapy and immunotherapy have significantly prolonged the survival time of patients in recent years, the frequent occurrence of targeted therapy resistance and adverse reactions of immunotherapy have made the treatment of lung cancer face difficulties. Therefore, exploring the molecular mechanisms and molecular therapeutic targets of the occurrence and development of lung cancer remains an urgent problem to be solved.
[0004] Cells are the basic units of structure and function of organisms and the basis of individual development and system development of organisms. Tumor cells have the characteristics of unlimited proliferation, loss of contact inhibition, and weakened cell-to-cell adhesion. In vitro culture and establishment of human lung cancer cell lines or cell strains have very important theoretical and clinical significance for studying the molecular mechanisms and biological characteristics of lung cancer carcinogenesis, invasion and metastasis, and multidrug resistance, as well as the development of new anti-lung cancer drugs. Summary of the invention
[0005] The object of the present invention is to provide a malignant phenotype large cell lung cancer cell line which is easy to culture, stable in passage, easy to popularize and can be used in traditional cell biology laboratories.
[0006] To achieve the above object, the present invention adopts the following technical solution:
[0007] In a first aspect, the present invention provides a large cell lung cancer cell line with a malignant phenotype, wherein the cell line comprises:
[0008] 801D-A10 with the deposit number of CGMCC NO:45140;
[0009] 801D-F4 with the deposit number of CGMCC NO:45141;
[0010] 801D-H10 with the deposit number of CGMCC NO:45139;
[0011] 801D-E6 with the deposit number of CGMCC NO:45138;
[0012] 801D-F10 with the deposit number of CGMCC NO:45142
[0013] and any one of 801D-F11 with a preservation number of CGMCC NO:45143.
[0014] In a specific embodiment, among the cell lines, 801D-A10, 801D-F4 and 801D-H10 cell lines have strong proliferation, migration and invasion abilities, while 801D-E6, 801D-F10 and 801D-F11 cell lines have weak proliferation, migration and invasion abilities.
[0015] In a second aspect, the present invention provides a method for obtaining a large cell lung cancer cell line with a malignant phenotype, the method comprising:
[0016] Human giant cell lung cancer cells 801D were seeded on the upper layer of the Transwell chamber. After the cancer cells were cultured, they were separated by the polycarbonate membrane into three types: cancer cells 801D attached to the polycarbonate membrane, cancer cells 801D below that passed through the polycarbonate membrane but did not grow attached to the membrane and fell to the bottom of the well plate, and cancer cells 801D above that did not pass through the polycarbonate membrane.
[0017] The three cell types were cultured separately, digested, resuspended, and then cultured and expanded as single cells, and monoclonal clones 801D-A10, 801D-F4, 801D-H10, 801D-E6, 801D-F10 and 801D-F11 were screened.
[0018] In a specific embodiment, in the cell line:
[0019] 801D-A10 and 801D-F4 are single clones of 801D that grew adherently through the polycarbonate membrane;
[0020] 801D-E6 and 801D-H10 are monoclonal clones of 801D that did not pass through the polycarbonate membrane;
[0021] 801D-F10 and 801D-F11 are single clones of 801D that passed through the polycarbonate membrane and landed on the bottom of the well plate.
[0022] In a third aspect, the present invention provides the use of a large cell lung cancer cell line with a malignant phenotype in lung cancer research.
[0023] In a fourth aspect, the present invention provides the use of a large cell lung cancer cell line with a malignant phenotype in the study of the proliferation, migration and invasion capabilities of lung cancer cells.
[0024] In a fifth aspect, the present invention provides a cell model for studying lung cancer cell proliferation and metastasis, wherein the cell model comprises any one of 801D-A10, 801D-F4, 801D-H10, 801D-E6, 801D-F10 and 801D-F11 cell lines.
[0025] In a specific embodiment, among the cell lines, 801D-A10, 801D-F4 and 801D-H10 cell lines have strong proliferation, migration and invasion abilities.
[0026] The proliferation, migration and invasion abilities of 801D-E6, 801D-F10 and 801D-F11 cell lines were weak.
[0027] The six human large cell lung cancer cell lines established by the present invention are 801D-A10, 801D-F4, 801D-H10, 801D-E6, 801D-F10 and 801D-F11 cell lines. The six cells can be stably propagated and grow rapidly. In vivo and in vitro experiments confirm that the proliferation, migration and invasion abilities of 801D-A10, 801D-F4 and 801D-H10 cell lines are strong, while in vivo and in vitro experiments confirm that the proliferation, migration and invasion abilities of 801D-E6, 801D-F10 and 801D-F11 cell lines are weak.
[0028] The six human large cell lung cancer cell lines provided by the invention provide good cell models for research related to lung cancer proliferation and metastasis.
[0029] Compared with the prior art, the advantages of the malignant phenotype large cell lung cancer cell line provided by the present invention are:
[0030] (1) Easy to cultivate and promote.
[0031] (2) Ability to be stably propagated.
[0032] Cell deposit information:
[0033] The malignant phenotype large cell lung cancer cell strain provided by the present invention is obtained by screening by the inventor of the present invention and is deposited in the China General Microbiological Culture Collection Administration Center. The deposited names of the malignant phenotype large cell lung cancer cell strains are: human lung cancer cell strain 801D-E6, human lung cancer cell strain 801D-A10, human lung cancer cell strain 801D-F4, human lung cancer cell strain 801D-H10, human lung cancer cell strain 801D-F10, human lung cancer cell strain 801D-F11, and the deposit numbers are CGMCC NO.45138, CGMCC NO.45140, CGMCC NO.45141, CGMCC NO.45139, CGMCC NO.45142, and CGMCC NO.45143. The deposit date is March 30, 2022, the deposit unit is the General Microbiological Center of the China Microbiological Culture Collection Administration Committee, and the deposit unit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The malignant phenotype large cell lung cancer cell line was identified as alive by the General Microbiology Center of China Culture Collection Administration on March 30, 2022. BRIEF DESCRIPTION OF THE DRAWINGS
[0034] Figure 1 The present invention provides 6 types of human large cell lung cancer cell lines with cell morphology.
[0035] Figure 2 The figures are the growth curves of 6 types of human large cell lung cancer cells provided by the present invention.
[0036] Figure 3 The colony formation conditions of 6 human large cell lung cancer cell lines provided by the present invention are shown.
[0037] Figure 4 These are general pictures of nude mouse transplanted tumors of 6 types of human large cell lung cancer cell lines provided by the present invention.
[0038] Figure 5 The six human large cell lung cancer cell lines provided by the present invention and the tumor weights of nude mice transplanted tumors of different cell lines.
[0039] Figure 6 This is the comparison result of the metastasis ability of 801D-A10 cells provided by the present invention and other cell lines.
[0040] Figure 7 This is the comparison result of the metastasis ability of 801D-F4 cells provided by the present invention and other cell lines.
[0041] Figure 8 The results are compared between the metastasis ability of the 801D-H10 cells provided by the present invention and other cell lines.
[0042] Fig. 9This is the comparison result of the metastasis ability of 801D-E6 cells provided by the present invention and other cell lines.
[0043] Fig.10 This is the comparison result of the metastasis ability of 801D-F10 cells provided by the present invention and other cell lines.
[0044] Fig.11 This is a comparison result of the metastasis ability of the 801D-F11 cells provided by the present invention and other cell lines.
[0045] Figures 6 to 11 In the figure, Figures 1, 2, and 3 on the left are pictures of lung metastasis of different cell lines; Figures 1, 2, and 3 on the right are pictures of metastatic lesions in lung tissue sections of different cell lines; the arrows in Figures 1, 2, and 3 on the left point to mice with cachexia, and the arrows in Figures 1, 2, and 3 on the right point to metastatic lesions. DETAILED DESCRIPTION
[0046] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is further described in detail in the following examples. The following examples are only used to illustrate the invention but are not used to limit the scope of the present invention.
[0047] illustrate:
[0048] The cell culture conditions were: 37°C, 5% CO2 static culture.
[0049] The basal medium used in the examples is: serum-free RPMI-1640 medium, fetal bovine serum is added to a concentration of 10%, penicillin is added to a concentration of 1%, and streptomycin is added to a concentration of 1%. 0.2% BSARPMI-1640 medium is: serum-free RPMI-1640 medium is added to a concentration of 0.2% bovine serum albumin (BSA), penicillin is added to a concentration of 1%, and streptomycin is added to a concentration of 1%.
[0050] Example 1
[0051] Cell culture and isolation of different cell lines across Matrigel-coated Polycarbonate membranes
[0052] 1. Cell culture, passaging and inoculation
[0053] 1. 801D cells were inoculated in basal medium.
[0054] 2. Subculture the cells that have grown to 90% confluence, add trypsin digestion solution, and digest at 37°C for about 5 minutes until the cells are completely digested into single cells.
[0055] 3. Add complete culture medium and mix thoroughly by pipetting. After mixing, count the cells using a Muse cell counter.
[0056] 4. Inoculate 801D cells in a 6-well plate, with 500,000 cells per well and each cell type inoculated in 3 wells.
[0057] 5. Incubate the culture medium at 37°C and 5% CO2 overnight.
[0058] 2. Separation of different cell lines through Matrigel-coated Polycarbonate membrane
[0059] 1. Inoculate cells in 6-well plates and observe on the second day. The cell confluence is about 30-40%.
[0060] 2. Aspirate the supernatant in each well and replace with 0.2% BSARPMI-1640 culture medium and continue to culture at 37°C and 5% CO2 for 24 hours.
[0061] 3. Coat the upper chamber surface of the bottom membrane of a Transwell chamber with a pore size of 8.0 μm and a diameter of 6.5 mm with Matrigel and 0.2% BSARPMI-1640 culture medium at a ratio of 1:5, and allow to solidify at 37°C overnight.
[0062] 4. Add 100 μl of 0.2% BSAR PMI-1640 medium to each well of the upper chamber of the Transwell chamber for hydration for 30 minutes, and add 600 μl of basal medium to the lower chamber. Subculture the cells, add trypsin digestion solution, digest at 37°C for about 5 minutes until the cells are completely digested into single cells, add complete medium and mix by pipetting, and count them using a Muse cell counter after mixing.
[0063] 5. Resuspend 801D cells with 0.2% BSARPMI-1640 to a cell concentration of 2 million / ml. Aspirate 100 μl of 0.2% BSARPMI-1640 medium in the Transwell chamber and add 100 μl of the cell suspension to each well. Incubate at 37°C and 5% CO2 for 48 to 72 hours.
[0064] 6. At this time, the cells that pass through the Matrigel-coated Polycarbonate membrane will be divided into two parts, one is attached to the Polycarbonate membrane and grows on the wall, and the other is the cells that pass through the Polycarbonate membrane without growing on the wall and fall on the bottom of the 24-well plate. Digest the cells that grow on the Polycarbonate membrane with trypsin and continue to culture; continue to culture the cells that fall on the bottom of the 24-well plate. In addition, aspirate the cells in the upper chamber of the Transwell chamber that have not passed through the Polycarbonate membrane and continue to culture. In this way, three types of cells are obtained: cells in the upper chamber, cells attached to the membrane, and cells that fall on the bottom of the 24-well plate, which are named: 801D upper, 801D membrane, and 801D lower.
[0065] 3. Screening of three types of 801D cell monoclonal cell lines
[0066] 1. 801D upper, 801D membrane and 801D lower cells were cultured in basal medium until the confluence was 70% to 80%.
[0067] 2. Add trypsin digestion solution and digest at 37℃ for about 5 minutes until the cells are completely digested into single cells. Add basal culture medium, mix well, centrifuge and discard the supernatant. Resuspend with basal culture medium and count with Muse cell counter after resuspension.
[0068] 3. Dilute the culture medium according to the counting results, and the final dilution is 10 cells / ml.
[0069] 4. After mixing, inoculate into a 96-well plate with 100 μl of cell suspension in each well.
[0070] 5. Incubate at 37°C and 5% CO2 and observe after one week.
[0071] 6. Select the wells with only one cell in each well and observe them continuously until they are ready for passage and expansion.
[0072] 7. Six clones were obtained from the single clones screened from 801D upper, 801D membrane and 801D lower, which are:
[0073] 801D: 801D-E6 and 801D-H10;
[0074] 801D film: 801D-A10 and 801D-F4;
[0075] 801D: 801D-F10 and 801D-F11.
[0076] Example 2
[0077] Comparison of cell proliferation ability among different cell lines
[0078] 1. Cell growth curve experiment
[0079] 1. All cells were cultured in basal medium to 70%-80% confluence.
[0080] 2. Add trypsin digestion solution and digest at 37℃ for about 5 minutes until the cells are completely digested into single cells. Add basal culture medium to neutralize, mix well, centrifuge and discard the supernatant. Resuspend with basal culture medium and count with Muse cell counter after resuspension.
[0081] 3. Dilute according to the counting results, and the final dilution is 30,000 cells / ml.
[0082] 4. After mixing, inoculate into 96-well plates, with 5 plates for each group of cells, 5 wells for each cell type at each time point, 3000 cells / 100 μl of cell suspension in each well, and incubate at 37°C, 5% CO2.
[0083] 5. After 0h, 24h, 48h, 72h and 96h, the cell viability was detected by CCK8: at 0h, 10μl CCK8 was directly added to each well, and the absorbance at 450nm was measured by a microplate reader after incubation at 37℃ for 2h; in the other four time periods, the working solution was prepared by adding 10μl CCK8 to 100μl basal culture medium, and the absorbance at 450nm was measured by a microplate reader after incubation at 37℃ for 2h.
[0084] 6. Draw a cell growth curve based on the absorbance values at 5 time points. The results are as follows: Figure 2 shown.
[0085] Compared with 801D-E6 and 801D-F11 cells, the growth rates of 801D-A10 cells and 801D-F4 cells were similar, and the growth rates were significantly increased, with statistical differences (p<0.05).
[0086] Compared with 801D-E6 and 801D-F11 cells, the growth rates of 801D-F4 cells and 801D-H10 cells were similar, and the growth rates were significantly increased, with statistical differences (p<0.05).
[0087] Compared with 801D-F4 and 801D-H10 cells, the growth rates of 801D-E6 cells were similar to those of 801D-F10 cells, but the growth rates of both cells were significantly slower, with statistical differences (p<0.05).
[0088] Compared with 801D-E6 and 801D-F11 cells, the growth rates of 801D-H10 cells and 801D-A10 cells were similar, and the growth rates were significantly increased, with statistical differences (p<0.05).
[0089] Compared with 801D-F4 and 801D-H10 cells, the growth rates of 801D-F10 cells and 801D-F11 cells were similar, and the growth rates were significantly slower, with statistical differences (p<0.05).
[0090] Compared with 801D-F4 and 801D-A10 cells, the growth rates of 801D-F11 cells were similar to those of 801D-F10 cells, but the growth rates of both cells were significantly slower, with statistical differences (p<0.05).
[0091] 2. Colony Formation Assay
[0092] 1. All cells were cultured in basal medium to 70% to 80% confluence.
[0093] 2. Add trypsin digestion solution and digest at 37℃ for about 5 minutes until the cells are completely digested into single cells. Add basal culture medium, mix well, centrifuge and discard the supernatant. Resuspend with basal culture medium and count with Muse cell counter after resuspension.
[0094] 3. Dilute according to the counting results, and the final dilution is 3000 cells / ml.
[0095] 4. After mixing, inoculate into 6-well plates, with three wells for each cell type, containing 300 cells / 2 ml / well of basal culture medium.
[0096] 5. Incubate at 37°C and 5% CO2 and observe after 7 to 9 days.
[0097] 6. Discard the cell culture supernatant, wash twice with PBS, add 2 ml of 2% gentian violet staining solution, and stain for 30 minutes.
[0098] 7. Rinse slowly with tap water to remove excess gentian violet dye and air dry. Figure 3 shown.
[0099] 8. Count colonies with more than 50 cells under a microscope. The results show:
[0100] The colony numbers of 801D-A10 and 801D-F4 cells were significantly higher than those of 801D-E6 and 801D-F11 cells (P<0.05).
[0101] The colony numbers of 801D-F4 and 801D-H10 cells were significantly higher than those of 801D-E6 and 801D-F11 cells (P<0.05).
[0102] The colony numbers of 801D-F4 and 801D-H10 cells were significantly higher than those of 801D-E6 and 801D-F10 cells (P<0.05).
[0103] The colony numbers of 801D-H10 and 801D-A10 cells were significantly higher than those of 801D-E6 and 801D-F11 cells (P<0.05).
[0104] The colony numbers of 801D-F4 and 801D-H10 cells were significantly higher than those of 801D-F10 and 801D-F11 cells (P<0.05).
[0105] The colony numbers of 801D-F4 and 801D-A10 cells were significantly higher than those of 801D-F11 and 801D-F10 cells (P<0.05).
[0106] 3. Subcutaneous tumor formation experiment in BALB / c nude mice
[0107] 1. Culture different cells in basal medium to 70% to 80% confluence.
[0108] 2. Add trypsin digestion solution and digest at 37℃ for about 5 minutes until the cells are completely digested into single cells. Add basal culture medium, mix well, centrifuge and discard the supernatant. Resuspend with basal culture medium and count with Muse cell counter after resuspension.
[0109] 3. Dilute according to the counting results, and the final dilution is 1.0×107 / ml.
[0110] 4. Prepare a corresponding number of BALB / c female nude mice, aged 4-6 weeks.
[0111] 5. After the cell suspension is mixed, it is inoculated into the armpits of nude mice. Each site is injected with 2 million / 200ul cells of the cell suspension, and 5 to 6 mice are inoculated with each cell type.
[0112] 6. Observe the tumor formation in the nude mouse armpits every 3 or 4 days, measure its long and short diameters, and calculate its volume.
[0113] 7. When the volume of the subcutaneous tumor is close to 1000mm 3 When the mice were killed by cervical dislocation, the tumors were dissected out, weighed and photographed.
[0114] 8. Analyze the data based on tumor volume and weight.
[0115] Figure 4 and Figure 5 As shown, compared with 801D-E6 and 801D-F11 cells, the growth rate of 801D-A10 and 801D-F4 cell transplanted tumors was significantly increased (P<0.05), and the subcutaneous tumor formation ability was significantly enhanced (P<0.05).
[0116] Compared with 801D-E6 and 801D-F11 cells, the growth rate of 801D-F4 and 801D-H10 cell transplanted tumors was significantly increased (P<0.05), and the subcutaneous tumor formation ability was significantly enhanced (P<0.05).
[0117] Compared with 801D-H10 and 801D-F4 cells, the growth rate of 801D-F10 and 801D-E6 cell transplanted tumors was significantly slowed down (P<0.05), and the subcutaneous tumor formation ability was significantly reduced (P<0.05).
[0118] Compared with 801D-E6 and 801D-F11 cells, the growth rate of 801D-H10 and 801D-A10 cell transplanted tumors was significantly increased (P<0.05), and the subcutaneous tumor formation ability was significantly enhanced (P<0.05).
[0119] Compared with 801D-H10 and 801D-F4 cells, the growth rate of transplanted tumors of 801D-F10 and 801D-F11 cells was significantly slowed down (P<0.05), and the subcutaneous tumor formation ability was significantly reduced (P<0.05).
[0120] Compared with 801D-A10 and 801D-F4 cells, the growth rate of 801D-F11 and 801D-F10 cell transplanted tumors was significantly slowed down (P<0.05), and the subcutaneous tumor formation ability was significantly reduced (P<0.05).
[0121] Example 3
[0122] Evaluation of the metastatic ability of six types of human large cell lung cancer cells
[0123] 1. Culture different cells in basal medium to 70% to 80% confluence.
[0124] 2. Add trypsin digestion solution and digest at 37℃ for about 5 minutes until the cells are completely digested into single cells. Add basal culture medium, mix well, centrifuge and discard the supernatant. Resuspend with basal culture medium and count with Muse cell counter after resuspension.
[0125] 3. Wash the cells three times with PBS, dilute according to the counting results, and finally dilute with PBS to 1.0×107 / ml.
[0126] 4. Prepare a corresponding number of NOD / SCID female mice, aged 4 to 6 weeks.
[0127] 5. After the cell suspension is mixed, lung cancer cells are inoculated through the tail vein. Each mouse is injected with 0.2 ml (2 million) of cell suspension, and 5 mice are inoculated in each group.
[0128] 6. Every 7 days, luciferase was injected intraperitoneally at a dose of 150 mg / kg. Fifteen minutes later, lung metastasis of mice was observed using a small animal in vivo imaging system.
[0129] 7. At 1.5 to 2 months of age, the mice were killed by cervical dislocation, and the lung tissues were dissected, photographed, fixed with formalin, and stained with HE to observe lung metastasis.
[0130] like Figure 6 As shown, in vivo imaging of small animals showed that 4 mice in the 801D-A10 and 801D-F4 groups had metastatic foci, while no metastatic foci were found in the 801D-E6 and 801D-F11 groups. On the 28th day, one mouse in the 801D-F4 group developed cachexia and a tumor appeared in the left thigh. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-A10 and 801D-F4 groups, while no metastatic foci were found in the 801D-E6 and 801D-F11 groups. This shows that compared with 801D-E6 and 801D-F11 cells, the metastatic ability of 801D-A10 and 801D-F4 cells is enhanced.
[0131] like Figure 7 As shown, in vivo imaging of small animals showed that 4 mice in the 801D-F4 and 801D-H10 groups had metastatic foci, while no metastatic foci were found in 801D-E6 and 801D-F11. On the 28th day, one mouse in the 801D-F4 group developed cachexia and a tumor appeared in the left thigh. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-F4 and 801D-H10 groups, while no metastatic foci were found in 801D-E6 and 801D-F11. This shows that compared with 801D-E6 and 801D-F11 cells, the metastatic ability of 801D-F4 and 801D-F11 cells is weakened.
[0132] like Figure 8 As shown, in vivo imaging of small animals showed that 4 mice in the 801D-F4 and 801D-H10 groups had metastatic foci. In the 801D-E6 and 801D-F10 groups, 1 mouse in the 801D-F10 group had metastatic foci, and no other mice had metastatic foci. On the 28th day, one mouse in the 801D-F4 group had cachexia and a tumor in the left thigh. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-F4 and 801D-H10 groups. In the 801D-E6 and 801D-F10 groups, 1 mouse in the 801D-F10 group had metastatic foci, and no other mice had metastatic foci. This shows that compared with 801D-H10 and 801D-F4 cells, the metastatic ability of 801D-E6 and 801D-F10 cells was weakened.
[0133] like Fig. 9As shown, in vivo imaging of small animals showed that 4 mice in the 801D-H10 and 801D-A10 groups had metastatic foci, while no metastatic foci were found in 801D-E6 and 801D-F11. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-H10 and 801D-A10 groups. This shows that compared with 801D-E6 and 801D-F11 cells, the metastatic ability of 801D-H10 and 801D-A10 cells is enhanced.
[0134] like Fig.10 As shown, in vivo imaging of small animals showed that 4 mice in the 801D-F4 and 801D-H10 groups had metastatic foci. In the 801D-F10 and 801D-F11 groups, 1 mouse in the 801D-F10 group had metastatic foci, and no other mice had metastatic foci. On the 28th day, one mouse in the 801D-F4 group had cachexia and a tumor in the left thigh. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-F4 and 801D-H10 groups. In the 801D-F10 and 801D-F11 groups, 1 mouse in the 801D-F10 group had metastatic foci, and no other mice had metastatic foci. This shows that compared with 801D-H10 and 801D-F4 cells, the metastatic ability of 801D-F10 and 801D-F11 cells was weakened.
[0135] like Fig.11 As shown, in vivo imaging of small animals showed that 4 mice in the 801D-F4 and 801D-A10 groups had metastatic foci, while no metastatic foci were found in the 801D-E6 and 801D-F11 groups. On the 28th day, one mouse in the 801D-F4 group developed cachexia and a tumor appeared in the left thigh. HE staining showed that multiple metastatic foci appeared in the lungs of the 801D-F4 and 801D-A10 groups, while no metastatic foci were found in the 801D-F11 and 801D-F10 groups. This shows that compared with 801D-H10 and 801D-F4 cells, the metastatic ability of 801D-F11 and 801D-F10 cells was weakened.
[0136] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various changes can be made to the technical scheme of the present invention. These simple variations all belong to the protection scope of the present invention.
[0137] It should also be noted that the various specific technical features and steps described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.
[0138] In addition, various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.
Claims
1. A large cell lung cancer cell line with a malignant phenotype, comprising one or more of the following deposited cell lines: The cell line with the accession number of CGMCC NO:45141; The cell line with the accession number of CGMCC NO:45139; The cell line with the accession number of CGMCC NO:45138; A cell line with the deposit number of CGMCC NO:45142; and The cell line with the accession number of CGMCC NO:45143.
2. The large cell lung cancer cell line with a malignant phenotype according to claim 1, comprising a cell line with a preservation number of CGMCCNO:45141.
3. The large cell lung cancer cell line with a malignant phenotype according to claim 1, which is a cell line with a preservation number of CGMCCNO:45141.
4. Use of the large cell lung cancer cell line with a malignant phenotype according to any one of claims 1 to 3 in preparing a cell model for studying lung cancer cell proliferation and metastasis.
5. Use of the large cell lung cancer cell line with a malignant phenotype according to any one of claims 1 to 3 in lung cancer research.
6. Use of the cell line according to any one of claims 1 to 3 in the study of proliferation, migration and invasion ability of lung cancer cells.
7. Use of the large cell lung cancer cell line with a malignant phenotype according to any one of claims 1 to 3 in preparing a small animal model for lung cancer research.
8. The use according to any one of claims 4 to 7, wherein: The malignant phenotype large cell lung cancer cell line is used for subcutaneous tumor formation in BALB / c nude mice.
9. The use according to any one of claims 4 to 7, wherein: The malignant phenotype large cell lung cancer cell line is used for inoculating small animals via the tail vein.
10. A cell model used in an in vitro experiment for studying lung cancer cell proliferation and metastasis, wherein: The cells of the cell model are selected from one or more of the cell lines described in claim 1.