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CN-122012400-A - Lung adenocarcinoma cell line, lung adenocarcinoma animal model and application

CN122012400ACN 122012400 ACN122012400 ACN 122012400ACN-122012400-A

Abstract

The application relates to the technical field of biology, in particular to a lung adenocarcinoma cell line, a lung adenocarcinoma animal model and application. The provided lung adenocarcinoma cell line carries at least one of a group I gene mutation or a group II gene mutation with clinical significance. The in vitro research (such as drug sensitivity test and signal path analysis) of the cell strain can more directly and truly reflect the biological behavior and drug response of lung adenocarcinoma with specific gene mutation background, and provides a highly clinically relevant and standardized in vitro research tool for targeted drug development, combined drug strategy exploration and drug resistance mechanism research.

Inventors

  • YE QIUHUA
  • ZHANG XI
  • GUO NING
  • Peter Edward Robbie

Assignees

  • 清华大学深圳国际研究生院
  • 深圳湾实验室

Dates

Publication Date
20260512
Application Date
20260127
Priority Date
20251021

Claims (15)

  1. 1. A lung adenocarcinoma cell line, characterized in that the lung adenocarcinoma cell line carries a gene mutation comprising at least one of a clinically significant class I gene mutation or a class II gene mutation.
  2. 2. The lung adenocarcinoma cell line of claim 1, wherein the group I gene mutations comprise deletion mutations of EGFR gene and/or, The class II gene mutation comprises at least one of a nonsense mutation of the TP53 gene or a nonsense mutation carrying the PTEN gene.
  3. 3. The lung adenocarcinoma cell line of claim 2, wherein the deletion mutations of the EGFR gene include exon 19 deletion mutations.
  4. 4. A lung adenocarcinoma cell line according to claim 3, characterized in that the deletion mutation of exon 19 results in the deletion of glutamic acid at position 746 to alanine at position 750 in the protein encoded thereby.
  5. 5. The lung adenocarcinoma cell line according to claim 2, characterized in that the nonsense mutation of the TP53 gene results in the premature formation of a stop codon at amino acid 91 of the encoded protein.
  6. 6. The lung adenocarcinoma cell line according to claim 2, characterized in that the nonsense mutation of the PTEN gene causes the protein encoded thereby to form a stop codon in advance at amino acid 319.
  7. 7. The lung adenocarcinoma cell line of any of claims 1-6, characterized in that the cell line comprises at least 1 personality site; optionally, the genotyping of the 1 personality locus is Amelogenin: X, X; Optionally, the cell line comprises at least 8 core STR sites; Optionally, the 8 core STR sites comprise vWA:17,17, D7S820:10,11, CSF1PO:11,11, D16S539:11,11, TH01:9,9, D13S317:8,9, TPOX:8,8, D5S818:9,10; optionally, the cell line comprises at least one of the following STR sites ：Amelogenin：X,X;vWA：17,17;D7S820：10,11;CSF1PO：11,11; D16S539：11,11;TH01：9,9;D13S317：8,9;TPOX：8,8; D5S818：9,10; Optionally, the cell line includes all of the following STR sites ：Amelogenin：X,X;vWA：17,17;D7S820：10,11;CSF1PO：11,11; D16S539：11,11;TH01：9,9;D13S317：8,9;TPOX：8,8; D5S818：9,10.
  8. 8. The lung adenocarcinoma cell line of claim 7, wherein the cell line further comprises at least one of the following STR sites ：D3S1358：16,16;PentaE：5,18,19;D8S1179：10,14,15;D21S11：31.2,32.3;D2S1338：21,22,23;PentaD：10,15;D19S433：13,13;D18S51：14,14;D6S1043：10,10;D1S1656：14,16;D12S391：20,21;FGA：23,24; Optionally, the cell line further comprises all of the following STR sites ：D3S1358：16,16;PentaE：5,18,19;D8S1179：10,14,15;D21S11：31.2,32.3;D2S1338：21,22,23;PentaD：10,15;D19S433：13,13;D18S51：14,14;D6S1043：10,10;D1S1656：14,16;D12S391：20,21;FGA：23,24.
  9. 9. The lung adenocarcinoma cell line according to any of claims 1 to 6, characterized in that it originates from chinese lung adenocarcinoma patients; Optionally, the cell line is derived from a hydrothorax sample of a patient with chinese lung adenocarcinoma.
  10. 10. The lung adenocarcinoma cell line according to any of claims 1 to 6, wherein the lung adenocarcinoma cell line comprises human lung adenocarcinoma cells SDL, and the preservation number is CCTCC NO: C2025235, the preservation number is 2025, 8 and 13, and the preservation address is the university of Wuhan, hubei province.
  11. 11. A lung adenocarcinoma cell isolated from the lung adenocarcinoma cell line of any one of claims 1-10.
  12. 12. An engineered cell line, wherein the engineered cell line comprises the cell line of any one of claims 1-10; Optionally, the engineered cell line further comprises an exogenous nucleic acid; Optionally, the exogenous nucleic acid comprises at least one of plasmid DNA or at least a portion thereof, siRNA or at least a portion thereof, mRNA or at least a portion thereof, miRNA or at least a portion thereof, viral vector or at least a portion thereof; Optionally, the exogenous nucleic acid comprises at least one of a marker gene or at least a portion thereof, a therapeutic gene or at least a portion thereof, a functional regulatory gene or at least a portion thereof; Optionally, the marker gene comprises at least one of a luciferase gene, a fluorescent protein gene, a β -galactosidase gene; Optionally, the modified cell strain comprises human lung adenocarcinoma cells SDL-luci, and has a preservation number of CCTCC NO: C2025236, and is preserved in China center for type culture Collection, with a preservation date of 2025, 8 months and 13 days, and a preservation address of the university of Wuhan in Wuhan district of Wuhan, hubei province.
  13. 13. A lung adenocarcinoma animal model, characterized in that the tumor of said animal model is formed by the lung adenocarcinoma cell line of any of claims 1-10.
  14. 14. The lung adenocarcinoma animal model according to claim 13, characterized in that it is constructed by inoculating the lung adenocarcinoma cell line according to any of claims 1-10 in an immunodeficient animal.
  15. 15. Use of a lung adenocarcinoma cell line according to any of claims 1-10 or a lung adenocarcinoma animal model according to claim 13 for the preparation of a kit or reagent for the study or screening of a drug, for the study of the mechanism of development of lung adenocarcinoma, or for the construction of an in vitro or in vivo model system for the screening of a drug or the study of the mechanism.

Description

Lung adenocarcinoma cell line, lung adenocarcinoma animal model and application The application claims priority from China patent application filed by the China patent office on the date of 2025, 10 and 21, with the application number 2025115092860 and the application name of "a cell strain derived from EGFR mutant lung adenocarcinoma hydrothorax and luciferase marker strain and application thereof", and the whole content of the cell strain is incorporated by reference. Technical Field The application belongs to the technical field of biology, and particularly relates to a lung adenocarcinoma cell line, a lung adenocarcinoma animal model and application. Background Human Epidermal Growth Factor (EGFR) is one of key driving genes of lung adenocarcinoma, EGFR mutation proportion of patients with Chinese lung adenocarcinoma is high, EGFR-TKI drug resistance and tumor metastasis are the current diagnosis and treatment core dilemma. Human Epidermal Growth Factor (EGFR) is one of key driving genes of lung adenocarcinoma, and currently marketed human epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) can obviously improve prognosis of patients, but most patients inevitably generate drug resistance after a period of treatment in clinical practice, so that treatment failure is caused, meanwhile, tumor metastasis, especially brain metastasis, is taken as another key factor influencing prognosis of lung adenocarcinoma patients, so that the difficulty of clinical treatment is further increased, and the two factors together form a core dilemma to be broken through in the current lung adenocarcinoma diagnosis and treatment field. Lung adenocarcinoma cell lines carrying EGFR mutations commonly used in the current scientific research field, such as NCI-H1975, HCC827, NCI-H1650, NCI-820 and the like, are all derived from European and American populations. Considering that different populations may have significant differences in tumor gene mutation spectra, drug sensitivity and disease progression characteristics, the Europe and America-derived cell lines are difficult to completely simulate the disease characteristics of Chinese lung adenocarcinoma patients, and cannot meet the preclinical research demands such as EGFR-TKI drug resistance mechanisms, transfer rules and the like of Chinese patients. Although the PC9 cell line also carries EGFR mutations and is derived from the eastern asian population (japanese patient), with some similarity to the EGFR mutation background of chinese patients, the cell line is derived from lung cancer in situ tissue, lacks metastatic properties, and has significant limitations in lung adenocarcinoma metastasis-related mechanism studies. To solve the cell line demand of lung adenocarcinoma metastasis research, development attempts aiming at a metastasis characteristic cell strain are already carried out in the prior art, and a high-potential brain metastasis lung cancer cell strain H1975luc is disclosed at presentBM4. The cell strain is obtained by transferring a NCI-H1975 cell line from Europe and America into a luciferase gene, repeatedly inoculating through a left ventricle of a nude mouse, culturing and screening in vitro, and has high brain transfer characteristics, but the NCI-H1975 of the cell strain is not derived from a transfer part of a patient, and the process of constructing a transfer model through artificial left ventricle injection has obvious difference with the actual pathological process of natural infiltration and transfer of cancer cells of the clinical patient, so that the biological characteristics of clinical tumor transfer are difficult to accurately reflect. More importantly, the development of the cell strain does not establish the association between the transfer characteristic and EGFR-TKI drug resistance, can not meet the requirement of the current scientific research field on EGFR-TKI drug resistance and tumor transfer synergistic mechanism research, and greatly limits the application value of the cell strain in lung adenocarcinoma targeted therapy drug resistance and transfer combined research. Disclosure of Invention The application aims to provide a lung adenocarcinoma cell line, a lung adenocarcinoma animal model and application thereof, and aims to solve the problem that a comprehensive model which can be simultaneously used for EGFR-TKI drug resistance mechanism and tumor metastasis mechanism research is lacking in lung adenocarcinoma research of Chinese population. In order to achieve the purposes of the application, the technical scheme adopted by the application is as follows: in a first aspect, the application provides a lung adenocarcinoma cell line carrying a genetic mutation comprising at least one of a clinically significant class I or class II genetic mutation. In some embodiments, the class I gene mutation comprises a deletion mutation that carries the EGFR gene. In some embodiments, the class II gene mutation comprises at lea