Characterization of lung adenocarcinoma cell markers and sensitivity to epidermal growth factor receptor targeting drugs in an organoid culture system
Objective To characterize the structure and related markers of lung cancer-like organs formed by lung adenocarcinoma cells and to conduct a preliminary study on their sensitivity to epidermal growth factor receptor(EGFR)-targeted drugs.Methods Tumor organoid cultures of the commonly used lung adenocarcinoma cell lines A549(EGFR wild-type)and H1975(EGFR T790M/L858R)were per-formed to observe the formation,morphology,and proliferation of lung cancer organoids.The expression of TTF-1,CK7 and Ki-67 was detected by immunohistochemistry and compared with the traditional two-di-mensional culture.The spatial distribution and expression of N-cadherin in A549 and H1975 organoids were detected by immunofluorescence at the same time.The drug sensitivity tests of the first and third gen-eration targeted drugs gefitinib and osimertinib were carried out on lung cancer organs formed by A549 and H1975.The ATP was quantitatively analyzed by 3D-glo method.The data were analyzed by non-linear fit-ting curve with GraphPad Prism9 using the drug action dose as the abscissa and the cell viability value as the ordinate.Student's t test test was used for data comparison between two independent samples,and ANOVA between multiple-group.Results Lung adenocarcinoma cells A549 and H1975 formed organoid spheres with an average diameter of 50 µm at 7 and 5 days,respectively.HE staining showed microstruc-tures,and two-dimensional lung cancer cells did not express the same markers as the organoids they formed.After targeted drug treatment,A549 organs were not sensitive to gefitinib and osimertinib,while H1975 lung cancer organs were resistant to gefitinib and sensitive to osimertinib.The A549 organoid half maximal inhibitory concentration(IC50)was significantly higher in gefitinib than H1975(1.08 µmol/L vs.0.50 µmol/L,t=9.90,P＜0.05).The A549 IC50 was significantly higher in osimertinib than H1975(1.04 μmol/L vs.0.01 µmol/L,t=26.78,P＜0.05).Conclusion Lung adenocarcinoma organoids formed by H1975(T790M/L858R)mutation showed high sensitivity to the three-generation TKI drug osim-ertinib compared with wild-type A549.
Tumor organoidsEpidermal growth factor receptorAdenocarcinoma of lungTumor markersDrug sensitivity
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