Discussion And Application Of 3D Culture Model In Lung Cancer Metastasis

In 2020,lung cancer accounted for 11.4% of the global number of new cancers and 18%of the global cancer deaths.Among them,non-small cell lung cancer(NSCLC)accounts for about 80% of all lung cancer.Lung cancer metastasis is the main cause of death in patients with NSCLC.The treatments are limited when lung cancer has distant metastasis(stage IV),and the prognosis is poor.Therefore,it is still necessary to constantly explore the regulatory mechanism of lung cancer metastasis and provide new treatment strategies.At present,the 2D culture model and mouse model of lung cancer metastasis cannot truly reflect the situation of lung cancer metastasis in human body,but 3D culture model can not only simulate the interaction between cells in vivo,but also simulate the hypoxia and necrosis of tumor in vivo.Based on the above reasons,this study used non-small cell carcinoma cell lines A549 and H838 to screen lung cancer metastasis related genes at 3D level.Firstly,in order to simulate the hypoxic state of tumor in vivo,hanging drop culture was used to form cell spheroids;Then,the cell spheroids were injected into the collagen Ⅰ to invade,the invasive cells and spheres of spheroids were separated respectively to circulate this process;Finally,through this process,the cell lines with low invasive ability were screened for sequencing analysis,and the candidate genes were verified at the 3D level,and the correlation between the candidate genes and the prognosis of lung cancer patients was also analyzed.In order to study the regulation mechanism of lung cancer metastasis,this study is committed to establishing a repeatable 3D screening model of lung cancer metastasis,which is of great significance to the research and development of treatment strategy for patients with lung cancer metastasis.The results are as follows:1.A549 and H838 tumor cell spheroids were produced by hanging drop culture.2.A549 cell line with low invasive ability and H838 cell line with high invasive ability were obtained by screening.3.Candidate genes that may affect the invasion of A549 cells at 3D level:KRT4,OTUB2,NTRK3,SLC25A24 and ANKRD35.4.Construct the A549 stable cell lines with KRT4 and OTUB2 knockout,and verify the effects of KRT4 and OTUB2 on the invasion ability of A549 cells at the 3D level.The results showed that KRT4 and OTUB2 significantly inhibited the invasion of A549 cells.