Study On Molecular Mechanisms Of Stromal Cells In Microenvironment For Lung Cancer And Metastasis Woth Bionic Organs-on-microfluidic Chips

BackgroundLung cancer metastasis, which involves a series of complicated pathological process, is closely related to the tumor microenvironment. The tumor microenvironment constitutes the tumor cells, stromal cells and extracellular matrix. On the one hand, the microenvironment promotes lung cancer cells metastasis; on the other hand, lung cancer cells in turn affect the microenvironment. To confirm the molecular mechanisms of lung cancer and its metastasis impacting on microenvironment, will cut off the underlying pathways which involved in the improvement of seeds on the soil and thus may provide a novel therapeutic target for lung cancer patients. But now it is difficult to mimic the real lung tumor microenvironment depending on the existing research techniques, also the reliability needs to be improved. Thus, there is an urgent need to develop a reliable and efficient in vitro culture model to closely mimic the in vivo microenvironment of lung cancer metastasis. Animal testing will be used to qualify the above results gained from the bionic microfluidic lung cancer metastasis model. Fortunately, the developed microfluidic chip technology can meet the above requirements, is expected to provide reliable technical supports for the test, as a new technology platform.Herein, the project intends to construct a bionic microfluidic chip lung physical model, a bionic microfluidic chip lung cancer model to investigate the functional changes of main stromal cells fibroblast submerged in the tumor metastasis microenvironment by means of the superiority of microfluidic chip high throughput and integration and further clarify the related molecular mechanisms of lung cancer metastasis impacting on microenvironment. What’s more, the results are expected to find new targets for the control and treatment of lung cancer metastasis, and to explore the applications of the microfluidic chip and engineering technology for lung cancer.This study contains the following three parts.Methods1. Lung cancer on a microfluidic chip constructionA lung-on-a-chip system was constructed an environment to mimick the in vivo growth conditions of lung cancer cells in vitro by gas-liquid two-phase continuous dynamic cell culture. Based on the above new microfluidic chip platform, lung cancer on a microfluidic chip were constructed to simulate lung cancer formation, invasion and metastasis process and the tumor microenvironment. It provides a technology platform for the subsequent molecular mechanisms of lung cancer and microenvironment.2. Application of Proteomics Technique to screen key molecule for interactions of lung cancer and microenvironmenti TRAQ technology was to screen differential expression of proteins of lung cancer cells and fibroblasts; key molecules were screening through protein interaction network data, statistical enriched KEGG pathways, GO-BP enrichment map information, and so on; It provides a clear basis for possible molecular mechanisms of lung cancer and microenvironment.3. Validation studies on key molecular for interactions of lung cancer and microenvironmentThe function of PPARD, STC-1 interactions of lung cancer and microenvironment with the bionic chip microfluidic platform and conventional platforms.It provide new ideas for microfluidic chip technology and proteomics technologies in the field of cancer biology.Results1. Lung cancer on a microfluidic chip constructionA lung-on-a-chip system was constructed an environment to mimick the in vivo growth conditions of lung cancer cells in vitro. The human bronchial epithelial and microvascular endothelial cells were viable for prolonged periods by gas-liquid two-phase continuous dynamic culture and stromal cells were co-cultured. The results found that the gas-liquid two-phase training mode was suitable for cell growth, and simulate the main physiological function respiratory rhythm and blood exchange; on this basis, lung cancer on a microfluidic chip was constructed to simulate the microenvironment and specific markers of lung cancer cells and fibroblasts of the different stages in the process of lung cancer information, invasion and metastasis.2. Application of Proteomics Technique to screen key molecule for interactions of lung cancer and microenvironmenti TRAQ technology was to screen differential expression of proteins of lung cancer cells and fibroblasts; key molecules were screening through protein interaction network data, statistical enriched KEGG pathways, GO-BP enrichment map information, and so on; Key molecules of lung cancer cells were: PPARD、STC1、ICAM1、F13A1、KRT85、G6PD、RRM2、FLNB、DOCK1、MAGED1, and fibroblasts : PPARD、UBA52、HIST1H2BD、MTHFD1、RPLP2、TUBB6、UBE2C、SORBS2、KRT1、CD2AP. It provides a clear basis for possible molecular mechanisms of lung cancer and microenvironment.3. Validation studies on key molecular for interactions of lung cancer and microenvironmentThe function of PPARD, STC-1 interactions of lung cancer and microenvironment with the bionic chip microfluidic platform and conventional platforms.The results indicated that EMT and invasion of lung cancer cells were downregulated with Si RNA of PPARD, STC-1. It provides new ideas for microfluidic chip technology and proteomics technologies in the field of cancer biology.Conculsions1. The project constructed a lung-on-a-chip and lung cancer-on-a-chip system to mimick the in vivo growth conditions of lung cancer cells in vitro by gas-liquid two-phase continuous dynamic cell culture to simulate lung cancer formation, invasion and metastasis process and the tumor microenvironment. It provides a technology platform for the subsequent molecular mechanisms of lung cancer and microenvironment.2. Application of Proteomics Technique to screen key molecule for interactions of lung cancer and microenvironment. It provides a clear basis for possible molecular mechanisms of lung cancer and microenvironment.3. The function of PPARD, STC-1 interactions of lung cancer and microenvironment with the bionic chip microfluidic platform and conventional platforms. The results indicated that EMT and invasion of lung cancer cells were downregulated.4. It provides a technology platform for the subsequent molecular mechanisms of lung cancer and microenvironment and new ideas for microfluidic chip technology and proteomics technologies in the field of cancer biology.