The Mutual Influence And Transformation Between Human Embryonic Lung Fibroblasts And Lung Adenocarcinoma Cancer Cells In Vitro Co-culture

Objective: In this study,human embryonic lung fibroblasts and lung adenocarcinoma cancer cells are co-cultured in vitro to make clear of the interaction and influence between normal fibroblasts and tumor cells in the tumor microenvironment.And immunofluorescence staining reveals that a-SMA in extracellular matrix is obviously up-regulated,that demonstrates normal fibroblasts are activated with specific expression and characteristics of tumor-associated fibroblasts and “activated” fibroblasts could induce tumor cells to undergo changes in their biological behaviors such as proliferation ability,migration ability,invasion ability and sensitivity to conventional chemotherapy drugs.Methods: By studying the interaction and transformation between normal fibroblasts and tumor cells,the in vitro model is applied in the multifactorial interaction process of tumor and matrix environment.1.Human embryonic lung fibroblasts and lung adenocarcinoma cells are in vitro co-culture to clarify the specific overexpression of a-SMA in "activated" fibroblasts by immunofluorescence technique in order to sufficiently prove the possibility of the transition from normal fibroblasts to tumor-associated fibroblasts.2.The cell proliferation assay is used to compare the proliferation ability of induced lung adenocarcinoma cells 3.The flow cytometry detection is used to analyze the distribution of cell cycle and apoptosis.4.The scratch assay and Transwell migration assay are used to compare the migration ability of induced lung adenocarcinoma cells.5.The Transwell invasion assay is used to compare the invasion ability of induced lung adenocarcinoma cells.6.The sensitivity to chemotherapeutic drugs such as cisplatin of induced lung adenocarcinoma cells is detected and compared by MTS assay.Results: Normal fibroblasts can produce interstitial matrix that promotes cell proliferation and specifically express Vimentin proteins,while a-SMA is hardly expressed.The tumor-associated fibroblasts(CAFs)can synthesize and secrete large amount of laminin-1,a-SMA and so on,and the expression of a-SMA is up-regulated with the increase of handling time.1.human embryonic lung fibroblasts and lung adenocarcinoma cells are in vitro co-culture that can activate the normal fibroblasts into tumor-associated fibroblasts(CAFs).And immunofluorescence staining has showed "activated" fibroblasts overexpress a-SMA,which is a specific marker for tumor-associated fibroblasts.After induction,the proliferation ability,migration ability and invasion ability of lung adenocarcinoma cells are significantly enhanced,and the sensitivity to chemotherapeutic drugs such as cisplatin is significantly decreased.Conclusions: 1.The normal fibroblasts induced by tumor cells can be transformed into tumor-associated fibroblasts(CAFs),that have the properties of tumor-associated fibroblasts playing an important role in the tumor occurrence and development.2.Induced tumor cells are significantly enhanced their proliferation ability,migration ability,invasive ability and resistance to the conventional chemotherapy drug.