Study On Molecular Mechanisms Of Lymphocyte And Tumor Growth-Induced Solid Stress In Microenvironment For The Progression Of Clear Cell Renal Cell Carcinoma With Mechanical Device

The tumor microenvironment is formed of cancer cells,stromal cells and extracellular matrix.The oncogenesis and metastasis of clear cell renal cell carcinoma(ccRCC)is closely related to the tumor microenvironment.However,now most studies are focus on biochemical factors,ignore the possible role of biomechanical factors in human tumors.Therefore,we construct a mechanical device with biochemical stimulus,to further mimic the real renal tumor microenvironment,clarify the related molecular mechanisms of biochemical factors and biomechanical factors for clear cell renal cell carcinoma.The project intend to construct a physical model to study the molecular mechanisms of lymphocyte and tumor growth-induced solid stress in microenvironment for the progression of clear cell renal cell carcinoma.Moreover,we suggest to explore new direction for renal cancer therapy.Methods1.Application of PCR Array to screen key component in microenvironment and its role for renal cancerDepend on self-designed PCR array associated tumor environment,we screened key component CD4 between normal kidney tissue and renal cancer tissue;Key component CD4 was identified by immunohistochemical staining,survival curve,co-focus and functional assay.It identifies key component effected the progression of renal cancer and provides a basis for possible molecular mechanisms.2.In mechanical platform to screen key molecular with the role of biochemical and biomechanical factors for clear cell renal cell carcinoma through gene expression arrayWe constructed tumor microenvironment model with mechanical device to investigate the role of mechanical factor for renal cancer through gene expression array.Based on the platform,we added biochemical stimulus IL-6 to study the progression of tumor cells.It provide a technology platform for the further molecular mechanisms of the effect of microenvironment for clear cell renal cell carcinoma3.Validation studies on key molecular and signal pathway for the progression of clear cell renal cell carcinomaThe function of ?-catenin,CD44 and their regulated signal pathway in clear cell renal cell carcinoma with mechanical platform were determined,It provides evidence for new stress-reducing and IL-6 targeting strategies in cancer therapy.Results 1.Application of PCR Array to screen key component in microenvironment and its role for renal cancerDepend on self-designed PCR array associated tumor environment,we screened CD4 and components associated with epithelial to mesenchymal transition between normal kidney tissue and renal cancer tissue.Further we found the number of ?-SMA(+)cells and CD4(+)cells in both Fuhrman I tissues and Fuhrman III-IV samples was significantly increased compared with the normal kidney tissues,moreover,CD4 expression positively correlated with ?-SMA expression in clear cell renal cell carcinoma.In the next step,we investigated the association between CD4+ T cells and ?-SMA+ cells in clear cell renal cell carcinoma.We evaluated the production of soluble mediators by CD4+ T lymphocytes co-cultured with tumor cells,among them,IL-6 had the highest concentration.After cancer cells were exposed to IL-6,a near complete loss of the epithelial feature occurred,moreover,IL-6 enhancedmigratory and invasive capability of tumor cells.It identifies key component effected the progression of renal cancer and provides a basis for possible molecular mechanisms.2.In mechanical platform to screen key molecular with the role of biochemical and biomechanical factors for clear cell renal cell carcinoma through gene expression arrayTumor microenvironment is not only include biochemical factors but also biomechanical factors.We construct a mechanical device with biochemical stimulus,to further mimic the real renal tumor microenvironment.Cell motility assay and cell viability assay were conducted to evaluate the range of compressive stress.Based on these data,IL-6 treated cells,compressed cells and combined group cells were subjected to gene array analysis and further GO(Gene Ontology)results.Key molecular was ?-catenin,It provides a technology platform for the further molecular mechanisms of the effect of microenvironment for clear cell renal cell carcinoma 3.Validation studies on key molecular and signal pathway for the progression of clear cell renal cell carcinomaThe function of ?-catenin was studied in mechanical device.We found ?-catenin played a critical role in epithelial to mesenchymal transition and cancer stemness.In addition,biochemical factors and biomechanical factors effected the progression of clear cell renal cell carcinoma.It provides evidence for new stress-reducing and IL-6 targeting strategies in cancer therapy.Conclusion 1.Depend on self-designed PCR array associated tumor environment,we screened key component CD4 between normal kidney tissue and renal cancer tissue;Key component CD4 was identified by immunohistochemical staining,survival curve, co-focus and functional assay.It identifies key component effected the progression of renal cancer and provides a basis for possible molecular mechanisms.2.We constructed tumor microenvironment model with mechanical device to investigate the role of mechanical factor for renal cancer through gene expression array.Based on the platform,we added biochemical stimulus IL-6 to study the progression of tumor cells.It provide a technology platform for the further molecular mechanisms of the effect of microenvironment for clear cell renal cell carcinoma3.The function of ?-catenin was studied in mechanical device.We found ?-catenin played a critical role in epithelial to mesenchymal transition and cancer stemness.