Preliminary Studies On The Correlation Of β-F1-ATPase And Tumor

F₁F₀-ATP synthase was considered as a protein strictly located in the inner membrane of mitochondria previously. However, current studies have demonstrated that components of F₁F₀-ATP synthase exist on the plasma membrane of tumor cells and adipocytes, which was published as ectopical plasma membrane F₁F₀-ATP synthase. F₁F₀-ATP synthase exist on the surface of many tumor cells, but always with different ratio. It is reported that the functional monoclonal antibody against theβsubunit of F₁F₀-ATP synthase can enhance the cancer and its resistant strains'sensitive to the chemotherapy. This research is aimed at studying the location of theβsubunit of F₁F₀-ATP synthase in tumor cells. At the same time, the effects of the different expression of theβsubunit to cell proliferation has been studied, and the function of F₁F₀-ATP synthase on the surface of tumor cell would be exploited.In order to study the location of theβsubunit of F₁F₀-ATP synthase,βsubunit was cloned to the vector which contains Fluorescent labels. Extract the total RNA from cells, obtain cDNA by reverse transcript and use PCR to amplify atp5b, which was cloned to the Eukaryotic expression vector pEGFP–N₁. After the transfection, fluorescent was observed on cell surface under the fluorescence microscope.To investigate the effects of the different expression of theβsubunit to cell proliferation, we designed three shRNA to interfered the expression of ATP5B. Cell was cultured on normal and hypoxic conditions after transfection, cell proliferation was detected by MTT assays. The results show the proliferation rate of cells transfected with shRNA1 or shRNA2 was more than 1, and cells transfected with shRNA3 was less than 1.