The Influence Of Interfere FANCF Gene On Biological Characteristic In Breast Cancer Cells

FA/BRCA pathway participate in cell cycle and apoptosis control, regulation of detoxification, survival signal transduction and DNA repair. Disruption of this pathway results in chromosome instability and hypersensitivity to DNA cross-linking agents. Stabilization of FA complex and monoubiquitination of FANCD2 are the critical events and FANCF plays an extremely vital role as a key factor. Low expression and loss function of FANCF could interfere in the FA/BRCA pathway and participate in tumorigenesis and development of drug resistance. At present, It has been confirmed that FANCF is related to development and drug resistance of many kinds of tumors. But there are no reports about whether FANCF participate in development and drug resistance of breast cancer. We investigate effect of FANCF gene silence on biological character in breast cancer cell and whether the regulation of FANCF in breast cancer cell have difference between ER(+) MCF-7 cell and ER(-) 435S cell.MethodsConstrution of breast cancer-FANCF-RNAi cell model, extract FANCF-shRNA plasmid and transfect into MCF-7 and 435S cells, detect mRNA and protein expression of FANCF by RT-PCR and Western blot to confirm whether FANCF gene silence model were established successfully; After FANCF gene silence, investigate the proliferation by MTT assays, the cell cycles and cell apoptosis are measured by flow cytometry after the cells staining by PI only and Annexin-V and PI respectively, also the ability of cell migration and invasion are detected by scratch method and transwell chamber, the ability of DNA injury breakage were measured by comet assays. Successful establishment of MCF-7-FANCF-RNAi,435S-FANCF-RNAi cell model:Compared to Control-shRNA group, mRNA and protein expression of FANCF gene are reduced in FANCF-shRNA group after transfection for 48h,72h in two cells (P<0.05); Effect of FANCF gene silence on biological characteristic of breast cancer cells:FANCF gene silence could inhibit cells proliferation (P<0.05); decrease cells migration and invasion potential (P<0.05); increase the percentage of cell cycle in S-phase (P<0.05) and apoptosis rate (P<0.05); increase DNA injury breakage (P<0.05); The regulation of FANCF in breast cancer cells have little difference between MCF-7 and 435S cells (P>0.05).ConclusionFANCF gene silence could inhibit cells proliferation and invasion potential, increase cells apoptosis and arrest cells in S phase; The regulation of FANCF in breast cancer cells have little difference between MCF-7 and 435S cells.