The Effects Of ING5 Gene On The Malignant Phenotype Of Breast Cancer Cells

ObjectiveThis study aimed to clarify that the protein of the inhibitor of growth 5(ING5)gene overexpression in human breast cancer cells Bcap-37 influenced cell proliferation,cell cycle,apoptosis,migration,invasion and epithelialmesenchymal transition(EMT),and may influence EMT through PI3K/AKT signal pathway.MethodsAfter stably transfected with eukaryotic p EGFP-N1-ING5 plasmid and GFP-empty plasmid,human breast cancer cells Bcap-37 had the expression of green fluorescent protein measured by fluorescence microscope measured by quantitative real-time PCR,and the high expression of ING5 measured by western blot.Bcap-37-ING5 cells were the experimental group,while Bcap-37-GFP cells for the mock group and Bcap-37 for the control group.In human breast cancer cells Bcap-37,ING5 overexpression were subjected to the measurement of proliferation by MTT,cell cycle and cell apoptosis detected by flow cytometry,cell migration and invasion ability detected by cell wound scratch assay and Transwell experiment.Several EMT associated gene Ecadherin and N-cadherin in the level of m RNA and protein were detected by quantitative real-time PCR and western blot.Constructed nude mouse tumormodel,the effects of ING5 overexpression on Bcap-37 cells in the EMT phenotype were detected by immunohistochemistry.The effects of overexpression of ING5 gene on Bcap-37 cells in the phosphorylation of PI3 K and AKT were detected by western blot.ResultsHuman breast cancer cells Bcap-37 overexpressing ING5 protein with GFP-tag by stable transfection and screening were acquired.Compared with the control group and the mock group,ING5 overexpression decreased cell proliferation rate,caused G2 arrest of Bcap-37 cells,increased cells apoptosis rate,decreased the cell migration and invasion ability.ING5 overexpression might have reverse effect for EMT phenotype by downregulated the level of phosphorylation of PI3 K and AKT in PI3K/AKT pathway(P<0.05).ConclusionsING5 is closely related to pathogenesis,metastasis and differentiation of breast cancer.In human breast cancer cells Bcap-37,ING5 expression might have reverse effects for malignant phenotype of breast cancer cells,which caused low proliferation and G2 arrest of Bcap-37 cells,increased apoptosis,decreased the cell migration and invasion ability.ING5 overexpression might have reverse effects for EMT phenotype by inhibited PI3K/AKT pathway,be employed to indicate the biomarker of prognosis of breast cancer patients and be regarded as a target of gene therapy.