Effects Of CPNE1 On Proliferation And Immigration In Breast Cancer Cells

Purpose: Breast cancer is one of the most prevailing malignancies in women worldwide.As a highly conserved calcium-dependent phospholipid binding protein with functions of regulating signal transduction and membrane transport,copine1(CPNE1)is involved in the carcinogenesis and development of various tumors(including non-small cell lung cancer,osteosarcoma,prostate cancer,and liver cancer,etc.),however its relationship with breast cancer has not been reported.In the presented research,we intend to study the effect of CPNE1 on breast cancer cell proliferation,invasion and migration,and analyze its possible mechanism,providing some new ideas for the understanding of breast cancer.Methods: 1.Database analysis and immunohistochemical techniques were used to investigate the expression differences of CPNE1 in the breast cancer tissues and normal breast tissues(or non-tumor tissues)and their correlation with clinical indexes.2.Western blot and PCR were used to detect the background expression of CPNE1 in breast cancer cells(MCF-7,MDA-MB-231 and HCC1937)and CPNE1 down-regulated cell model was constructed by lentiviral vectors to detect the effects of it on the function of breast cancer.3.MTT,Brdu and clone formation experiment were utilized to detect the changes of cell proliferation ability in CPNE1 down-regulation cell model.4.Flow cytometry was used to investigate the effects of CPNE1 on cell cycle and apoptosis.5.Cell scratch assay and transwell experiments were performed to observe the influences of CPNE1 on the cell migration and invasion.6.Western blot was conducted to explore the expression of related proteins in the downregulated CPNE1 cell model.Results: 1.Indicated by the results of TCGA database analysis and immunohistochemistry,the CPNE1 expression in breast cancer tissue was higher than normal breast tissue(or nontumor tissue),however,there were no significant correlation between the expression of CPNE1 with clinical indexes,such as tumor size(T),lymph node metastasis(N),distant metastasis(M),TNM staging and grade of differentiation.2.Breast cancer cells(MDA-MB-231,HCC1937)had high expression at the protein and m RNA levels.Compared with the vector control group,the cells’ expression of CPNE1 was significantly suppressed after infected with sh RNA.3.The proliferation and colony-forming ability of two breast cancer cells were significantly reduced by the down-regulation of CPNE1 expression.4.Down-regulating the expression of CPNE1 increased the apoptosis rate of two breast cancer cells,and arrested the MDA-MB-231 cell cycle in G1.5.The migration and invasion migration capacity of breast cancer cells were obviously inhibited when the expression of CPNE1 was kept at the low level.6.The expressions of p-AKT was decreased and TP53 was increased by the downregulation of CPNE1,which suggested that CPNE1 might interact with those pathways.Conclusions: 1.The expression of CPNE1 in breast cancer tissues was significantly higher than that in normal breast tissues(or non-tumor tissues);2.The proliferation,migration and invasion of breast cancer cells were enhanced by the expression of CPNE1;3.The function of CPNE1 may be related to AKT and TP53.In summary,CPNE1 may be associated with the development of breast cancer and may be a potential therapeutic target for breast cancer.