Effects Of Coronin 1a On Proliferation And Metastasis Of Breast Cancer Cells

Objective: Coronin protein family was found in the purification of cytoskeletal proteins,and Coronin 1a is one of the most widely studied Coronin proteins.In the cell,Coronin 1a is present as a cofactor in most of the actin-related cytoskeletal motion process.Coronin 1a is also found to be associated with parasite-host reactions,nervous system development,as well as neurological signaling.However,the role of Coronin 1a in tumor development has not been elucidated.Previous studies have shown that Coronin 1a expression in different breast cancer cell lines is quite different,suggesting that Coronin 1a may be associated with different tumor characters in different breast cancer subtypes.This study focused on the relationships between the expression of Coronin 1a and the proliferation and metastasis activities of breast cancer cells,and analyzed the possible pathways.We briefly explain the possible role of Coronin 1a in the cell cycle and EMT process.Content: Analyze the expression of Coronin 1a in different breast cancer cell lines,Screen the cell line of low Coronin 1a expression;synthesize the Coronin 1a overexpressed vector by artificial synthesis;Investigate the changes of the proliferation and metastasis activities of tumor cells after high expression of Coronin 1a and analyze the causes.Investigate the effect of high expression of Coronin 1a on tumorigenesis in nude mice.Method: In this study,we firstly measure the expression of Coronin 1a in all breast cancer cell lines of Key Laboratory of Breast Cancer,Tianjin Medical University Cancer Institute and Hospital.And screened Coronin 1a low expressed cell line.Furthermore,Coronin 1a was amplified by reverse transcription and PCR,and then constructed the eukaryotic expression vector of Coronin 1a by ligating with the pc DNA3.0-HA.Coronin 1a overexpression plasmids were successfully transfected into Coronin 1a cells by using Fu GENE,which successfully constructed the highlyexpressed breast cancer cell lines of Coronin 1a.The relationship between Coronin 1a in breast cancer cell proliferation was investigated by MTT,plate cloning,EDU and flow cytometry.The effects of Coronin 1a in breast cancer cell migration were investigated by Transwell,scratch test and EMT correlation index.Finally,the effectof Coronin 1a on tumorigenic ability of living animals was investigated by subcutaneously injecting cell suspension which highly expressed Coronin 1a into nude mice.In our study,the role of Coronin 1a in the proliferation and metastasis process of breast cancer was discussed for the first time,and the possible pathways of intervention were discussed.Results: In this study,we found that Coronin 1a was low in MDA-MB-231 cell line and SK-Br-3 cell line and was highly expressed in MCF-7 cell line and MCF-10 A cell line,and was negative in T47 D and 293 cell lines.We constructed the Coronin 1a highly expressed cell line by constructing pc DNA3.0-HA-Coronin 1a plasmid and transfected it into MDA-MB-231 and SK-Br-3 cells.MTT,plate cloning and EDU experiments confirmed that overexpression of Coronin 1a significantly promoted the proliferation of breast cancer cells.We use the Flow cytometry to detect the cell cycle,and found that when Coronin 1a was overexpressed,the S phase of the breast cancer cell cycle was significantly increased.Suggesting that Coronin 1a may affect the proliferation of breast cancer cells by regulating the cell cycle.Transwell and scratch tests confirmed that overexpression of Coronin 1a significantly enhanced the ability of breast cancer cells to metastasize.Western blot showed that the expression of E-cadherin was decreased and the expression of N-cadherin,Vimentin and Snail were increased,suggesting that the effect of Coronin 1a on the ability of metastasis of breast cancer cells may be related to the process that Coronin 1a can facilitates the EMT process.Nude mice were confirmed to have a higher tumorigenic ability in breast cancer cells after the overexpression of Coronin 1a,and Ki-67 was significantly higher than that of control group.Conclusion: Coronin 1a is closely related to the proliferation and metastasis of breast cancer.Coronin 1a may regulate the proliferation of breast cancer cells by influencing cell cycle regulation and regulate the ability of breast cancer cell movement by promoting the EMT process.