Experimental Study On The Effects Of Expression Regulation Of Amphiregulin—E-Cadherin On Invasion,Migration,Adhesion And Proliferation Of Human Chondrosarcoma Cells

Objective Chondrosarcoma is the second most common malignant bone tumor after osteosarcoma and is most common in adult males.So far,the diagnosis and treatment of this tumor is still a difficult problem for researchers and clinical workers all over the world.Radiotherapy and chemotherapy are ineffective for most chondrosarcomas,and surgery is by far the most effective treatment.However,patients are often unable to undergo surgery,and surgical treatment can also cause disability or limb dysfunction,so we need to know more about the occurrence and development of chondrosarcoma.E-cadherin is one of the transmembrane glycoprotein cadherins.It frequently appears in the process of epithelial-mesenchymal transition of various cells,and the conversion of E-cadherin to N-cadherin is one of the important characteristics of this process.Amphiregulin is a member of the epidermal growth factor family,which can regulate the invasion,migration and proliferation of different cells,and also plays an important role in the development of various tumors.This study explores the effect and the related mechanisms of amphiregulin on chondrosarcoma,and also involves the process of epithelial-mesenchymal transition of chondrosarcoma cells,which is helpful for human to deepen the understanding of chondrosarcoma,and also has important significance for the future progress of diagnosis and treatment of chondrosarcomas.Methods Chondrosarcoma cell lines SW1353 and CAL-78 were cultured in complete medium with a certain concentration of amphiregulin to obtain chondrosarcoma cells affected by amphiregulin.Endogenous amphiregulin down-regulated cells were obtained by amphiregulin Si RNA transfection,and chondrosarcoma cells were obtained by treating cells with MEK inhibitor U0126 and PI3 K inhibitor LY294002,and these cells were treated with amphiregulin again for the next test.Transwell invasion test,CCK8 test,scratch test and adhesion test were used to measure the invasion,proliferation,migration and adhesion ability of cells in each group.RT-PCR and Western Blot were used to detect the changes of protein and gene levels of E-cadherin,N-cadherin,SLUG and other related factors during the experiment.Results 1.After treatment with amphiregulin,the invasion of SW1353 and CAL-78 chondrosarcoma cells increased,and the expression of E-cadherin decreased,while the expression of N-cadherin and SLUG increased,and the phosphorylation levels of MEK/ERK and PI3K/AKT pathway related proteins increased.2.Chondrosarcoma cells transfected with Amphiregulin Si RNA,MEK inhibitor U0126 and PI3 K inhibitor LY294002 were treated with Amphiregulin again,but the above manifestations were inhibited.3.After treated with amphiregulin,the migration,proliferation and adhesion ability of chondrosarcoma cells were enhanced,and the gene expression of Snail,Twist1 and other important regulatory factors in the process of epithelial-mesenchymal transition were also increased.All the experiments in this study were repeated three times,and the p values of statistical detection were less than 0.05,and the results were significantly different.Conclusions In this study,amphiregulin enhances chondrosarcoma cell invasion by down-regulating E-cadherin expression.MEK/ERK and PI3K/AKT pathways and epithelial-mesenchymal transition may be involved in this process.At the same time,amphiregulin also affected the proliferation,migration and adhesion of chondrosarcoma cells.