The Role Of Magnesium Transporter Subtype 1(MAGT1) In Theprogression And Prognosis Of Colorectal Cancer And The Study On Its Molecule Mechanism

Background and Objective:Colorectal cancer(CRC)is the third largest malignancy in the world and the fourth leading cause of cancer death.Due to the limitations of conventional biomarkers,it is urgent to find new and effective biomarkers to predict the progression and prognosis of CRC.In recent years,researchers have found that magnesium ion(Mg2+)homeostasis is prevalent in tumor cells,and the loss and supplement of magnesium ion can affect the occurrence and development of tumors.Magnesium ion homeostasis is regulated by a variety of transporters,and the research reports on the imbalance of magnesium ion homeostasis caused by the expression change of magnesium ion transporters and the influence on the occurrence,development and prognosis of tumors show an increasing trend year by year.The purpose of this study was to explore the expression of magnesium ion transporter protein magnesium transporter subtype 1(MAGT1)in colorectal cancer and its clinical significance,and to further analyze the regulatory effect and pathogenesis of MAGT1 on the malignant biological phenotype of colorectal cancer cellsMethods:1?Through statistical analysis of GEO database data,the expression of MAGT1 in CRC samples and the clinical information related to the progression and prognosis of colorectal cancer were determined.Fifty-one clinicopathological samples were collected,and the differential expression of MAGT1 in colorectal cancer and para-carcinoma tissues was detected by real-time quantitative polymerase chain reaction(QRT-PCR);The differential expression of 8 types of colorectal cancer cells and normal colorectal epithelial cells was detected by western blot analysis.2?SW620 and HCT116 of high expression MAGT1 colorectal cancer cells were selected,and transient cell strain was constructed by transfection with small interfering RNA and stable cell strain was constructed by transfection with shRNA lentivirus,which were divided into control group and down-regulated group.QRT-PCR and western blot analysis were used to verify the down-regulated efficiency;CCK8 proliferation assay was used to detect changes in cell proliferation in vitro;Using subcutaneous tumor formation assay in nude mice to detect changes in cell proliferation in vivo;Using scratch healing assay to detect changes in cell migration;Using the transwell assay to detect changes in cell invasion;Cell apoptosis and cycle changes were detected by flow cytometry.3?The activation of EGFR signaling pathway was induced by the addition of appropriate epidermal growth factor(EGF)in SW620 and HCT116 stable strains,and the relative expression changes of EGFR signaling pathway related proteins(P-AKT,AKT,P-MEK,MEK,P-ERK,ERK,etc.)in the control group and the down-regulated group were detected by western blot analysis.Results:1?MAGT1 was highly expressed in both human colorectal cancer tissues and cells,and was associated with clinicopathological features and treatment prognosis of colorectal cancer,with statistically significant differences(P<0.05).(1)Statistical analysis of GEO database chip data showed that high expression of MAGT1 was associated with clinicopathological features and chemotherapy prognosis of patients.(2)The results of 51 collected clinical samples showed that MAGT1 was up-regulated in colorectal cancer tissues.(3)MAGT1 was up-regulated in 6 colorectal cancer cells compared with normal intestinal epithelial cells.2?Down-regulation of MAGT1 expression could inhibit the proliferation,migration and invasion of colorectal cancer cells inside and outside the body,and promote cell apoptosis.All the experimental results showed statistically significant differences(P<0.05).(1)The results of CCK8 cell proliferation assay indicated that the proliferation capacity of the down-regulated group decreased compared with the control group In vitro;The results of subcutaneous tumor formation assay in nude mice indicated that the proliferation ability of the down-regulated group decreased compared with the control group in vivo.(2)The results of the scratch healing assay and transwell assay indicated that the down-regulated group had a lower ability of cell migration and invasion than the control group.(3)The results of flow cytometry analysis indicated that the apoptosis rate of the down-regulated group was higher than that of the control group3?Down-regulation of MAGT1 expression inhibited the activation of EGFR signaling pathway and the expression of pathway related proteins,with statistically significant differences(P<0.05).(1)The results of western blot indicated that the relative expressions of P-AKT,P-MEK and P-ERK decreased significantly in the down-regulated group compared with the control group,while the relative expressions of AKT,MEK and ERK did not change significantly.Conclusion:MAGT1 is up-regulated in human colorectal cancer tissues and cells.By regulating the EGFR signaling pathway,highly expressed MAGT1 can lead to the proliferation,migration,invasion,anti-apoptosis and other malignant cell biological phenotypes in colorectal cancer cells,which may be an effective biomarker to predict the progression of colorectal cancer and the prognosis of treatment as well as a potential therapeutic target for patients with anti-EGFR antibody resistance.