Molecular Mechanism And Expression Of VRK1 And The Downstream Gene BANF1 In Esophageal Cancer

ObjectiveTo investigate the effect of the VRK1 and the downstream gene BANF1 on the cell cycle and invasion of esophageal squamous cell carcinoma cell and explore the molecular mechanism that might be involved in the pathogenesis.MethodsVRK1 and BANF1 protein levels were assessed immunohistochemically in esophageal cancer specimens.The clinical characteristics of the esophageal cancer patients were catalogued,and the intensity of VRK1 and BANF1 staining in each sample was assigned a score of 0(negative staining),1(<5% staining),2(<25%staining)or 3(25 – 50% staining).To identify the role of VRK1 and BANF1 in esophageal cancer tissues,VRK1 and BANF1 m RNA levels were examined via real-time PCR in several paired tumor tissues and adjacent normal tissues.We transiently silenced VRK1 and BANF1 expression in EC109 and EC1 cell lines,after which we measured cell proliferation using CCK-8 assays.Western blotting showed that VRK1 and BANF1 were effciently knocked down by transfection of a targeted siRNAResults1.VRK1 and BANF1 are over-expressed in poorly differentiated esophageal cancer biopsies compared with adjacent normal tissues.In the 120 normal esophageal tissues,positivity for VRK1 and BANF1 was observed in 19 cases and 16 cases,respectively,and the corresponding rates of positivity were 15.8% and 13.3%.In the85 cases of well-differentiated esophageal cancer,64 and 61 cases were positive for VRK1 and BANF1,respectively,and corresponding rates of positivity were 75.3%and 71.8%.In the 35 cases of poorly differentiated esophageal cancer,the numbers of VRK1-and BANF1-positive cases were 33 and 32,respectively,and the corresponding rates of positivity were 94.3% and 91.4%.2.VRK1 and BANF1 mRNA expression levels in tumor tissues and adjacent normal tissues shows that they were higher in tumor tissues than in adjacent normal tissues.3.Western blot showed that siVRK 571 was the most effective sequence and VRK1 was efficiently knocked down in both cells,VRK1 depletion decreased expression of BANF1.CCK-8 test showed Depletion of VRK1 that using siRNA suppressed the growth of EC109 and EC1 cells significantly.The results of flow cytometry showed that the cell cycle was blocked after S,cells in S had increased,division in M was decreased,and the difference was statistically significant.Conclusion1.VRK1 and BANF1 are up-regulated in esophageal cancer and that their increased expression is associated with a poor prognosis.VRK1 and BANF1 expression might play an important role in the progression of esophageal squamous cell carcinoma.2.VRK1 depletion may suppress BANF1 expression.The regulatory pathway related to VRK1 and BANF1 promote cell proliferation in esophageal cancer cell lines3.High VRK1 and BANF1 expression could potentially serve as an indicator of a poor prognosis and/or a therapeutic target in esophageal cancer.