Biological Function Of CLDN1 In Esophageal Cancer Cells

Objective: To detect the biological function of CLDN1 in esophageal cancer cells by downregulating the expression of CLDN1 in TE-11 and upregulating the expression of CLDN1 in TE-10.To detect the intracellular distribution of CLDN1 in normal esophageal epithelial cells and tumor cells by western blot and flow cytometry.To scrneen the downstream regulation genes of CLDN1 by gene chip.Methods: Culturing normal esophageal epithelium cell and esophageal cancer cell lines Eca109,Eca9706,TE-1,TE-10,TE-11,Kyse150 in vitro,Lentivirus containing the sequence of CLDN1 shRNA was constructed and transfected into TE-11 cells,and lentivirus of overexpression of CLDN1 was constrcted and transfected into TE-10.Detecting the transfection efficiency by immunofluorescence microscopy and detecting the expression of CLDN1 by western blot after transfection.The intracellular distribution of CLDN1 was determined by western blot and flow cytometry.Cell proliferation ability was detected by cck-8,transwell was used to determine the invasion and migration ability of cells.TE-11 cells were treated with Rhodamine phalloidin staining and subjected to a laser scanning confocal microscope for the observation of cytoskeleton changes.Inoculating the interference group and control group cells of TE-11 and TE-10 in nude mice axillary subcutaneous,observing the tumor volume after 4 weeks.Inoculating intraperitoneally the interference group and control group cells of TE-11 and TE-10 in nude mice,observing the number of celiac planting tumor after 4 weeks.Inoculating the interference group and control group cells of TE-11 and TE-10 in tail vein,observing the number of pulmonary metastasis tumor after 4 weeks.To scrneen the downstream regulation genes of CLDN1 by gene chip after upregulating the expression of TE-10,predicting the downstream pathways related to CLDN1.Results: After downregulating the expression of CLDN1 of TE-11,the proliferation,invasion and migration ability was declined obviously.Laser confocal microscope shows that the cytoskeleton protein fluorescence intensity deceased obviously.The tumor volume and numbers reduced obviously compared with control group.After upregulating the expression of CLDN1 of TE-10,the proliferation,invasion and migration ability was increased obviously.Laser confocal microscope shows that the cytoskeleton protein fluorescence intensity increased obviously.The tumor volume and numbers increased obviously compared with control group.western blot and flow cytometry shows that CLDN1 is mainly distributed in the nucleus of TE-11 cells,and mainly distributed on the membrane of normal esophageal epithelium cells.Gene chip screened 1311 differently expressed genes,and upregulated genes have 626(47.75%),downregulated genes have 685(52.25%).Conclusion: CLDN1 has a carcinogenesis effect in esophageal cancer cells,and mainly distribution in nuclear,which is expected to become an important biomarker for ESCC.