| BackgroundEsophageal cancer（EC）is an invasive disease.As one of the most common cancers in the world,the 5-year survival rate is low.The current treatments of EC need to be changed to improve the survival rate of patients.The regulation of transcription factors（TFs）in the gene may be involved in tumor progression and the biology of cancer stem cells（CSCs）.SOX1（sex determining region Y-box protein 1）as a transcription factor can be expressed in a variety of malignant tumors and is closely related to the clinicopathology of tumor patients and may be a potential new therapeutic target.However,there are relatively few studies on the effects of SOX1 and the occurrence and development of esophageal cancer.Therefore,this study mainly explores the effect of SOX1 on the proliferation and invasion of esophageal cancer cells,and analyzes the possible mechanism between SOX1 and Wnt signaling pathway,so as to provide reference significance for the prevention and treatment of esophageal cancer.Objective（1）To detect the effect of SOX1-OE on the proliferation,clone formation,migration and apoptosis of esophageal cancer cells.（2）To study the mechanism of SOX1 on esophageal carcinoma by analyzing the effect of SOX1-OE on Wnt/β-catenin signaling pathway.Methods（1）Construction and transfection of SOX1 overexpression vector.The m RNA sequence of SOX1 was queried by NCBI and synthesized to construct the overexpression vector,and transfected it into ECA109 and EC9706 cells respectively.（2）Effect of SOX1 overexpression on the growth and proliferation of EC cells.The m RNA expression was detected by q PCR after SOX1 overexpression.The proliferation of cells were detected by MTT,crystal violet staining and scratch assay.The proliferation curve was drawn according to the OD value measured by the microplate reader.（3）Effects of SOX1 overexpression on clone formation,migration and apoptosis of EC cells.The ability of cloning to form was judged by the number and size of clones.The ability of cells to migrate was analyzed by observing the scratch assay.The nuclei after staining with Hochest 33342 were observed under fluorescence microscope in order to analyze cell apoptosis.（4）The mechanism of Sox1 overexpression on EC cells.In order to further explore the mechanism of SOX1 as a tumor suppressor,the effect of SOX1 on Wnt/β-Catenin signaling pathway was analyzed by Western blot.Results（1）The SOX1 overexpression vector constructed was transfected into the cells.The high transfection efficiency was observed by green fluorescence.q PCR showed that the expression of SOX1 gene in SOX1-OE group was significantly higher than that in the control group,indicating that SOX1-OE esophageal cancer cell line was successfully established（P < 0.001）.（2）Cell proliferation assay showed that compared with the control group,the OD value of SOX1-OE group decreased significantly after transfection,indicating that the inhibition rate of SOX1-OE group increased and cell proliferation was inhibited.（3）The results of Hochest33342 staining under fluorescence microscope showed that the nuclei of SOX1-OE group became smaller than those of the control group,and some of them appeared obvious shrinkage.（4）Western blot results suggested that the expression of Wnt signaling pathway related genes β-catenin,cyclin D1 and c-Myc decreased significantly,and the expression of GSK-3β increased.ConclusionThe overexpression of SOX1 significantly inhibited the proliferation of EC9706 and ECA109 cells,resulting in the significant decline of cell clone formation and migration ability.At the same time,SOX1-OE can induce the occurrence of apoptosis.Its molecular mechanism may be closely related to the regulation of Wnt/β-catenin signaling pathway. |
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