Role of TFCP2L1 in Endometrial Cancer Cell Proliferation and Invasion
Objective:To investaigate the expression of TFCP2L1 in human endometrial cancer and to analyze the effect of TFCP2L1 on the proliferation and migration of endometrial cancer cells.Methods:(1)The expression of TFCP2L1 in endometrial cancer and the overall survival of patients were analyzed by TCGA and GTEx databases.Western blot was used to verify the expression of TFCP2L1 in normal endometrial epithelial cells and different endometrial cancer cell lines.(2)TFCP2L1 was knocked down in Ishikawa cell line using CRISPR-Cas9,and individual cells were screened and cultured by Fluorescence activated Cell Sorting to form a mono-clonal cell line,in order to investigate the effect of TFCP2L1 on the cell cycle and cell proliferation in endometrial cancer.The levels of cell cycle-related proteins were assessed by Western blotting and immunofluorescence.The proliferation of cells was evaluated using plate cloning assay and CCK8 assay.(3)The invasion and metastasis abilities of the cells were examined by Transwell and scratch as-says.Results:A higher expression level of TFCP2L1 in endometrial cancer and lower overall survival rates in patients with elevated TFCP2L1 levels were indicated by analysis of the TCGA and GTEx databases The protein levels of Ki67,Cyclin D1 and Cyclin D2 were significantly down-regulated,and the protein level of P21 was significantly increased after knockdown of TFCP2L1.Functional assays demonstrated that TFCP2L1 depletion significantly reduced the proliferation of endometrial cancer cells,as evidenced by the CCK8 assay and smaller cell clone sizes observed in plate cloning experiments.The results of scratch assay and Transwell invasion assay showed that the invasive and migratory ability of endometrial cancer cells knocked down TFCP2L1 was reduced.Conclusions:This study demonstrat-ed that TFCP21L1 is a pro-cancer factor in endometrial cancer.High expression of TFCP2L1 may be associated with poor prognosis of endometrial cancer.Knockdown of TFCP2L1 could inhibit the invasion and metastasis of endometrial cancer.
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