| 罗伊凡,余 力,楼利群,吕 牧,余娟娟,张新宝,张箴波,覃祚树.TFCP2L1在子宫内膜癌细胞增殖及侵袭中的作用[J].,2024,(10):1801-1806 |
| TFCP2L1在子宫内膜癌细胞增殖及侵袭中的作用 |
| Role of TFCP2L1 in Endometrial Cancer Cell Proliferation and Invasion |
| 投稿时间：2024-01-08 修订日期：2024-01-31 |
| DOI：10.13241/j.cnki.pmb.2024.10.001 |
| 中文关键词: TFCP2L1 子宫内膜癌 细胞周期 细胞增殖 细胞迁移 |
| 英文关键词: TFCP2L1 Endometrial cancer Cell cycle Cell proliferation Cell migration |
| 基金项目:国家自然科学基金面上项目（82371642）；中央高校基本科研业务费专项资金资助（22120230312） |
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| 中文摘要: |
| 摘要 目的：研究人子宫内膜癌中TFCP2L1的表达情况以及分析TFCP2L1对子宫内膜癌细胞增殖及迁移能力的影响。方法：（1）通过TCGA 及GTEx数据库分析子宫内膜癌中TFCP2L1的表达水平及患者生存期。采用Western blot验证正常子宫内膜上皮细胞与多种子宫内膜癌细胞系中TFCP2L1的表达情况。（2）使用CRISPR-Cas9技术敲除Ishikawa细胞系的TFCP2L1，并用流式分选技术筛选单个细胞进行培养，形成单克隆细胞系，以此来研究TFCP2L1对子宫内膜癌的细胞周期和细胞增殖的影响。通过 Western blot 及细胞免疫荧光检测细胞周期相关蛋白的表达，检测细胞增殖情况，采用平板克隆实验及CCK8实验。（3）通过Transwell小室及划痕实验对侵袭和转移能力进行检测。结果：TCGA 及GTEx数据库分析发现TFCP2L1在子宫内膜癌中高表达且与肿瘤患者预后不良相关。敲除TFCP2L1后，Ki67、Cyclin D1 和 Cyclin D2的蛋白水平显著下调，CCK8及平板克隆实验结果表明，敲除TFCP2L1能够显著降低子宫内膜癌细胞的增殖能力。划痕实验及Transwell侵袭实验结果表明敲除TFCP2L1的子宫内膜癌细胞侵袭迁移能力均减弱。结论：本研究证明了TFCP21L1是子宫内膜癌的促癌因子。TFCP2L1的高表达可能与子宫内膜癌预后不良相关。敲除TFCP2L1可以抑制子宫内膜癌的侵袭和转移。 |
| 英文摘要: |
| ABSTRACT 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 monoclonal 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 assays. 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. Conclusion: This study demonstrated 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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