Lu Yanyang, Wei Ying, Shen Xiaoqin, Tong Yixi, Lu Jin, Zhang Yahui, Ma Yun, Zhang Rong
Department of Gynecology, The Second Affiliated Hospital of Soochow University, N0.1055, Sanxiang Road, 215000, Suzhou, China.
Genes Genomics. 2023 Nov;45(11):1423-1431. doi: 10.1007/s13258-023-01416-3. Epub 2023 Aug 30.
Endometrial carcinoma (EC) is the most prevalent gynecological cancer. Transcription factor (TF) regulates a large number of downstream target genes and is a key determinant of all physiological activities, including cell proliferation, differentiation, apoptosis, and cell cycle. The transcription factor E2F1 shows prominent roles in EC. BMI1 is a member of Polycomb suppressor Complex 1 (PRC1) and has been shown to be associated with EC invasiveness. It is currently unclear whether E2F1 can participate in the proliferation, migration, and invasion processes of EC cells by regulating BMI1 transcription.
We investigated whether E2F1 could participate in the proliferation, migration, and invasion processes of EC cells by regulating BMI1 transcription, in order to further clarify the pathogenesis and etiology of EC, and provide reference for identifying potential therapeutic targets and developing effective prevention and treatment strategies for this disease.
Human endometrial epithelial cells (hEECs) and human EC cell lines were selected. E2F1 expression was assessed by Western blot. E2F1 was silenced in AN3CA or overexpressed in HEC-1 by transfections, or E2F1 was silenced and BMI1 was overexpressed in AN3CA by cotransfection. Cell proliferation, migration, and invasion were detected by MTT, wound healing, and Transwell assays. The binding sites between E2F1 and BMI1 promoters were predicted through JASPAR website, and the targeted binding was verified by dual-luciferase report and ChIP assays.
E2F1 was up-regulated in human EC cell lines, with its expression highest in AN3CA, and lowest in HEC-1. AN3CA invasion, migration, and proliferation were repressed by E2F1 knockdown, while those of HEC-1 cells were promoted by E2F1 overexpression. E2F1 overexpression increased the activity of wild type BMI1 reporter vector promoter, while this promotion was weakened after mutation of the predicted binding site in the BMI1 promoter. In the precipitated E2F1, BMI1 promoter site level was higher than that of IgG immunoprecipitant. BMI1 silencing suppressed AN3CA cell growth. BMI1 overexpression partially abrogated E2F1 silencing-inhibited EC cell growth.
E2F1 promoted EC cell proliferation, invasion, and migration by promoting the transcription of BMI1.
子宫内膜癌（EC）是最常见的妇科癌症。转录因子（TF）调控大量下游靶基因，是包括细胞增殖、分化、凋亡和细胞周期在内的所有生理活动的关键决定因素。转录因子E2F1在EC中发挥着重要作用。BMI1是多梳抑制复合物1（PRC1）的成员，已被证明与EC的侵袭性有关。目前尚不清楚E2F1是否能通过调节BMI1转录参与EC细胞的增殖、迁移和侵袭过程。
研究E2F1是否能通过调节BMI1转录参与EC细胞的增殖、迁移和侵袭过程，以进一步阐明EC的发病机制和病因，并为确定潜在治疗靶点及制定有效的防治策略提供参考。
选取人子宫内膜上皮细胞（hEECs）和人EC细胞系。通过蛋白质免疫印迹法评估E2F1表达。通过转染使E2F1在AN3CA中沉默或在HEC-1中过表达，或通过共转染使E2F1在AN3CA中沉默且BMI1过表达。通过MTT、伤口愈合和Transwell实验检测细胞增殖、迁移和侵袭能力。通过JASPAR网站预测E2F1与BMI1启动子之间的结合位点，并通过双荧光素酶报告基因和染色质免疫沉淀实验验证靶向结合。
E2F1在人EC细胞系中上调，在AN3CA中表达最高，在HEC-1中最低。E2F1敲低抑制了AN3CA的侵袭、迁移和增殖，而E2F1过表达促进了HEC-1细胞的侵袭、迁移和增殖。E2F1过表达增加了野生型BMI1报告载体启动子的活性，而在BMI1启动子中预测的结合位点突变后，这种促进作用减弱。在沉淀的E2F1中，BMI1启动子位点水平高于IgG免疫沉淀剂。BMI1沉默抑制了AN3CA细胞生长。BMI1过表达部分消除了E2F1沉默对EC细胞生长的抑制作用。
E2F1通过促进BMI1转录促进EC细胞增殖、侵袭和迁移。
