br for preventing endometrial cancer and improve progestin b
for preventing endometrial cancer and improve progestin-based con-servative treatment in early stage endometrial cancer.
The conception and design of the study: Xiaojun Chen and Congjian
The acquisition of data: Qiaoying Lv, Liying Xie, Yali Cheng, Weiwei Shan, Chengcheng Ning, Bingying Xie, Bingyi Yang, Xuezhen Luo, Qizhi He, Qin Zhu. Analysis and interpretation of data: Qiaoying Lv, Yali Cheng, Yingli Zhang, Zhenbo Zhang, Chenji Wang.
Drafting the article: Qiaoying Lv, Yue Shi and Liying Xie.
Revising it critically for important intellectual content: Xiaojun Chen, Zhenbo Zhang, Chenji Wang.
Final approval of the version to be submitted: Xiaojun Chen and Congjian Xu.
Declarations of interest
This work was supported by National Natural Science Foundation of China (Grant No.81671417 and 81370688), Shanghai Medical Center of Key Programs for Female Reproductive Diseases (2017ZZ010616), Shanghai Science and Technology Development medical guide project (Grant No.17411961000), Municipal Human Resources Development Program for Outstanding Leaders in Medical Disciplines in Shanghai (Grant No. 2017BR035) and Open Research Fund of State Key Laboratory of Genetic Engineering, Fudan University (No. SKLGE-1808) and National Natural Science Foundation of China (Grant No. 81602267).
We also thank Shimin Zhao Lab and Yao Li Lab (School of Life
Fig. 7. Overexpression of A20 increases sensitivity of EC N-octanoyl-L-Homoserine lactone to estrogen via regulation of ERα. A-B. A20 WT promoted EC cell proliferation in the presence of E2, but A20 MT had no such eﬀect. EC cells were treated with 100 nM E2 for 48 h after transfection with A20 plasmids or empty vector for 8 h. A. EdU staining was performed to evaluate cell mitosis. Cells stained with EdU (red fluorescence) were in the S phase of mitosis, and all cells were stained with Hoechst (blue fluor-escence) to detect nuclei. Representative images (left column) and percentage of Edu-positive EC cells (right column) were presented. WT, wild type; MT, mutant type, C103A; Scale bars, 100 μm. B. CCK8 assay was performed to evaluate cell proliferation. C. A20 WT inhibited EC cell apoptosis in the presence of E2, but A20 MT had no such eﬀect. HEC-1A cells were initially transfected with A20-expressing plasmids or empty vectors for 8 h and then treated with E2 for 48 h. Flow cytometry was performed to evaluate the percentage of apoptotic cells in HEC-1A cells. D. A20 upregulated ERα expression in the absence or presence of E2, but A20 MT had no such eﬀect. EC cells were treated with 100 nM E2 for 48 h after transfection with A20-expressing plasmids or empty vectors for 8 h. Cells were harvested for western blotting analysis. E. A20 dose-dependently stabilized ERα protein in the presence of E2. HEK-293T cells were transfected with A20-expressing plasmids or empty vectors for 24 h and then were treated with E2 or ethanol vehicle (EtOH) for 24 h *P < 0.05, **P < 0.01, ***P < 0.001.
Sciences, Fudan University) for their support. We also thank H. Nikki March, PhD, from Liwen Bianji, Edanz Editing China (www.liwenbianji. cn/ac), for editing the English text of a draft of this manuscript.
Appendix A. Supplementary data
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