AT1-R在雌激素诱导子宫内膜癌细胞增殖中的作用

潘卓; 杨清; 苏卿

doi:10.3969/j.issn.1000-8179.2011.02.001

摘要: 目的：探讨血管紧张素受体1 （angiotensin receptor 1， AT1-R）在雌激素诱导子宫内膜癌细胞HEC-1A增殖、细胞周期转化中的作用及其对细胞外调节蛋白激酶（extracellular signal-regulated kinases 1/2，ERK1/2）表达的影响。方法：免疫荧光技术检测AT1-R在HEC-1A细胞中的表达；脂质体介导AT1-R-siRNA质粒转染人子宫内膜癌细胞HEC-1A，经G418选择培养，采用Western blot检测转染前后HEC-1A细胞中AT1-R蛋白的表达。MTT法检测无雌激素诱导及雌激素诱导10 min转染前后细胞的增殖；流式细胞技术检测细胞周期； Western blot检测ERK1/2蛋白及ER蛋白表达水平。结果：转染AT1-R-siRNA-GFP后，与未转染组相比AT1-R蛋白表达降低41.79（P<0.01）；沉默AT1-R能够抑制17β-E2对HEC-1A细胞的促增殖作用。17β-E2处理10 min后， HEC-1A细胞G1期细胞减少， S期细胞增多，细胞增殖明显（P<0.05）；AT1-R沉默后，能够抑制17β-E2诱导的HEC-1A细胞周期转化，使G1期细胞增加，S期细胞减少（P<0.05）；雌激素处理10 min后，HEC-1A细胞ERK1/2蛋白表达水平升高，沉默AT1-R后ERK1/2蛋白表达水平又明显降低。17β-E2诱导及AT1-R沉默对HEC-1A细胞ER蛋白表达无明显影响。结论：AT1-R在雌激素诱导子宫内膜癌细胞HEC-1A增殖，细胞周期转化中具有重要作用，其机制可能与ERK1/2表达降低有关，与ER表达无关。

Abstract: The Role of Angiotensin II Type 1 Receptor in Estrogen-induced Proliferation of EndometrialCarcinoma Cell Line HEC-1AZhuo PAN，Qing YANG, Qing SUCorrespondence to: Qing YANG, E-mail: yangq@sj-hospital.orgDepartment of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, ChinaThis work was supported by Shenyang Science and Technology Plan Projects（No. 071219）and Liaoning Bai Qian WanTalents Program (No. 2008921066)Abstract Objective: To investigate the role of angiotensin Ⅱ type 1 (AT1-R) in estrogen-induced proliferation, cell cy-cle progression and expression of ERK1/2 in endometrial carcinoma cell line HEC-1A. Methods: The expression of AT1-Rwas assessed by immunofluorescence. AT1-R siRNA was transfected into human endometrial carcinoma cell line HEC-1Avia Lipofectamine 2000. After screening the cultures with G418, the expression of AT1-R protein was examined by West-ern blot before and after transfection. The effect of AT1-R silencing on 17β-E2-induced proliferation of HEC-1A cells wasmeasured by MTT assay, the cell cycle was analyzed by flow cytometry, and the expression of ERK1/2 protein was exam-ined by Western blot. Results: After transfection with AT1-R siRNA plasmid, the expression of AT1-R protein was decreasedby 41.79% ( P < 0.01). AT1-R silencing inhibited the proliferation of HEC-1A cells treated with 17β-E2. HEC-1A cells treat-ed with 10 -8mol/L 17β-E2 for 10 min had a decreased number of cells in G1 phase and an increased number of cells in Sphase ( P < 0.05). AT1-R silencing reversed the promoting effect of 17β-E2 on the cell cycle, increasing the number of cellsin G1 phase and decreasing the number of cells in S phase ( P < 0.05). After treatment with 10 -8mol/L 17β-E2 for 10 min,the expression of ERK1/2 protein in the HEC-1A cell line was increased and AT1-R silencing decreased the expression ofERK1/2 protein. Conclusion: AT1-R plays an important role in 17β-E2-induced proliferation and cell cycle progression in en-dometrial carcinoma cell line HEC-1A, which may be related to decreased expression of ERK1/2.Keywords Angiotensin receptor 1; 17β-estrogen; ERK1/2; HEC-1A