子宫内膜癌细胞中细胞膜雌激素受体表达的初步研究

目的探讨子宫内膜癌细胞中细胞膜雌激素受体的表达情况。方法对子宫内膜癌高分化细胞系Ishikawa和中分化细胞系HEC-1A的细胞膜和整个细胞(包括细胞膜、细胞质和细胞核)雌激素受体(ER)α、β表达情况进行研究。用来检测细胞膜雌激素受体的细胞经固定处理,用来检测整个细胞ER的细胞经渗透处理,分别对固定与渗透处理的Ishikawa和HEC-1A细胞进行间接免疫荧光染色,并在荧光显微镜下观察ER的分布。另对Ishikawa和HEC-1A活细胞(用来检测细胞膜雌激素受体)、渗透处理的Ishikawa和HEC-1A细胞(用来检测整个细胞ER)进行间接免疫荧光染色,并用流式细胞仪检测ER的相对荧光强度来表示ER的表达强度。以上实验均以未用一抗的细胞为阴性对照。结果荧光显微镜下观察显示,固定处理的Ishikawa和HEC-1A细胞膜均可见ERα和ERβ标记的荧光,渗透处理的Ishikawa和HEC-1A细胞,除细胞膜外,在细胞内部也可见ERα和ERβ标记的荧光。流式细胞仪检测显示,Ishikawa活细胞ERα及ERβ相对荧光强度分别为1．09±0．21和1．27±0．33,高于阴性对照(均为1．00),但两者分别与阴性对照比较,差异均无统计学意义(P＞0．05);渗透处理的Ishikawa细胞ERα及ERβ相对荧光强度分别为4．21±0．34和4．69±1．96,均高于活细胞,两者分别与活细胞比较,差异均有统计学意义(P＜0．05)。HEC-1A活细胞ERα及ERβ相对荧光强度分别为1．58±0．13和1．49±0．04,高于阴性对照(均为1．00),与阴性对照分别比较,差异均有统计学意义(P＜0．05);渗透处理的HEC-1A细胞ERα及ERβ相对荧光强度分别为2．34±0．33和2．52±0．15,均高于活细胞,两者分别与活细胞比较,差异均有统计学意义(P＜0．05)。结论子宫内膜癌细胞系Ishikawa和HEC-1A细胞中均存在细胞膜雌激素受体。

Preliminary study of membrane estrogen receptor expression in endometrial carcinoma cells

OBJECTIVE: To analyze if there are membrane estrogen receptors (ER) in endometrial carcinoma and if there is some relationship between membrane ER and nuclear ER. METHODS: The cell membrane and total cell ERalpha and ERbeta expressions of high and moderate differentiation endometrial carcinoma cells (Ishikawa and HEC-1A cells) were analyzed. Intermittent immunofluorescence dyeing and fluorescent microscopy were carried out with the cells treated with polyformaldehyde and Triton X-100. Intermittent immunofluorescence dyeing and flow cytometry were carried out with the live cells and the cells treated with Triton X-100 respectively. RESULTS: There were fluorescences on the membrane of the Ishikawa and HEC-1A cells which were treated with polyformaldehyde. When the cells were treated with Triton X-100, the fluorescences were also seen inside the cells. The fluorescence intensity of ERalpha and ERbeta in Ishikawa cell membrane (1.09 +/- 0.21, 1.27 +/- 0.33) was stronger than the control, but there were no significant differences (P > 0.05). When treated with Triton X-100, the total cell fluorescence intensity of ERalpha and ERbeta in Ishikawa cell (4.21 +/- 0.34, 4.69 +/- 1.96) was stronger than the membrane (P < 0.05). The ERalpha and ERbeta fluorescence intensity of HEC-1A cell membrane (1.58 +/- 0.13, 1.49 +/- 0.04) were stronger than the control (P < 0.05). The fluorescence intensity of ERalpha and ERbeta of the HEC-1A cell (2.34 +/- 0.33, 2.52 +/- 0.15) was stronger than the membrane also (P < 0.05). The membrane ERalpha fluorescence intensity of Ishikawa was lower than HEC-1A (P = 0.028). But the total cell ERalpha fluorescence intensity of Ishikawa was higher than HEC-1A (P = 0.002). CONCLUSIONS: There are membrane ER on endometrial carcinoma cells Ishikawa and HEC-1A. The membrane ER must have some similarity to the nuclear receptor. There is no direct correlation between the quantity of the membrane ER and nuclear ER.