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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。
Objective To investigate the expression of estrogen receptor in cell membrane in endometrial carcinoma cells. Methods The expression of estrogen receptor (ER) α and β in the cell membrane and the whole cell (including the cell membrane, cytoplasm and nucleus) of the highly differentiated cell line Ishikawa and the moderately differentiated cell line HEC-1A of endometrial carcinoma was studied. The cells used to detect the estrogen receptor of the cell membrane were fixed and used to detect the cells of the whole cell ER were infiltrated and subjected to indirect immunofluorescence staining on the immobilized and infiltrated Ishikawa and HEC-1A cells, respectively, under a fluorescence microscope Observe the distribution of ER. In addition, indirect immunofluorescence staining was performed on Ishikawa and HEC-1A living cells (for detecting cell membrane estrogen receptor), osmotic treated Ishikawa and HEC-1A cells (for detecting whole cell ER), and ER Relative fluorescence intensity to express the intensity of ER expression. The above experiments were used as primary antibody negative control. Results Fluorescence microscopy showed that both ERα and ERβ-labeled fluorophores, Ishikawa and HEC-1A cells infiltrated by Ishikawa and HEC-1A cell membranes were visible in the cells, and ERα and ERβ-labeled fluorescence . Flow cytometry showed that the relative fluorescence intensities of ERα and ERβ in Ishikawa were 1.09 ± 0.21 and 1.27 ± 0.33, respectively, which were higher than those in the negative control (all 1.00) (P> 0.05). The relative fluorescence intensities of ERα and ERβ in infiltrated Ishikawa cells were 4.21 ± 0.34 and 4.69 ± 1.96, respectively, both of which were high In living cells, the difference between them was statistically significant (P <0.05). The relative fluorescence intensities of ERα and ERβ in HEC-1A cells were 1.58 ± 0.13 and 1.49 ± 0.04, respectively, which were higher than those in the negative control (all 1.00). Compared with the negative control, the difference was (P <0.05). The relative fluorescence intensities of ERα and ERβ in infiltrating HEC-1A cells were 2.34 ± 0.33 and 2.52 ± 0.15, respectively, which were higher than that in living cells, both Respectively, compared with living cells, the difference was statistically significant (P <0.05). Conclusions Both endometrial cancer cell lines Ishikawa and HEC-1A cells present cell membrane estrogen receptors.