论文部分内容阅读
目的探讨苯甲酸雌二醇对离体子宫内膜腺上皮细胞中热休克蛋白(HSP)70、90表达的影响。方法采用酶消化法,行原代子宫内膜腺上皮细胞培养。将培养的子宫内膜腺上皮细胞分为对照组(加入正常培养液)、苯甲酸雌二醇组、苯甲酸雌二醇+雌激素拮抗剂ICI182780组和ICI182780组,各组细胞分别作用6、12、18和24h。四甲基偶氮唑蓝还原法测定不同浓度的苯甲酸雌二醇及ICI182780对离体子宫内膜腺上皮细胞生长的影响,免疫组化法及图像分析法测定苯甲酸雌二醇和ICI182780对腺上皮细胞中HSP70、90表达的影响。结果苯甲酸雌二醇可刺激腺上皮细胞生长,细胞增殖率随着作用时间的延长和药物浓度升高而升高,浓度为10-9、10-8、10-7、10-6mol/L的苯甲酸雌二醇作用24h时,平均细胞增殖率分别为(170±9)%、(207±11)%、(231±12)%、(257±10)%,均明显高于上述各浓度苯甲酸雌二醇作用6h时的细胞增殖率[(117±13)%、(129±10)%、(146±10)%、(176±6)%],差异有统计学意义(P<0.05);浓度为10-9、10-8、10-7、10-6mol/L的苯甲酸雌二醇作用24h时,苯甲酸雌二醇+ICI182780组与苯甲酸雌二醇组比较,细胞平均增殖率均明显降低,分别为(137±4)%、(145±10)%、(151±9)%、(167±3)%,差异有统计学意义(P<0.05)。苯甲酸雌二醇可刺激细胞中HSP90的表达,这种刺激作用同样存在药物浓度和作用时间依赖性,苯甲酸雌二醇浓度为10-9、10-8、10-7、10-6mol/L,作用24h时HSP90表达的相对不透光度分别为(64.8±10.7)%、(75.9±12.6)%、(80.4±8.2)%、(83.2±7.6)%,明显高于相同浓度、相同作用时间时对照组的(28.0±3.3)%、(29.0±5.6)%、(29.0±5.0)%和(30.0±6.4)%,差异也有统计学意义(P<0.05);浓度为10-9、10-8、10-7、10-6mol/L的苯甲酸雌二醇作用24h时,苯甲酸雌二醇+ICI182780组与苯甲酸雌二醇组比较,腺上皮细胞中HSP90表达明显降低,分别为(28.2±2.1)%、(29.7±3.2)%、(35.0±4.7)%、(34.7±6.5)%。ICI182780不影响雌二醇对腺上皮细胞中HSP70表达的抑制作用。结论苯甲酸雌二醇对子宫内膜腺上皮细胞增殖及HSP90表达的影响是雌激素受体依赖性的,HSP90表达可能有助于苯甲酸雌二醇刺激腺上皮细胞增殖;苯甲酸雌二醇对HSP70表达的抑制作用是非雌激素受体依赖性的,HSP70表达可能对腺上皮细胞的损伤有保护作用。
Objective To investigate the effect of estradiol benzoate on the expression of heat shock protein (HSP) 70,90 in isolated endometrial glandular epithelial cells. Methods Enzyme digestion was used to culture primary endometrial glandular epithelial cells. The cultured endometrial glandular epithelial cells were divided into control group (normal culture medium), estradiol benzoate group, estradiol benzoate + estrogen antagonist ICI182780 group and ICI182780 group, 12, 18 and 24h. The effects of different concentrations of estradiol benzoate and ICI182780 on the growth of isolated endometrial glandular epithelial cells were determined by MTT assay. The expressions of estradiol benzoate and ICI182780 were measured by immunohistochemistry and image analysis Effect of HSP70,90 expression in epithelial cells. Results Estradiol benzoate stimulated the growth of glandular epithelial cells. The cell proliferation rate increased with the prolongation of action time and the increase of drug concentration. The concentrations of 10-9,10-8,10-7,10-6 mol / L (207 ± 11)%, (231 ± 12)%, (257 ± 10)%, respectively, were significantly higher than those of the above (P <0.01), (129 ± 10)%, (146 ± 10)%, (176 ± 6)%] at 6 h after treatment with estradiol benzoate. The difference was statistically significant (P <0.05). Compared with the estradiol benzoate group, the estradiol benzoate + ICI182780 group at the concentration of 10-9,10-8,10-7,10-6mol / L estradiol benzoate for 24h, (137 ± 4)%, (145 ± 10)%, (151 ± 9)% and (167 ± 3)%, respectively. The difference was statistically significant (P <0.05). Estradiol benzoate can stimulate the expression of HSP90 in cells, this stimulus also exists drug concentration and time-dependent effects, estradiol benzoate concentration of 10-9,10-8,10-7,10-6 mol / L, the relative opacity of HSP90 expression was (64.8 ± 10.7)%, (75.9 ± 12.6)%, (80.4 ± 8.2)% and (83.2 ± 7.6)% respectively at 24h and was significantly higher than that of the same concentration (28.0 ± 3.3)%, (29.0 ± 5.6)%, (29.0 ± 5.0)% and (30.0 ± 6.4)% in the control group at the time of action, the difference was also statistically significant (P <0.05) , 10-8,10-7,10-6mol / L estradiol benzoate 24h, estradiol benzoate + ICI182780 group compared with estradiol benzoate group, HSP90 expression in glandular epithelial cells was significantly decreased, (28.2 ± 2.1)%, (29.7 ± 3.2)%, (35.0 ± 4.7)% and (34.7 ± 6.5)%, respectively. ICI182780 does not affect the inhibitory effect of estradiol on HSP70 expression in glandular epithelial cells. Conclusion The effect of estradiol benzoate on the proliferation of endometrial glandular epithelial cells and the expression of HSP90 is estrogen receptor-dependent. The expression of HSP90 may contribute to the stimulation of glandular epithelial cells by estradiol benzoate. Estradiol benzoate The inhibition of HSP70 expression is non-estrogen receptor-dependent, HSP70 expression may have a protective effect on the damage of glandular epithelial cells.