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目的研究雌二醇(E2)影响小鼠结肠腺癌细胞系MCA-38细胞增殖与凋亡及雌激素受体(ER)表达变化的特点与机制。方法将MCA-38细胞分为5个实验组(分别含E2浓度为0.01 nmol/L、0.1 nmol/L、1 nmol/L、10 nmol/L、100 nmol/L的培养液培养)和空白对照组,体外培养24 h。检测各组肿瘤细胞E2处理前后ERα和ERβ表达(Western blot法)、细胞增殖(MTT法)以及caspase-3表达(Western blot法)变化。结果 MCA-38细胞表达ERα、ERβ,后者是其主要ER。不同剂量的E2均影响ERα和ERβ表达,E2上调ERα蛋白表达,0.01 nmol/L、0.1 nmol/L、1 nmol/L浓度范围的E2作用最明显,与对照组相比分别使ERα蛋白表达量增加了4.7%、5.5%和5.9%(P<0.05)。E2则使ERβ表达下降,0.1 nmol/L、1 nmol/L组最明显,分别使ERβ蛋白表达量下降了10.2%和3.9%(P<0.05);与对照组相比,0.01 nmol/L、0.1 nmol/L、1 nmol/L、10 nmol/L浓度E2使MCA-38细胞数目分别增加了20.47%、25.29%、37.59%和30.95%(P<0.05);经E2处理,MCA-38细胞的caspase-3蛋白表达量明显下降,其中以0.1 nmol/L和1 nmol/L组最为明显,分别降低20.2%和32.9%(P<0.05)。结论 E2对MCA-38细胞的ERα和ERβ表达、细胞增殖及凋亡均产生影响,MCA-38细胞增殖和凋亡分别与ERα和ERβ表达变化密切相关。
Objective To investigate the characteristics and mechanisms of estradiol (E2) on proliferation, apoptosis and estrogen receptor (ER) expression in mouse colon adenocarcinoma cell line MCA-38. Methods MCA-38 cells were divided into five experimental groups (culture medium containing E2 concentration of 0.01 nmol / L, 0.1 nmol / L, 1 nmol / L, 10 nmol / L and 100 nmol / L) and blank control Group, cultured in vitro 24 h. The expressions of ERα and ERβ in the tumor cells before and after treatment were detected by Western blot, MTT and caspase-3 expression. Results MCA-38 cells expressed ERα, ERβ, the latter being their major ER. E2 at different doses affected the expression of ERα and ERβ, and E2 up-regulated the expression of ERα protein. E2 at concentrations of 0.01 nmol / L, 0.1 nmol / L and 1 nmol / L had the most obvious effect. Compared with the control group, ERα protein expression An increase of 4.7%, 5.5% and 5.9% (P <0.05). E2 decreased the expression of ERβ. The expression of ERβ in 0.1 nmol / L and 1 nmol / L groups decreased by 10.2% and 3.9%, respectively (P <0.05). Compared with the control group, the expression of ERβ decreased by 0.01 nmol / L, The number of MCA-38 cells were increased by 20.47%, 25.29%, 37.59% and 30.95% (P <0.05) by 0.1 nmol / L, 1 nmol / L and 10 nmol / (P <0.05). The expression of caspase-3 protein was significantly decreased in the group of 0.1 nmol / L and 1 nmol / L, which was decreased by 20.2% and 32.9%, respectively. Conclusion E2 can affect the expression of ERα and ERβ, proliferation and apoptosis of MCA-38 cells. The proliferation and apoptosis of MCA-38 cells are closely related to the changes of ERα and ERβ.