选择性雌激素受体调节剂对MC3T3-E1细胞生长分化的调节

Selective estrogen receptor modulators stimulates growth and differentiation in MC3T3-E1 cells:involvement of ER and Wnt/β-catenin signals

目的探讨选择性雌激素受体调节剂(selective estrogen receptor modulator,SERM)———雷洛昔芬对小鼠颅顶成骨细胞MC3T3-E1分化的调节及其机制。方法体外培养MC3T3-E1细胞,10-7mol/L的雷洛昔芬和雌激素受体拮抗剂ICI-182780刺激细胞,另设空白对照组,24、48、72 h后检测各指标。MTT法检测细胞增殖;微量酶标法检测细胞碱性磷酸酶(alkaline phosphatase,ALP)活性;实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction,qRT-PCR)检测细胞内β-catenin,雌激素受体α、β(estrogen receptorα、β,ERα、ERβ)mRNA的表达。结果 MC3T3-E1细胞分别加入10-7mol/L的雷洛昔芬和ICI-182780后,相对于对照组,雷洛昔芬处理组的促细胞增殖作用较强,在24 h增殖率最高,达(55.93±10.88)%;ALP活性增加,在48 h活性增加率最高,为(30.881±5.614)%;β-catenin、ERα和ERβmRNA的表达均增高,有统计学意义(P<0.05)。ICI-182780组的促细胞增殖率、ALP活性降低;β-catenin、ERα和ERβmRNA的表达均降低,有统计学意义(P<0.05)。结论雷洛昔芬可能通过上调β-catenin、ERα和ERβmRNA的表达促进MC3T3-E1细胞的增殖分化,而ICI-182780则下调β-catenin、ERα和ERβmRNA的表达抑制MC3T3-E1细胞的增殖和分化。

Objective To investigate the regulation and mechanism of selective estrogen receptor modulator （SERM） raloxifene on the growth and differentiation in MC3T3-E1 cells in vitro. Methods MC3T3-E1 cells cultured in vitro were treated with raloxifene （ 10 ^-7 mol/L） and estrogen receptor antagonist ICI-182780 （10^-7 mol/L）, respectively, and the MC3T3-E1 cells without treatment were set as control. The cell proliferation was observed by MTT assay, the activity of alkaline phosphatase （ALP） was measured by trace enzyme labeling method, and the expression of β-catenin, estrogen receptor （ER） α and ERβ were detected by qRT- PCR. Results As compared with those of the control group, the cell proliferation, ALP activity and mRNA expression of β-catenin, ERα and ERβ significantly increased in the raloxifene （10^-7 mol/L） group （P 〈 0. 05 ） and significantly decreased in the ICI-182780 group （P 〈 0. 05 ）. The cell proliferation rate was highest （55.93 ±10.88）% at 24 h, and the ALP activity of increase was highest （30. 881 ±5. 614）% at 48 h. Conclusion Raloxifene can promote the proliferation and differentiation of MC3T3-E1 cells through up-regulating the mRNA expression of β-catenin, ERα and ERβ, while IC1-182780 can inhibit this process through down-regulating the mRNA expression of β-catenin, ERα and ERβ.