雌激素受体α和雌激素受体β在调控青少年椎体生长板软骨细胞分化和增殖中的相互作用

The interaction of ERα and ERβ in the regulation of differentiation and proliferation of vertebral growth plate chondrocyte

目的初步探索雌激素受体(ER)α和ERβ基因在青春期椎体生长板软骨细胞分化和增殖中相互作用,以及Smad4基因在ERα和ERβ调控软骨细胞Runx2和BMP2基因表达过程中的信号传递作用。方法设计ERα、ERβ和Smad4mRNA打靶的RNAi慢病毒载体并包装成为慢病毒颗粒。以小鼠椎体生长板软骨细胞为对象,采用分层设计的方法进行分组,分别转染目的基因RNAi载体,研究Smad4基因在椎板软骨细胞中介导ERα和ERβ调控Runx2和BMP2基因的表达的作用。分析各组细胞生长曲线;检测各组细胞Runx2、BMP2表达的变化。结果于转染后第3、5、7、9天时,各组细胞吸光度值(A490)在多个组间比较差异有统计学意义(P<0.05)。各组间Runx2基因表达相对于空白对照组改变的倍数由高到低的顺序与细胞生长曲线基本相同。而BMP2基因表达由高到低的顺序与Runx2不同,多个亚组间差异有统计学意义(P<0.05)。然而,无论ERαRNAi组还是ERβRNAi组中,Smad4RNAi亚组与Smad4正常亚组比较,BMP2基因表达均差异无统计学意义(P>0.05)。结论 ERα和ERβ均具有调控椎体生长板软骨细胞分化与增殖的作用,且ERα的调控作用大于ERβ。在ERα和ERβ均正常表达时,ERβ具有部分抑制ERα的功能;当ERα表达显著降低时,ERβ可以部分补偿ERα的功能。另外,ERβ通过Smad4信号传递增强软骨细胞Runx2基因表达,但还可能存在非Smad4信号传递通路。然而。

Objective To explore the interaction of the ERαand ERβin regulating mice adolescence verte‐bral epiphyseal plate chondrocyte differentiation and proliferation ,and the role of smad4 signaling transduction in ERαand ERβregulating the Runx2 and BMP2 expression.Methods The siRNA sequences targeting and silencing ERα,ERβand Smad4 mRNA were designed which were constructed into gene silencing lentiviral vector and pack‐aged as lentiviral particles .Mice vertebral growth plate chondrocytes are extracted ,grouped and were grouped with hierarchical design and were transfected target gene RNAi vectors to observe the smad4 signaling in ERαand ERβregulating Runx2 and BMP2 gene expression in chondrocytes .The cells growth curves were plotted and the Runx2 and BMP2 expression was detected in each group .Results The A value in 10 subgroups at 3th ,5th ,7th and 9th day after transfected were significantly different among groups （ P 〈 0 .05） .Descending order of the Runx2 gene expression in each subgroup was basically the same as A value .The descending order of BMP2 gene expression was not same as Runx2 .Whether in ERβRNAi group or in ERαRNAi group .There were no significant differences between Smad4RNAi subgroup and Smad4 normal subgroup（ P 〉0 .05）.Conclusion Both ERα and ERβcould regulate vertebral growth plate cartilage cell differentiation and proliferation .The role of regulation of ERαwas greater than ERβ.When the expression of ERα and ERβ were normal ,ERβ has partially inhibit ERαfunction .However ,when the expression of ERα significantly reduced ,ERβ can partially compensate for ERαfunction .ERβalso enhanced the chondrocyte differentiation and proliferation by partly reinforced Runx2 gene ex‐pression by Smad4 signaling ,but it may also by non‐Smad4 signaling pathway .However ,the regulation of ERαand ERβon BMP2 gene expression was not through Smad4 signaling pathway.