雌激素受体ERα36基因敲低影响PC12细胞分化相关蛋白的表达  

Knock-down of ERα36 impacts the expression of differentiation protein in PC12 cells

作　　者：马依妮[1] 韩丹女[1] 徐祎慧[1] 韩朝[1,2] 梁小峰[1] 刘晶[2] 嵇志红[3] 邹萍[4] 王兆一 邹伟[1,6] 

机构地区：[1]辽宁师范大学生命科学学院,大连116081 [2]大连医科大学附属第一医院中英再生医学中心,大连116081 [3]大连大学医学院,大连116622 [4]大连大学师范学院,大连116622 [5]克瑞顿大学癌症中心,美国内布拉斯加州68178 [6]辽宁师范大学发展和教育心理学研究中心,大连116029

出　　处：《生理学报》2012年第3期282-288,共7页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China (No. 30970353);International Science and Technology Cooperation Projects of Ministry of Science and Technology;China (No. 2010DFR30850)

摘　　要：ERα36作为一种新近鉴定出的雌激素受体亚型,广泛表达在多种组织、细胞中,参与肿瘤细胞的生长、增殖、分化过程,但其在神经系统中的表达及其功能尚不明确。本文以PC12高分化(PC12D)细胞和PC12未分化(PC12unD)细胞为研究对象,利用ERα36-shRNA质粒转染细胞建立ERα36敲低细胞模型(PC12D-36L, PC12unD-36L),通过免疫细胞荧光法、Westernblot观察神经干细胞特征标志蛋白Nestin、β-微管蛋白Ⅲ (β-tubulinIII)和神经特异性核蛋白Neu-N的表达变化。结果显示,PC12D和PC12unD细胞内均有ERα36的表达;与PC12D细胞相比,PC12unD细胞中Nestin表达较高,β-tubulinIII表达较低。敲低ERα36可降低PC12unD细胞的Nestin的表达水平,而升高β-tubulinIII和Neu-N的表达水平;敲低ERα36对PC12D细胞的上述三种蛋白的调节作用则相反。以上结果表明,敲低ERα36可抑制高分化细胞分化,促使未分化细胞分化,提示ERα36可能参与神经细胞分化的双重调控作用。ERα36 is a novel subtype of estrogen receptor alpha（ERα） known to play an important role in breast cancer development and widely expressed in normal tissues and cells including nerve cells.However,the expression and function of ERα36 in nerve cells have not been well elucidated.To examine whether ERα36 is involved in differentiation of nerve cells,the differentiated and undifferentiated PC12（PC12D and PC12unD） cells were used.Transfection of ERα36-shRNA plasmid into PC12 cells was performed to establish the ERα36 gene knock-down cells model.Immunocytofluorescence and Western blot were used to analyze the expression of Nestin,β-tubulinIII and Neu-N in the PC12 cells.The results showed that ERα36 was expressed in both cell types.Compared with PC12D cells,PC12unD cells showed higher expression of Nestin and lower expression of β-tubulinIII.ERα36-shRNA-mediated knock-down of ERα36 expression enhanced the expression of β-tubulinIII and Neu-N,but attenuated Nestin expressions in PC12unD cells;ERα36 knock-down in PC12D cells mediated Nestin,β-tubulinIII and Neu-N in a contrary manner.These results indicate thatERα36 knock-down appear to be associated with inhibiting differentiation in differentiated cells and promoting differentiation in undifferentiated cells,suggesting that ERα36 is a dual regulator in nerve differentiation.

关 键 词：ERα36 神经细胞 分化 

分 类 号：R363[医药卫生—病理学]