雌激素受体拮抗剂对垂体腺瘤GH3细胞增殖、凋亡、催乳素分泌及雌激素受体蛋白表达的影响  

Differential effects of estrogen receptor antagonists on pituitary GH3 cell proliferation, apoptosis, PRL secretion, and ER protein expression

作　　者：王婉[1] 赵兴利[2] 刘晶[2] 韦博[2] 郭永川[2] 

机构地区：[1]吉林大学白求恩医学院免疫学系,长春130021 [2]吉林大学中日联谊医院神经外科

出　　处：《中华神经外科杂志》2012年第3期285-290,共6页Chinese Journal of Neurosurgery

基　　金：基金项目：国家自然科学基金青年科学基金项目（30801176）;吉林省自然科学基金面上项目（201115096）

摘　　要：目的通过检测雌激素受体拮抗剂对垂体腺瘤GH3细胞增殖、凋亡、催乳素（PRL）分泌及雌激素受体蛋白表达的影响，探讨雌激素在垂体催乳素腺瘤发生、发展中的作用机制。方法对去激素培养条件及加入外源性E2培养条件下的GH3细胞采用不同方法进行检测：应用MTT法检测OHTam与ICI对GH3细胞增殖的影响；ELISA法检测对GH3细胞PRL分泌的影响；流式细胞仪测定细胞凋亡率；Westernblot检测对GH3细胞ERd、ERB表达的影响。结果E2对GH3细胞增殖有显著的生长刺激作用，低浓度E2（10^-12mol／L）即显著高于去激素培养组；OHTam与ICI对GH3细胞增殖有抑制作用，当给药浓度达到10^-6mol／L时，增殖指数分别为0．62和0．47，呈剂量依赖性；OHTam与ICI可抑制E2对GH3细胞的生长刺激作用；以上结果与对照组相比差异有统计学意义（P〈0．05）。E2可促进GH3细胞的PRL的分泌，随着E2浓度的增高，PRL的分泌增加；OHTam与ICI可抑制GH3细胞PRL的分泌，并且能够抑制E2的促进作用。OHTam与ICI能够诱导GH3细胞凋亡，以早期凋亡为主。GH3细胞有ER仪与ERβ的表达，E2可上调ERp的表达水平，ICI可下调ER仅的表达水平，而OHTam对ER仪与ERB的表达水平无显著影响。结论雌激素受体拮抗剂是通过抑制细胞增殖、PRL分泌及抗雌激素而发挥抑瘤作用，而ICI的作用还与下调ERα的表达有关。Objective To detect the effects of estrogen receptor antagonists on pituitary GH3 cell proliferation, apoptosis, prolactin （PRL） secretion, and estrogen receptor （ ER ） isoforms expressions, and to investigate the mechanisms of exogenous estrogen（ 17β- estradiol, E2） in prolactinoma tumorigenesis and development. Method Different methods had been used to test the GH3 ceils cultured with hormone - deprived＆phenol red - free medium or exogenous E2 - added medium ： MTF assays were used to detect the inhibitory effects of OHTam and ICI on proliferation of GH3 cells;rat PRL ELISA kit was applied to the detection of PRL secretion ; flow cytometry was used to detect OHTam and ICI induced cell apoptosis ; Western blot analysis was applied to the detection of ERct and ERβ expressed in GH3 cells. Results E2 had significant stimulatory effect on growth of GH3 cells, and the effect worked even at low concentration of 10^- 12 mol/L compared to the hormone- deprived medium cultured GH3 cells. OHTam and ICI could inhibit GH3 cell proliferation as dose -dependent manner, when the concentration rising to 10-6 mol/L, the cell proliferation index were 0. 62 and 0. 47 respectively;OHTam and ICI could suppressed the stimulatory effect of E2 on GH3 cell proliferation. All the results had statistical significance （ P 〈 0. 05 ）. E2 could increase PRL secretion in GH3 cells and the effect appeared to be dose -dependent;OHTam and ICI decreased PRLsecretion, and could inhibit the promotion effect of E2. OHTam and ICI could induced GH3 cells apoptosis, mostly early apoptosis. The expression of ERα and ERβ in GH3 cells were observed, E2 up - regulated ER~ expression level, and ICI obviously down- regulated the expression of ERα, however, OHTam couldn＇t regulate the expression levels of ERα and ERβ. Conclusion Estrogen receptor antagonists can inhibit GH3 cell proliferation and PRL secretion, induce cell apoptosis, and inhibit the effect of E2, and thus, generate antitumor effects. In addition, ICI can down - re

关 键 词：催乳素腺瘤 雌激素类 催乳素 受体 雌激素 雌激素受体拮抗剂 

分 类 号：R736.4[医药卫生—肿瘤]