CAJ | 학술논문

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；Western blot检测对GH3细胞ERα、ERβ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ERα与ERβ的表达,E2可上调ERβ的表达水平,ICI可下调ERα的表达水平,而OHTam对ERα与ERβ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.
목적 통과검측자격소수체길항제대수체선류GH3세포증식、조망、최유소(PRL)분비급자격소수체단백표체적영향,탐토자격소재수체최유소선류발생、발전중적작용궤제.방법 대거격소배양조건급가입외원성E2배양조건하적GH3세포채용불동방법진행검측:응용MTT 법검측OHTam여ICI대GH3세포증식적영향；ELISA법검측대GH3세포PRL분비적영향；류식세포의측정세포조망솔；Western blot검측대GH3세포ERα、ERβ표체적영향.결과 E2대GH3세포증식유현저적생장자격작용,저농도E2(10-12mol/L)즉현저고우거격소배양조；OHTam여ICI대GH3세포증식유억제작용,당급약농도체도10 -6mol/L시,증식지수분별위0.62화0.47,정제량의뢰성；OHTam여ICI가억제E2대GH3세포적생장자격작용；이상결과여대조조상비차이유통계학의의(P＜0.05).E2가촉진GH3세포적PRL적분비,수착E2농도적증고,PRL적분비증가；OHTam여ICI가억제GH3세포PRL적분비,병차능구억제E2적촉진작용.OHTam여ICI능구유도GH3세포조망,이조기조망위주.GH3세포유ERα여ERβ적표체,E2가상조ERβ적표체수평,ICI가하조ERα적표체수평,이OHTam대ERα여ERβ적표체수평무현저영향.결론 자격소수체길항제시통과억제세포증식、PRL분비급항자격소이발휘억류작용,이ICI적작용환여하조ERα적표체유관.
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 cells cultured with hormone -deprived&phenol red - free medium or exogenous E2 - added medium:MTT 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 ERo 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 - 12mol/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 ERβ 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 - regulate the expression of ERα,and that may be related to its antitumor effects.