CAJ | 학술논문

目的观察缺氧条件下人前列腺间质细胞体外培养细胞核膜表面雌雄激素受体表达的变化. 方法人前列腺间质细胞株WPMY-1体外培养.在缺氧或有氧培养环境中分别于接种后4、8、12、24、48 h检测其核膜雌雄激素受体基因和蛋白表达.基因表达采用RT-PCR法检测,蛋白表达采用免疫组化法检测. 结果缺氧条件下接种4h前列腺间质细胞雄激素受体mRNA相对表达量为0.35±0.01,后随时间逐渐上升至48 h的1.40±0.02,显著高于同期有氧条件下的0.27±0.01(t=28.182,P＜0.001)和0.36±0.01(t=98.108,P＜0.001),而免疫组化显示缺氧条件下雄激素受体蛋白表达自12 h开始较有氧条件下显著增加,分别为(33.72±4.19)/200个细胞和(23.84±1.31)/200个细胞(t=3.902,P=0.018).缺氧条件下12h间质细胞雌激素mRNA表达为0.39±0.01,至48 h升至0.59±0.01,显著高于同期有氧条件下的0.31±0.01(t=9.483,P＝0.001)和0.46±0.13(t=27.634,P＜0.001).4h时雌激素阳性表达上皮细胞为(13.42±0.80)/200个细胞后逐渐升至48 h的(55.16±0.41)/200个细胞,显著高于同期有氧条件下的(9.68±0.63)/200个细胞(t=6.531,P=0.003)和(22.95±0.55)/200个细胞(t=81.130,P＜0.001). 结论缺氧可刺激人前列腺间质细胞核膜表面雌雄激素受体表达的上调.
목적 관찰결양조건하인전렬선간질세포체외배양세포핵막표면자웅격소수체표체적변화. 방법 인전렬선간질세포주WPMY-1체외배양.재결양혹유양배양배경중분별우접충후4、8、12、24、48 h검측기핵막자웅격소수체기인화단백표체.기인표체채용RT-PCR법검측,단백표체채용면역조화법검측. 결과 결양조건하접충4h전렬선간질세포웅격소수체mRNA상대표체량위0.35±0.01,후수시간축점상승지48 h적1.40±0.02,현저고우동기유양조건하적0.27±0.01(t=28.182,P＜0.001)화0.36±0.01(t=98.108,P＜0.001),이면역조화현시결양조건하웅격소수체단백표체자12 h개시교유양조건하현저증가,분별위(33.72±4.19)/200개세포화(23.84±1.31)/200개세포(t=3.902,P=0.018).결양조건하12h간질세포자격소mRNA표체위0.39±0.01,지48 h승지0.59±0.01,현저고우동기유양조건하적0.31±0.01(t=9.483,P＝0.001)화0.46±0.13(t=27.634,P＜0.001).4h시자격소양성표체상피세포위(13.42±0.80)/200개세포후축점승지48 h적(55.16±0.41)/200개세포,현저고우동기유양조건하적(9.68±0.63)/200개세포(t=6.531,P=0.003)화(22.95±0.55)/200개세포(t=81.130,P＜0.001). 결론 결양가자격인전렬선간질세포핵막표면자웅격소수체표체적상조.
Objective To observe the different expressions of androgen and estrogen receptor in nucleus membrane of human prostate stromal cells under anoxic or normoxic conditions.Methods Human prostate stromal cell line WPMY-1 was cultured in vitro.At 4,8,12,24,48 h after cellswere seeded,the mRNA and protein expression of androgen receptor (AR) and estrogen receptor (ER) in prostate stromal cells were tested by RT-PCR and immunohistochemistry method,respectively.Results The exprcssions of AR and ER were significantly increased in prostate stromal cells under anoxic conditions compared with under normoxic conditions.The relative expression of AR mRNA was 0.35±0.01 in anoxic prostate stromal cells at 4 h,and increased to 1.40±0.02 at 48 h,which was higher than in normoxic prostate epithelial cells [0.27 ± 0.01 and0.36± 0.01] synchronously (t=28.182,both P ＜ 0.001).Immunohistochemistry showed significantly increased AR-positive cells under anoxic conditions as compared with under-normoxiccondition from 12 h synchronously [(33.72±4.19) per 200 cells,(23.84±1.31) per 200 cells,t=3.902,P=0.018].The expression of ER mRNA was 0.39±0.01 in prostate stromal cells at 4 h,and increased to 0.59±0.01 at 48 h under anoxic conditions,which were higher than under normoxic conditions (0.31±0.01 and 0.46±0.13) synchronously (both P＜0.001).Immunohistochemistry showed the significantly increased ER-positive cells under anoxic conditions as compared with under normoxic condition from 4 h to 48 h synchronously [(13.42±0.80) per 200 cells and (55.16±0.41) per 200 cells； (9.68±0.63) per 200 cells and (22.95±0.55) per 200 cells,t=81.130,P＜0.001].Conclusions The expression of androgen and estrogen receptors is upregulated in human prostatestromal cells under anoxic conditions.