天津医药
天津醫藥
천진의약
Tianjin Medical Journal
2015年
11期
1253-1257
,共5页
刁小伟%李园%皮玉瑞%李同辉%刘平%卢山
刁小偉%李園%皮玉瑞%李同輝%劉平%盧山
조소위%리완%피옥서%리동휘%류평%로산
促红细胞生成素产生的肝细胞A型受体3%双氢睾酮%受体,雄激素%前列腺肿瘤%调节
促紅細胞生成素產生的肝細胞A型受體3%雙氫睪酮%受體,雄激素%前列腺腫瘤%調節
촉홍세포생성소산생적간세포A형수체3%쌍경고동%수체,웅격소%전렬선종류%조절
EphA3%dihydrotestosterone%receptors,androgen%prostatic neoplasms%regulation
目的：探讨雄激素依赖性前列腺癌细胞中促红细胞生成素产生肝细胞A型受体（EphA）3表达和雄激素受体（AR）信号通路的关联。方法首先通过逆转录聚合酶链式反应（RT-PCR）和蛋白免疫印迹实验（Western blot）分别检测前列腺癌细胞LNCaP和22Rv1中EphA3和AR的mRNA及蛋白水平，然后用双氢睾酮（DHT）刺激48 h，测定细胞EphA3、AR和前列腺特异抗原（PSA）表达水平的变化。同时，将成功构建的荧光素酶报告基因重组质粒EphA3-Luc（-789~+146）与AR表达质粒pcDNA3.1（+）-AR或AR特异性小分子干扰RNA（siAR）共转染22Rv1细胞，分析AR对EphA3转录活动的影响。结果 EphA3在前列腺基质细胞WPMY-1中表达水平极低，但在前列腺癌细胞LNCaP和22Rv1中则明显升高，与AR表达模式相似。10 nmol/L DHT不仅明显上调22Rv1细胞中AR、PSA和EphA3表达水平，也显著升高LNCaP细胞中相关基因和蛋白水平。而且细胞内不同AR表达水平会显著影响EphA3启动子活性。结论 AR通过提高EphA3启动子活性上调EphA3表达水平。
目的：探討雄激素依賴性前列腺癌細胞中促紅細胞生成素產生肝細胞A型受體（EphA）3錶達和雄激素受體（AR）信號通路的關聯。方法首先通過逆轉錄聚閤酶鏈式反應（RT-PCR）和蛋白免疫印跡實驗（Western blot）分彆檢測前列腺癌細胞LNCaP和22Rv1中EphA3和AR的mRNA及蛋白水平，然後用雙氫睪酮（DHT）刺激48 h，測定細胞EphA3、AR和前列腺特異抗原（PSA）錶達水平的變化。同時，將成功構建的熒光素酶報告基因重組質粒EphA3-Luc（-789~+146）與AR錶達質粒pcDNA3.1（+）-AR或AR特異性小分子榦擾RNA（siAR）共轉染22Rv1細胞，分析AR對EphA3轉錄活動的影響。結果 EphA3在前列腺基質細胞WPMY-1中錶達水平極低，但在前列腺癌細胞LNCaP和22Rv1中則明顯升高，與AR錶達模式相似。10 nmol/L DHT不僅明顯上調22Rv1細胞中AR、PSA和EphA3錶達水平，也顯著升高LNCaP細胞中相關基因和蛋白水平。而且細胞內不同AR錶達水平會顯著影響EphA3啟動子活性。結論 AR通過提高EphA3啟動子活性上調EphA3錶達水平。
목적：탐토웅격소의뢰성전렬선암세포중촉홍세포생성소산생간세포A형수체（EphA）3표체화웅격소수체（AR）신호통로적관련。방법수선통과역전록취합매련식반응（RT-PCR）화단백면역인적실험（Western blot）분별검측전렬선암세포LNCaP화22Rv1중EphA3화AR적mRNA급단백수평，연후용쌍경고동（DHT）자격48 h，측정세포EphA3、AR화전렬선특이항원（PSA）표체수평적변화。동시，장성공구건적형광소매보고기인중조질립EphA3-Luc（-789~+146）여AR표체질립pcDNA3.1（+）-AR혹AR특이성소분자간우RNA（siAR）공전염22Rv1세포，분석AR대EphA3전록활동적영향。결과 EphA3재전렬선기질세포WPMY-1중표체수평겁저，단재전렬선암세포LNCaP화22Rv1중칙명현승고，여AR표체모식상사。10 nmol/L DHT불부명현상조22Rv1세포중AR、PSA화EphA3표체수평，야현저승고LNCaP세포중상관기인화단백수평。이차세포내불동AR표체수평회현저영향EphA3계동자활성。결론 AR통과제고EphA3계동자활성상조EphA3표체수평。
Objective To evaluate the relationship between liver cell type A receptor (EphA) expression and androgen receptor (AR) signaling in androgen-dependent prostate cancer cells. Methods RT-PCR and Western blot assay were used to determine mRNA and protein levels of EphA3 and AR in prostate cancer LNCaP and 22Rv1 cells, respectively. The variations of EphA3, AR and prostate specific antigen (PSA) expressions were also measured in these cells after dihydrotes?tosterone (DHT) treatment for 48 h. The constructed EphA3-Luc (-789-+146) luciferase reporter plasmid was co-transfect?ed with pcDNA3.1(+)-AR or siAR in 22Rv1 cells to analyze the effects of different AR expression levels on EphA3 tran?scription activity. Results The expression pattern of EphA3 was similar to AR, showing a lower level in prostate stromal cell line WPMY-1 and a higher level in prostate cancer cell lines LNCaP and 22Rv1. When stimulated with 10 nmol/L DHT, the expression levels of AR, PSA and EphA3 were significantly increased in 22Rv1 cells, and the protein levels of these genes were also increased in LNCaP cells. Moreover, AR expression levels markedly influenced the activity of EphA 3 pro?moter. Conclusion AR up-regulates EphA3 expression by increasing the activity of EphA3 promoter.
