CAJ | 학술논문

目的观察人工合成酪蛋白激酶2(CK2)选择性抑制剂四溴肉桂酸(TBCA),对各种前列腺癌细胞株雄激素受体(AR)反式激活、细胞增殖和活力的效应.方法 TBCA和基因特异性小干扰RNA(siRNA)应用到前列腺癌细胞株,Alamar blue法检测细胞生长,流式细胞术检测细胞周期分布,荧光素酶基因报道分析检测AR反式激活,免疫荧光法检测AR核定位以及定量聚合酶链反应(PCR)检测AR介导的基因表达.结果 TBCA呈剂量依赖性抑制细胞增殖和细胞周期停滞在G2/M期,半抑制浓度值(IC50)为25 mol/L.TBCA阻断AR核转位和基因表达(P＜0.01).2种CK2催化亚单位都参与雄激素刺激的核转位和AR介导的基因表达(P＜0.05).结论 CK2α和CK2α’亚单位都参与了AR信号转运表达,TBCA有可能成为一种有效的前列腺癌化学治疗药物.
목적 관찰인공합성락단백격매2(CK2)선택성억제제사추육계산(TBCA),대각충전렬선암세포주웅격소수체(AR)반식격활、세포증식화활력적효응.방법 TBCA화기인특이성소간우RNA(siRNA)응용도전렬선암세포주,Alamar blue법검측세포생장,류식세포술검측세포주기분포,형광소매기인보도분석검측AR반식격활,면역형광법검측AR핵정위이급정량취합매련반응(PCR)검측AR개도적기인표체.결과 TBCA정제량의뢰성억제세포증식화세포주기정체재G2/M기,반억제농도치(IC50)위25 mol/L.TBCA조단AR핵전위화기인표체(P＜0.01).2충CK2최화아단위도삼여웅격소자격적핵전위화AR개도적기인표체(P＜0.05).결론 CK2α화CK2α’아단위도삼여료AR신호전운표체,TBCA유가능성위일충유효적전렬선암화학치료약물.
Objective To investigate the role of casein kinase 2 (CK2) selective inhibitor tribromoisocyanuric acid (TBCA) in androgen receptor (AR) transactivation,cell proliferation and viability in prostate cancer cell lines,and explore a new chemotherapy for prostate cancer.Methods Prostate cancer cell lines were prepared.With TBCA treatment,Alamar-blue assay was performed to assess cell proliferation and viability in C4-2 cells and flow cytometry was performed for cell cycle analysis in C4-2 and 22Rvl cells; while immunofluorescence staining was performed to determine AR nuclear translocationin in PC-3/AR cells with R1881 pretreatment,and luciferase gene reporter assay was performed to determine AR transactivation in LNCaP cells; Reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the mRNA level of prostate specific antigen (PSA).Results With TBCA treatment,Alamar-blue reading decreased in a dose-dependent manner,and the 50％ inhibitory concentration ( IC50 ) was around 25 mol/L.Significant G2/M arrest was detected in C4-2 and 22Rvl cells.R1881-induced AR nuclear localization was reduced significantly ( P ＜ 0.01 ).R1881 -stimulated ARE-LUC reporter activity in LNCaP cells decreased with reduced level of PSA mRNA,which was an AR endogenous target( P ＜ 0.05 ).Conclusion The selective CK2 inhibitor TBCA can dramatically inhibite cell proliferation and cause a G2/M phase arrest in prostate cancer cells.