摘要
目的研究表没食子儿茶素没食子酸酯(EGCG)联合比卡鲁胺诱导人前列腺癌细胞LNCaP发生凋亡及抑制其增殖的作用,并探讨其机制。方法不同浓度EGCG、比卡鲁胺及两者联合处理LNCaP细胞后,通过MTT测定细胞增殖抑制情况;流式细胞仪检测凋亡率;Western印迹技术检测雄激素受体(AR)、前列腺特异性抗原(PSA)、有活性caspase-3的表达变化。结果 EGCG(25 μmol/L、50 μmol/L、100 μmol/L)、比卡鲁胺(30 μmol/L)单独及两者联合均能显著抑制LNCaP细胞增殖,诱导其凋亡,与对照组相比P<0.01,且联合组与两者单独应用之间比较均P<0.01,说明两者联合具有协同作用。EGCG(50μmol/L)、比卡鲁胺(30 μmol/L)单独及两者联合作用于LNCaP细胞72h后,AR、PSA表达明显减少,有活性caspase-3蛋白表达明显增多。结论 EGCG、比卡鲁胺单独及两者联合可抑制LNCaP细胞增殖,诱导其凋亡,且两者联合有协同作用。EGCG和比卡鲁胺诱导LNCaP细胞凋亡可能通过AR、caspase-3介导,PSA表达减少对判断前列腺癌的治疗效果具有重要的意义。
Objective To study the effect of epigallocatcchin-3-gallate （EGCG） combined with bicalutamide on proliferation and apoptosis of human prostate cancer androgen-dependent cell lines LNCaP and its mechanism. Methods LNCaP cells were treated respectively with different concentration of EGCG, bicalutamide and their combination. The proliferation and apoptosis of the treated cells were measured by methylthiazolyl tetrazolium （MTT） assay and flow cytometry （FCM）. The expression of androgen receptor （AR）, prostate-specific antigen （PSA） and cleaved caspase-3 were deteted by Western blot. Results The proliferation of LNCaP cells was significant inhibited and the apoptosis of LNCaP cells was significant induced under treatment of EGCG （ with a concentration of 25 μmol/L, 50 μmol/L, 100 μmol/L） and bicalutamide （with a concentration of 30μmol/L） alone, as well as their combination, compared with that of controls （P〈0.01）. The proliferation and apoptosis of the treated cells in the combined treatment group were significantly different from that of other alone use group （P〈0.01）, which indicated that there was a synergistic effect of the two agents in the proliferation and apoptosis of LNCaP. The expressions of AR and PSA were apparently downregulated, as well as the expression of cleaved caspase-3 was upregulated after treatment of EGCG（50 μmol/L）,bicalutamide （30μ mol/L） alone and in combination for 72 hours. Conclusion The proliferation of LNCaP cells was significant inhibited and the apoptosis of LNCaP cells was significant induced under treatment of EGCG and bicalutamide alone, as well as their combination, and a synergistic effect of EGCG and bicalutamide was found in the proliferation and apoptosis of LNCaP. The apoptosis of LNCaP cells induced by EGCG and bicalutamide might be mediated by AR and caspase-3. The level of PSA is useful to monitor the state of patient＇s tumor burden.
出处
《中国男科学杂志》
CAS
CSCD
2010年第6期29-32,35,共5页
Chinese Journal of Andrology
