Bemis Debra L, Capodice Jillian L, Anastasiadis Aristotelis G, Katz Aaron E, Buttyan Ralph
Department of Urology, Columbia University Medical Center, New York, NY 10032, USA.
Nutr Cancer. 2005;52(2):202-12. doi: 10.1207/s15327914nc5202_10.
Cyclooxygenase (COX) inhibitors have suppressive effects on several types of cancer cells including prostate cancer. In this study, we considered the potential COX-inhibitory activity of a unique anti-inflammatory herbal preparation (Zyflamend; New Chapter, Inc., Brattleboro, VT) and analyzed its effects on the human prostate cancer cell line LNCaP. COX inhibitory activity of Zyflamend was determined by a spectrophotometric-based assay using purified ovine COX-1 and COX-2 enzymes. Effects of Zyflamend on LNCaP cell growth and apoptosis in vitro were assessed by cell counting, Western blot detection of poly ADP-ribose polymerase (PARP) cleavage, and measurement of caspase-3 activity in treated and control cell extracts. Western blotting techniques were conducted to determine the effects of this herbal preparation on the expression of the cell signaling proteins, p21, androgen receptor (AR), phospho-protein kinase C (pPKC)(alpha/beta), and phospho (p)Stat3. The phospohorylation status of several signal transduction phosphoproteins was profiled using a high-throughput phosphoprotein screening assay in treated cells and compared to controls. Zyflamend dramatically decreased COX-1 and COX-2 enzymatic activity. Elevated p21 expression coincided with attenuated cell growth following treatment of LNCaP cells with Zyflamend. PARP cleavage fragments were evident, and caspase-3 activity was upregulated over the control indicating the ability of Zyflamend to induce apoptosis of these cells. Androgen receptor expression levels declined by 40%, and decreases were observed in the active forms of Stat3 and PKC(alpha/beta) in Zyflamend-treated LNCaP cells. Zyflamend inhibited both COX-1 and COX-2 enzymatic activities, suppressed cell growth, and induced apoptosis in LNCaP cells. However, our data suggests that the effects are likely due to COX-independent mechanisms potentially involving enhanced expression of p21 and reduced expression of AR, pStat3, and pPKC(alpha/beta).
环氧化酶（COX）抑制剂对包括前列腺癌在内的多种癌细胞具有抑制作用。在本研究中，我们考察了一种独特的抗炎草药制剂（Zyflamend；新章公司，佛蒙特州布拉特尔伯勒）潜在的COX抑制活性，并分析了其对人前列腺癌细胞系LNCaP的影响。使用纯化的绵羊COX - 1和COX - 2酶，通过基于分光光度法的测定来确定Zyflamend的COX抑制活性。通过细胞计数、蛋白质免疫印迹法检测聚ADP - 核糖聚合酶（PARP）裂解以及测量处理组和对照组细胞提取物中的半胱天冬酶 - 3活性，评估Zyflamend对LNCaP细胞体外生长和凋亡的影响。采用蛋白质免疫印迹技术来确定这种草药制剂对细胞信号蛋白p21、雄激素受体（AR）、磷酸化蛋白激酶C（pPKC）（α/β）和磷酸化（p）Stat3表达的影响。使用高通量磷蛋白筛选测定法分析处理细胞中几种信号转导磷蛋白的磷酸化状态，并与对照组进行比较。Zyflamend显著降低了COX - 1和COX - 2的酶活性。用Zyflamend处理LNCaP细胞后，p21表达升高与细胞生长减弱同时出现。PARP裂解片段明显，且半胱天冬酶 - 3活性相对于对照组上调，表明Zyflamend能够诱导这些细胞凋亡。雄激素受体表达水平下降了40%，并且在Zyflamend处理的LNCaP细胞中，观察到Stat3和PKC（α/β）的活性形式减少。Zyflamend抑制了COX - 1和COX - 2的酶活性，抑制了LNCaP细胞的生长并诱导其凋亡。然而，我们的数据表明，这些作用可能归因于不依赖COX的机制，可能涉及p21表达增强以及AR、pStat3和pPKC（α/β）表达降低。
