Dr Rath Research Institute, Cancer Division, Santa Clara, CA, USA.
Oncol Rep. 2011 Dec;26(6):1407-13. doi: 10.3892/or.2011.1434. Epub 2011 Aug 24.
Strong clinical and experimental evidence shows that elevated levels of urokinase plasminogen activators (u-PA) and matrix metalloproteinases (MMPs) are associated with prostate cancer progression, metastasis and shortened survival in patients. MMP activities are regulated by specific tissue inhibitors of metalloproteinases (TIMPs). A nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract showed anticancer activity against a number of cancer cell lines. Our main objective was to study the effect of NM on the activity of u-PA, MMPs and their inhibitor TIMPs on human prostate cancer cell lines PC-3 and DU-145. Human prostate cancer cell lines PC-3 and DU-145 (ATCC) were grown in MEM media with 10% FBS and antibiotics in 24‑well tissue culture plates. At near confluence, the cells were treated with NM at 0-1000 µg/ml in triplicate at each concentration. Analysis of u-PA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. Both PC-3 and DU-145 prostate cancer cell lines demonstrated u-PA activity (subunits 1 and 2, corresponding to 35 and 33 kDa). Prostate cancer cell line PC-3 secretion of u-PA subunit 1 was decreased by 65% at NM 500 µg/ml and subunit 2 by 100% at NM 50 µg/ml. Prostate cancer cell line DU-145 secretion of u-PA subunit 1 was decreased by 97% at NM 500 µg/ml and subunit 2 by 100% at NM 100 µg/ml. Untreated PC-3 showed two bands for MMP-2 and MMP-9. NM inhibited their expression in a dose-dependent manner. The activity of MMP-2 and MMP-9 was significantly inhibited at 250 µg/ml with total inhibition at 500 µg/ml. DU-145 cells did not exhibit MMP activity. Activity of TIMPs was up-regulated in both prostate cancer cell lines in a dose-dependent manner. Minimum activity was expressed at 50 µg/ml NM and maximum at 1000 µg/ml. Correlation analyses revealed a positive correlation between u-PA and MMPs and a negative correlation between u-PA/MMPs and TIMPs. These results suggest NM as a potential anticancer agent since it targets invasive parameters of prostate cancer.
强有力的临床和实验证据表明，尿激酶纤溶酶原激活物（u-PA）和基质金属蛋白酶（MMPs）水平升高与前列腺癌的进展、转移和患者生存时间缩短有关。MMP 的活性受特定的金属蛋白酶组织抑制剂（TIMPs）调节。一种含有赖氨酸、脯氨酸、抗坏血酸和绿茶提取物的营养混合物（NM）对多种癌细胞系表现出抗癌活性。我们的主要目标是研究 NM 对人前列腺癌细胞系 PC-3 和 DU-145 的 u-PA、MMPs 及其抑制剂 TIMPs 活性的影响。人前列腺癌细胞系 PC-3 和 DU-145（ATCC）在含 10% FBS 和抗生素的 MEM 培养基中于 24 孔组织培养板中生长。当接近汇合时，将细胞在 0-1000μg/ml 的 3 个浓度下进行 NM 处理，每个浓度重复 3 次。通过纤维蛋白溶酶谱法分析 u-PA 活性，通过明胶酶谱法分析 MMPs，通过反向酶谱法分析 TIMPs。PC-3 和 DU-145 前列腺癌细胞系均表现出 u-PA 活性（亚单位 1 和 2，分别对应于 35 和 33 kDa）。PC-3 前列腺癌细胞系 u-PA 亚单位 1 的分泌在 NM 500μg/ml 时降低了 65%，亚单位 2 在 NM 50μg/ml 时降低了 100%。DU-145 前列腺癌细胞系 u-PA 亚单位 1 的分泌在 NM 500μg/ml 时降低了 97%，亚单位 2 在 NM 100μg/ml 时降低了 100%。未经处理的 PC-3 显示出 MMP-2 和 MMP-9 的两条带。NM 以剂量依赖性方式抑制其表达。MMP-2 和 MMP-9 的活性在 250μg/ml 时显著抑制，在 500μg/ml 时完全抑制。DU-145 细胞没有表现出 MMP 活性。两种前列腺癌细胞系的 TIMPs 活性均呈剂量依赖性上调。在 NM 50μg/ml 时表达最低活性，在 NM 1000μg/ml 时表达最高活性。相关性分析显示 u-PA 与 MMPs 呈正相关，u-PA/MMPs 与 TIMPs 呈负相关。这些结果表明 NM 作为一种潜在的抗癌剂，因为它针对前列腺癌的侵袭性参数。
