新型营养素混合物对人前列腺癌细胞系 PC-3 和 DU-145 中尿激酶纤溶酶原激活物和基质金属蛋白酶的下调及其抑制剂的上调作用。
Down-regulation of urokinase plasminogen activator and matrix metalloproteinases and up-regulation of their inhibitors by a novel nutrient mixture in human prostate cancer cell lines PC-3 and DU-145.
机构信息
Dr Rath Research Institute, Cancer Division, Santa Clara, CA, USA.
出版信息
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。
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