Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050, USA.
Int J Oncol. 2014 Mar;44(3):986-92. doi: 10.3892/ijo.2013.2235. Epub 2013 Dec 30.
Strong clinical and experimental evidence demonstrates association of elevated levels of matrix metalloproteinase MMP-9 with cancer progression, metastasis and shortened patient survival, as it plays a key role in tumor cell invasion and metastasis by digesting the basement membrane and ECM components. MMP-9 is secreted in both the monomeric and dimeric form. Although there is little research on MMP-9 dimers, some studies have shown the dimer to be associated with more aggressive tumor progression. Our objective was to study the relative secretion patterns of MMP-9 monomer and dimer in a variety of cancer cell lines and the effect of a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract on MMP-9 secretion. The cancer cell lines were grown in their respective media, supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 µg/ml) in 24-well tissue culture plates. At near confluence, the cells were treated with NM at 0,10, 50, 100, 500 and 1000 µg/ml. Parallel sets of cultures were treated with PMA (100 ng/ml) for induction of MMP-9. Cell MMP-9 secretion was assayed by gelatinase zymography. MMP-9 dimer secretion patterns of cancer cells fell into different categories. We observed no MMP-9 dimer in prostate DU-145 and PC-3, pancreatic MIA-Pa-Ca2, colon HCT-116, bladder T-24, head and neck FaDu, glioblastoma A-172, T-98 and LN-18 and leukemia HL-60, Jurkat, and Raji cell lines. MMP-dimer secretion only with PMA induction was seen in breast MCF-7 and MDA-MB-231, uterine SK-UT-1, lung A-549, tongue SC-25, melanoma A2058, osteosarcoma U-2OS, rhabdomyosarcoma, fibrosarcoma HT-1080, chondrosarcoma SW-1350 and liposarcoma SW-872. Cervical HeLa and DoTc 2 4510, renal 786-0 and HCC SK-Hep-1 cells exhibited MMP-9 dimer without PMA treatment and increased secretion with PMA treatment. Sarcomas had the highest levels of MMP-9 monomer and dimer with and without PMA among these cancer cell lines. Cervical, uterine and male breast cancer cell lines showed the next highest levels of MMP-9, followed by breast cancer cell lines. Melanoma, renal, lung, head and neck and HCC showed lower levels and prostate, glioblastoma, bladder and leukemia cell lines the lowest. NM showed dose-dependent inhibition of MMP-9 monomer and dimer in all cell lines tested. In conclusion, high MMP-9 and dimer secretion levels correlated with the most aggressive cancer cell lines. NM was effective in inhibiting MMP-9 and dimer secretion in all cell lines tested, suggesting its therapeutic potential as an antimetastatic agent.
大量临床和实验证据表明，基质金属蛋白酶 MMP-9 水平升高与癌症进展、转移和患者生存时间缩短有关，因为它在肿瘤细胞浸润和转移过程中通过消化基底膜和细胞外基质成分发挥关键作用。MMP-9 以单体和二聚体形式分泌。虽然对 MMP-9 二聚体的研究很少，但一些研究表明二聚体与更具侵袭性的肿瘤进展有关。我们的目的是研究各种癌细胞系中 MMP-9 单体和二聚体的相对分泌模式，以及含有赖氨酸、脯氨酸、抗坏血酸和绿茶提取物的营养混合物 (NM) 对 MMP-9 分泌的影响。将癌细胞系在各自的培养基中生长，在 24 孔组织培养板中用含有 10% FBS、青霉素 (100 U/ml) 和链霉素 (100 µg/ml) 的培养基补充。当接近汇合时，用 NM 在 0、10、50、100、500 和 1000 µg/ml 处理细胞。用 PMA(100 ng/ml) 处理平行培养物以诱导 MMP-9。通过明胶酶谱法测定细胞 MMP-9 分泌。癌细胞的 MMP-9 二聚体分泌模式分为不同类别。我们在前列腺 DU-145 和 PC-3、胰腺 MIA-Pa-Ca2、结肠 HCT-116、膀胱 T-24、头颈部 FaDu、胶质母细胞瘤 A-172、T-98 和 LN-18 和白血病 HL-60、Jurkat 和 Raji 细胞系中均未观察到 MMP-9 二聚体。仅在乳腺癌 MCF-7 和 MDA-MB-231、子宫 SK-UT-1、肺 A-549、舌 SC-25、黑色素瘤 A2058、骨肉瘤 U-2OS、横纹肌肉瘤、纤维肉瘤 HT-1080、软骨肉瘤 SW-1350 和脂肪肉瘤 SW-872 中观察到 MMP-二聚体仅在 PMA 诱导下分泌。宫颈 HeLa 和 DoTc 2 4510、肾 786-0 和 HCC SK-Hep-1 细胞在没有 PMA 处理的情况下表现出 MMP-9 二聚体，并在 PMA 处理后分泌增加。肉瘤在这些癌细胞系中具有最高水平的 MMP-9 单体和二聚体，无论是否有 PMA 存在。宫颈、子宫和男性乳腺癌细胞系显示出仅次于肉瘤的 MMP-9 水平最高，其次是乳腺癌细胞系。黑色素瘤、肾、肺、头颈部和 HCC 显示出较低的水平，前列腺、胶质母细胞瘤、膀胱和白血病细胞系显示出最低的水平。NM 对所有测试的细胞系中的 MMP-9 单体和二聚体均表现出剂量依赖性抑制作用。总之，高 MMP-9 和二聚体分泌水平与最具侵袭性的癌细胞系相关。NM 可有效抑制所有测试细胞系中的 MMP-9 和二聚体分泌，表明其作为抗转移剂的治疗潜力。
