Roomi M Waheed, Ivanov Vadim, Kalinovsky Tatiana, Niedzwiecki Aleksandra, Rath Matthias
Cancer Division, Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA, 95050, USA.
Med Oncol. 2007;24(2):183-8. doi: 10.1007/BF02698038.
Current treatment of testicular cancer is associated with secondary malignancy, infertility, and cytotoxicity. Based on reported antimetastatic potential, we investigated the effect of a nutrient mixture (NM) containing lysine, proline, arginine, ascorbic acid, and green tea extract on human testis cancer cell line NT 2/DT by measuring cell proliferation/cytotoxicity, modulation of MMP-2 and MMP-9 secretion, and cancer cell invasive potential. Human testis cancer cells NT 2/DT (ATCC) were grown in DME medium. At near confluence, the cells were treated with NM dissolved in media and tested at 0,10, 50, and 100 microg/mL in triplicate at each dose. Cells were also treated with PMA 200 ng/mL to study enhanced secretion of MMP-9. Cell proliferation/cytotoxicity was evaluated by MTT assay, MMP activity by gelatinase zymography, and invasion through Matrigel. The nutrient mixture showed no significant effect on testis cancer cell growth. Zymography demonstrated secretion of MMP-2 by untreated human testis cancer cells and MMP-9 with PMA induction. NM inhibited secretion of both MMPs in a dose-dependent fashion with virtual total inhibition of MMP-9 at 100 microg/mL. Invasion of human testis cancer cells through Matrigel was reduced by 84% at 50 microg/mL and at 100 microg/mL (p = 0.004). NM significantly inhibited MMP secretion and matrix invasion in testicular cancer cells without toxic effect, indicating potential as an anticancer agent.
目前睾丸癌的治疗会引发继发性恶性肿瘤、不育症以及细胞毒性。基于已报道的抗转移潜力,我们通过测量细胞增殖/细胞毒性、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)分泌的调节以及癌细胞侵袭潜力,研究了一种包含赖氨酸、脯氨酸、精氨酸、抗坏血酸和绿茶提取物的营养混合物(NM)对人睾丸癌细胞系NT 2/DT的影响。人睾丸癌细胞NT 2/DT(美国典型培养物保藏中心)在杜氏改良 Eagle 培养基(DME)中培养。接近汇合时,用溶解于培养基中的NM处理细胞,并在每个剂量下以0、10、50和100微克/毫升进行一式三份测试。还用200纳克/毫升的佛波酯(PMA)处理细胞,以研究MMP-9分泌的增强情况。通过MTT法评估细胞增殖/细胞毒性,通过明胶酶谱法评估MMP活性,通过基质胶评估侵袭情况。该营养混合物对睾丸癌细胞生长无显著影响。酶谱分析表明,未处理的人睾丸癌细胞分泌MMP-2,经PMA诱导后分泌MMP-9。NM以剂量依赖方式抑制这两种MMP的分泌,在100微克/毫升时对MMP-9几乎完全抑制。在50微克/毫升和100微克/毫升时,人睾丸癌细胞通过基质胶的侵袭减少了84%(p = 0.004)。NM显著抑制睾丸癌细胞中的MMP分泌和基质侵袭,且无毒性作用,表明其具有作为抗癌剂的潜力。