Roomi M W, Kalinovsky T, Roomi N M, Cha J, Rath M, Niedzwiecki A
Dr. Rath Research Institute, Santa Clara, CA 95050, USA.
Int J Oncol. 2014 Jun;44(6):1933-44. doi: 10.3892/ijo.2014.2379. Epub 2014 Apr 10.
Long-term survival of patients with breast cancer remains poor, due to metastasis and recurrence. We investigated the effects of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract in vitro and in vivo on 4T1 murine breast cancer, a representative model for metastatic breast cancer. After one week of isolation, 5-6-week-old female Balb/C mice were inoculated with 5x10⁵ 4T1 cells into the mammary pad and randomly divided into two groups; the control group was fed a regular diet and the NM group a regular diet supplemented with 0.5% NM. After four weeks, the mice were sacrificed and their tumors, lungs, livers, kidneys, hearts and spleens were excised and processed for histology. Dimensions (length and width) of tumors were measured using a digital caliper, and the tumor burden was calculated using the following formula: 0.5 x length x width. We also tested the effect of NM in vitro on 4T1 cells, measuring cell proliferation by MTT assay, MMP secretion by zymography, invasion through Matrigel, migration by scratch test and morphology by H&E staining. NM inhibited tumor weight and burden of 4T1 tumors by 50% (p=0.02) and 53.4% (p≤0.0001), respectively. Lung metastasis was profoundly inhibited by NM supplementation: mean number of colonies was reduced by 87% (p<0.0001) and mean weight of lungs by 60% (p=0.0001) compared to control mice. Metastasis to liver, spleen, kidney and heart was significantly reduced with NM supplementation. In vitro, NM exhibited 50% toxicity over the control at 250 and 500 µg/ml concentrations. Zymography demonstrated MMP-2 and MMP-9 secretion which was inhibited by NM in a dose-dependent manner, with virtual total inhibition of both at 1,000 µg/ml. Migration by scratch test and invasion through Matrigel were inhibited in a dose-dependent manner with total block of invasion at 250 and of migration at 1,000 µg/ml. These results suggest that NM has therapeutic potential in the treatment of breast cancer.
由于转移和复发，乳腺癌患者的长期生存率仍然很低。我们研究了一种含有抗坏血酸、赖氨酸、脯氨酸和绿茶提取物的新型营养混合物（NM）在体外和体内对4T1小鼠乳腺癌（转移性乳腺癌的代表性模型）的影响。隔离一周后，将5 - 6周龄的雌性Balb/C小鼠在乳腺垫中接种5×10⁵个4T1细胞，并随机分为两组；对照组给予常规饮食，NM组给予补充了0.5% NM的常规饮食。四周后，处死小鼠，切除其肿瘤、肺、肝、肾、心脏和脾脏并进行组织学处理。使用数字卡尺测量肿瘤的尺寸（长度和宽度），并使用以下公式计算肿瘤负荷：0.5×长度×宽度。我们还在体外测试了NM对4T1细胞的作用，通过MTT法测量细胞增殖，通过酶谱法测量MMP分泌，通过基质胶测量侵袭，通过划痕试验测量迁移，并通过苏木精和伊红染色观察形态。NM分别使4T1肿瘤的重量和负荷降低了50%（p = 0.02）和53.4%（p≤0.0001）。补充NM可显著抑制肺转移：与对照小鼠相比，集落平均数减少了87%（p < 0.0001），肺平均重量减少了60%（p = 0.0001）。补充NM后，肝、脾、肾和心脏的转移也显著减少。在体外，NM在250和500μg/ml浓度下对对照组表现出50%的毒性。酶谱法显示MMP - 2和MMP - 9的分泌受到NM的剂量依赖性抑制，在1000μg/ml时两者几乎完全被抑制。划痕试验的迁移和通过基质胶的侵袭受到剂量依赖性抑制，在250μg/ml时侵袭完全被阻断，在1000μg/ml时迁移完全被阻断。这些结果表明NM在乳腺癌治疗中具有治疗潜力。
