Roomi M W, Monterrey J C, Kalinovsky T, Rath M, Niedzwiecki A
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050, USA.
Exp Oncol. 2010 Dec;32(4):243-8.
Matrix metalloproteinase (MMP)-2 and -9 secretion is elevated in various human cancers and their elevated expression has been associated with poor prognosis due to associated increased cancer cell invasion and metastasis.
To examine the correlation between in vitro MMP-2 and MMP-9 secretion and Matrigel invasion in 42 different human cancer cell lines (selected on the basis of organ malignancies) treated with a nutrient mixture (NM).
The cells were cultured in their recommended media supplemented with 10% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were treated with NM dissolved in media at 0, 10, 50, 100, 500 and 1000 μg/mL in triplicate. Parallel sets of cultures were also treated with phorbol 12-myristate 13-acetate (PMA) 100 ng/mL for induction of enzymes. After 24 h the media were collected and MMP-2 and MMP-9 levels were assayed by gelatinase zymography. Invasion studies were conducted using Matrigel in 24-well plates.
Correlation of pooled data from different cancer cell line groups demonstrated dose-dependent inhibition of MMP-2 and -9 and Matrigel invasion with NM treatment and significant negative correlation between MMP-2 and MMP-9 levels and Matrigel invasion. Pooled data of cell lines expressing only MMP-2 and resistance to PMA induction of MMP-9 showed significant negative correlation (r = -0.77, p = 0.003) between MMP-2 secretion and inhibition of invasion through Matrigel. Cell lines expressing only MMP-9, showed significant negative correlation (r = -0.726, p = 0.003) between MMP-9 secretion and Matrigel invasion. Pooled data of cell lines expressing MMP-2 and MMP-9 demonstrated significant negative correlation (r = -0.821, p < 0.0001) between MMP-9 secretion and inhibition of invasion through Matrigel. Pooled data of cancer cell lines expressing no basal MMP- 9 secretion demonstrated significant negative correlation (r = -0.686, p < 0.0001) between PMA-induced MMP-9 secretion and inhibition of invasion through Matrigel.
In conclusion, regardless of MMP-2 and MMP-9 patterns of expression, MMP modulation by NM was found to be significantly correlated with NM modulation of Matrigel invasion of these cell lines.
基质金属蛋白酶（MMP）-2和-9在多种人类癌症中的分泌增加，其表达升高与不良预后相关，因为癌细胞侵袭和转移增加。
研究在42种不同的人类癌细胞系（根据器官恶性肿瘤选择）中，用营养混合物（NM）处理后，体外MMP-2和MMP-9分泌与基质胶侵袭之间的相关性。
将细胞接种于24孔组织培养板中，在其推荐培养基中添加10%胎牛血清和抗生素进行培养。接近汇合时，将细胞分别用溶解于培养基中的NM处理，浓度为0、10、50、100、500和1000μg/mL，一式三份。平行培养组还用100 ng/mL佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）处理以诱导酶的产生。24小时后收集培养基，通过明胶酶谱法检测MMP-2和MMP-9水平。使用24孔板中的基质胶进行侵袭研究。
来自不同癌细胞系组的汇总数据显示，NM处理对MMP-2和-9以及基质胶侵袭具有剂量依赖性抑制作用，且MMP-2和MMP-9水平与基质胶侵袭之间存在显著负相关。仅表达MMP-2且对PMA诱导MMP-9有抗性的细胞系汇总数据显示，MMP-2分泌与通过基质胶的侵袭抑制之间存在显著负相关（r = -0.77，p = 0.003）。仅表达MMP-9的细胞系显示，MMP-9分泌与基质胶侵袭之间存在显著负相关（r = -0.726，p = 0.003）。表达MMP-2和MMP-9的细胞系汇总数据显示，MMP-9分泌与通过基质胶的侵袭抑制之间存在显著负相关（r = -0.821，p < 0.0001）。不分泌基础MMP-9的癌细胞系汇总数据显示，PMA诱导的MMP-9分泌与通过基质胶的侵袭抑制之间存在显著负相关（r = -0.686，p < 0.0001）。
总之，无论MMP-2和MMP-9的表达模式如何，发现NM对MMP的调节与这些细胞系基质胶侵袭的NM调节显著相关。
