Yodkeeree Supachai, Garbisa Spiridione, Limtrakul Pornngarm
Department of Biochemistry, Chiang Mai University, Chiang Mai, Thailand.
Acta Pharmacol Sin. 2008 Jul;29(7):853-60. doi: 10.1111/j.1745-7254.2008.00792.x.
Tetrahydrocurcumin (THC) is an active metabolite of curcumin. It has been reported to have similar pharmacological activity to curcumin. The proteases that participate in extracellular matrix (ECM) degradation are involved in cancer cell metastasis. The present study investigates the effect of an ultimate metabolite of curcumin, THC, on the invasion and motility of highly-metastatic HT1080 human fibrosarcoma cells.
The effect of THC on HT1080 cell invasion and migration was determined using Boyden chamber assay. Cell-adhesion assay was used for examining the binding of cells to ECM molecules. Zymography assay was used to analyze the effect of THC on matrix metalloproteinase (MMP)-2, MMP-9, and urokinase plasminogen activator (uPA) secretion from HT1080 cells. Tissue inhibitor of metalloproteinase (TIMP)-2 and membrane-type 1 matrix metalloproteinase (MT1-MMP) proteins levels were analyzed by Western blotting.
Treatment with THC reduced HT1080 cell invasion and migration in a dose-dependent manner. THC also decreased the cell adhesion to Matrigel and laminin-coated plates. Analysis by zymography demonstrated that treatment with THC reduced the levels of MMP-2, MMP-9, and uPA. THC also inhibited the levels of MT1-MMP and TIMP-2 proteins detected by Western blot analysis.
Our findings revealed that THC reduced HT1080 cell invasion and migration. The inhibition of cancer cell invasion is associated with the downregulation of ECM degradation enzymes and the inhibition of cell adhesion to ECM proteins.
四氢姜黄素（THC）是姜黄素的一种活性代谢产物。据报道，它具有与姜黄素相似的药理活性。参与细胞外基质（ECM）降解的蛋白酶与癌细胞转移有关。本研究调查了姜黄素的最终代谢产物THC对高转移性人纤维肉瘤HT1080细胞侵袭和运动能力的影响。
使用Boyden小室分析法测定THC对HT1080细胞侵袭和迁移的影响。细胞黏附试验用于检测细胞与ECM分子的结合。酶谱分析法用于分析THC对HT1080细胞基质金属蛋白酶（MMP）-2、MMP-9和尿激酶型纤溶酶原激活剂（uPA）分泌的影响。通过蛋白质印迹法分析金属蛋白酶组织抑制剂（TIMP）-2和膜型1基质金属蛋白酶（MT1-MMP）的蛋白水平。
THC处理以剂量依赖的方式降低了HT1080细胞的侵袭和迁移能力。THC还降低了细胞与基质胶和层粘连蛋白包被平板的黏附。酶谱分析表明，THC处理降低了MMP-2、MMP-9和uPA的水平。THC还抑制了蛋白质印迹分析检测到的MT1-MMP和TIMP-2蛋白水平。
我们的研究结果表明，THC降低了HT1080细胞的侵袭和迁移能力。对癌细胞侵袭的抑制与ECM降解酶的下调以及细胞对ECM蛋白黏附的抑制有关。
