Chen Huei-Wen, Yu Sung-Liang, Chen Jeremy J W, Li Han-Ni, Lin Yi-Chen, Yao Pei-Li, Chou Han-Yi, Chien Chiang-Ting, Chen Wen-Jone, Lee Yuan-Teh, Yang Pan-Chyr
Department of Medical Research, National Taiwan University Hospital, Taipei, Taiwan, ROC.
Mol Pharmacol. 2004 Jan;65(1):99-110. doi: 10.1124/mol.65.1.99.
Curcumin has been reported to exhibit anti-invasive and/or antimetastatic activities, but the mechanism remains unclear. In this study, microarray analysis of gene expression profiles were used to characterize the anti-invasive mechanisms of curcumin in highly invasive lung adenocarcinoma cells (CL1-5). Results showed that curcumin significantly reduces the invasive capacity of CL1-5 cells in a concentration range far below its levels of cytotoxicity (20 microM) and that this anti-invasive effect was concentration dependent (10.17 +/- 0.76 x 10(3) cells at 0 microM; 5.67 +/- 1.53 x 10(3) cells at 1 microM; 2.67 +/- 0.58 x 10(3) cells at 5 microM; 1.15 +/- 1.03 x 10(3) cells at 10 microM; P < 0.05) in the Transwell cell culture chamber assay. Using microarray analysis, 81 genes were down-regulated and 71 genes were up-regulated after curcumin treatment. Below sublethal concentrations of curcumin (10 microM), several invasion-related genes were suppressed, including matrix metalloproteinase 14 (MMP14; 0.65-fold), neuronal cell adhesion molecule (0.54-fold), and integrins alpha6 (0.67-fold) and beta4 (0.63-fold). In addition, several heat-shock proteins (Hsp) [Hsp27 (2.78-fold), Hsp70 (3.75-fold), and Hsp40-like protein (3.21-fold)] were induced by curcumin. Real-time quantitative reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry confirmed these results in both RNA and protein levels. Curcumin (1 to 10 microM) reduced the MMP14 expression in both mRNA and protein levels and also inhibited the activity of MMP2, the down-stream gelatinase of MMP14, by gelatin zymographic analysis. Based on these data, it can be concluded that curcumin might be an effective antimetastatic agent with a mechanism of anti-invasion via the regulation of certain gene expressions.
姜黄素已被报道具有抗侵袭和/或抗转移活性，但其机制仍不清楚。在本研究中，利用基因表达谱的微阵列分析来表征姜黄素在高侵袭性肺腺癌细胞（CL1-5）中的抗侵袭机制。结果显示，姜黄素在远低于其细胞毒性水平（20微摩尔）的浓度范围内显著降低CL1-5细胞的侵袭能力，且这种抗侵袭作用呈浓度依赖性（在Transwell细胞培养小室分析中，0微摩尔时为10.17±0.76×10³个细胞；1微摩尔时为5.67±1.53×10³个细胞；5微摩尔时为2.67±0.58×10³个细胞；10微摩尔时为1.15±1.03×10³个细胞；P<0.05）。通过微阵列分析，姜黄素处理后有81个基因下调，71个基因上调。在低于姜黄素亚致死浓度（10微摩尔）时，包括基质金属蛋白酶14（MMP14；0.65倍）、神经细胞黏附分子（0.54倍）以及整合素α6（0.67倍）和β4（0.63倍）在内的几个侵袭相关基因受到抑制。此外，姜黄素诱导了几种热休克蛋白（Hsp）[Hsp27（2.78倍）、Hsp70（3.75倍）和类Hsp40蛋白（3.21倍）]。实时定量逆转录-聚合酶链反应、蛋白质印迹法和免疫组织化学在RNA和蛋白质水平上均证实了这些结果。姜黄素（1至10微摩尔）在mRNA和蛋白质水平上均降低了MMP14的表达，并且通过明胶酶谱分析抑制了MMP14的下游明胶酶MMP2的活性。基于这些数据，可以得出结论，姜黄素可能是一种有效的抗转移剂，其抗侵袭机制是通过调节某些基因表达实现的。
