Anti-Invasive Gene Expression Profile of Curcumin in Lung Adenocarcinoma Based on a High Throughput Microarray Analysis

Abstract

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 M) and that this anti-invasive effect was concentration dependent (10.17 Ϯ 0.76 ϫ 10 3 cells at 0 M; 5.67 Ϯ 1.53 ϫ 10 3 cells at 1 M; 2.67 Ϯ 0.58 ϫ 10 3 cells at 5 M; 1.15 Ϯ 1.03 ϫ 10 3 cells at 10 M; P Ͻ 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 M), several invasion-related genes were suppressed, including matrix metalloproteinase 14 (MMP14; 0.65-fold), neuronal cell adhesion molecule (0.54-fold), and integrins ␣6 (0.67-fold) and ␤4 (0.63-fold). In addition, several heatshock proteins (Hsp) [Hsp27 (2.78-fold), Hsp70 (3.75-fold), and Hsp40-like protein (3.21-fold)] were induced by curcumin. Realtime quantitative reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry confirmed these results in both RNA and protein levels. Curcumin (1 to 10 M) 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.Curcumin, or diferuloylmethane, is a major chemical component of turmeric (Curcuma longa) and is used as a spice to give a specific flavor and yellow color to curry. It is also used as a cosmetic and in some medical preparations (Govindarajan, 1980). Curcumin has been shown to display anticarcinogenic properties in animals, as indicated by its ability to inhibit phorbol ester-induced skin tumors in a mouse model system (Huang et al., 1988). In addition to its anticarcinogenic effect, curcumin is also shown to exhibit anti-inflammatory, antiproliferative, antiangiogenic, and antioxidant properties . These effects of curcumin may be mediated by its inhibitory effect on a host of cell-signaling factors, including AP-1 transcription factor, c-Myc, Egr-1, NF-B, protein kinase C, epidermal growth factor receptor tyrosine kinase, c-Jun N-terminal kinase, protein tyrosine kinases, protein serine/threonine kinases, and IB kinase (Huang et al., 1988;Hong et al., 1999;Lin et al., 2000). In our previous study, it was also found that curcumin could inhibit cell cycle progression and induce cell apoptosis by regulating the gene expression of c-myc and bcl-2 (Chen and Huang, 1998).

Curcumin Restores Corticosteroid Function in Monocytes Exposed to Oxidants by Maintaining HDAC2

“…Moreover, the impact of 1 mM curcumin on gene expression was equally restricted. Unlike previously reported gene array data using higher concentrations of curcumin (60), no evidence of induction of Nrf2-responsive genes such as the antioxidant genes, or suppression of pro-inflammatory gene sets, was evident. This would help explain the limited impact that low concentrations of curcumin (, 1 mM) alone would have on ROSinduced inflammation, whether or not LPS was also present.…”

Curcumin Inhibits Lung Cancer Cell Invasion and Metastasis through the Tumor Suppressor HLJ1

“…The human lung carcinoma cell line, A549 [American Type Culture Collection (ATCC); CCL-185], was obtained from ATCC. The cells were grown in DMEM (Life Technologies) supplemented with 10% fetal bovine serum (FBS; Life Technologies) and each of penicillin and streptomycin (100 mg/mL each) at 37jC in a humidified atmosphere of 5% CO 2 (7).…”

“…This might be possible by synergistically modulating several of the properties of cancer such as proliferation, invasion, and metastasis. To provide a more global perspective, we examined the anticancer mechanisms of curcumin using cDNA microarray mRNA analysis of lung cancer cells and identified a panel of candidate curcumin target genes (7). Some of these target genes are highly associated with the cancer invasion and metastasis (8).…”

“…Several lines of evidence have shown that curcumin is able to inhibit cancer cell invasion and metastasis (3,7,20,21), and our previous microarray data suggested that not only the metalloproteinase (MMP) could be affected by curcumin, but also the tumor suppressor and invasion suppressor HLJ1 may be regulated by curcumin (7). In this study, we identified that the anti-invasive and antimetastasis effect of curcumin is mediated by up-regulation of HLJ1 and curcumin is able to transcriptionally regulated HLJ1 expression through the JNK/JunD pathway and inhibit lung cancer cell invasion and metastasis by modulating E-cadherin expression.…”

Ganoderic Acid Me Inhibits Tumor Invasion Through Down-Regulating Matrix Metalloproteinases 2/9 Gene Expression

“…The results showed that the cell viability of 95-D cells at 0, 5, 10, and 20 μg/ ml of GA-Me was 98.3 ± 2.9%, 96.4 ± 0.6%, 93.1 ± 2.3%, and 92.4 ± 1.8%, respectively (data not shown). Apoptosis of tumor cells was only 2.3% at 20 μg / ml of GA-Me for 24 h. These results can be compared to the reported data on curcumin: Exposure of CL1-5 cells, which have high invasive capacities, to curcumin (0, 1, 5, 10, 20 μM) influenced the cell viability of CL1-5 cells in a concentration-dependent manner (92.3 ± 6.1% at 10 μM and 70.1 ± 7.3% at 20 μM), and curcumin (1 -20 μM) could significantly inhibit cell invasion of CL1-5 cells (56.0 ± 16.1% at 1.0 μM) (13).…”