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Anti-invasive gene expression profile of curcumin in lung adenocarcinoma based on a high throughput microarray
Huei-Wen Chen1, Sung-Liang Yu, Jeremy J W Chen
1Department of Medical Research, National Taiwan University Hospital, Taipei, Taiwan, ROC.
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 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.
Insights
Curcumin effectively reduces lung cancer cell invasion by downregulating key genes like MMP14, without significant toxicity. This natural compound shows promise as an antimetastatic agent by regulating gene expression to inhibit cancer spread.
Area of Science:
- Molecular Biology
- Cancer Research
- Pharmacology
Background:
- Curcumin is known for potential anti-invasive and antimetastatic properties.
- The precise mechanisms underlying curcumin's effects on cancer invasion are not fully understood.
- Lung adenocarcinoma cells (CL1-5) exhibit high invasiveness, making them a relevant model for study.
Purpose of the Study:
- To elucidate the anti-invasive mechanisms of curcumin in highly invasive lung adenocarcinoma cells.
- To identify specific genes and pathways regulated by curcumin that influence cellular invasion.
- To evaluate the concentration-dependent effects of curcumin on cell invasion and gene expression.
Main Methods:
- Microarray analysis of gene expression profiles to identify differentially expressed genes.
- Transwell cell culture chamber assay to quantify the invasive capacity of CL1-5 cells.
- Real-time quantitative reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry for validation.
- Gelatin zymographic analysis to assess matrix metalloproteinase (MMP) activity.
Main Results:
- Curcumin significantly reduced CL1-5 cell invasion in a concentration-dependent manner, below cytotoxic levels.
- Microarray analysis revealed downregulation of invasion-related genes, including matrix metalloproteinase 14 (MMP14) and integrins alpha6/beta4.
- Curcumin induced the expression of heat-shock proteins (Hsp27, Hsp70, Hsp40-like protein).
- Validated suppression of MMP14 at both mRNA and protein levels, and inhibition of MMP2 activity.
Conclusions:
- Curcumin exhibits significant anti-invasive effects on lung adenocarcinoma cells.
- The antimetastatic mechanism involves the downregulation of key invasion-related genes, notably MMP14.
- Curcumin may serve as a potential therapeutic agent for preventing cancer metastasis through gene expression modulation.