Figure 1.
(A, B, C) Effect of the diatom PUA 2-trans,4-trans-decadienal (DD) on the human lung adenocarcinoma cell lines A549 and COLO 205, and the lung/brunch normal epithelial BEAS-2B cell line.
(D) Effect of DD on A549 cell line in the presence of caspase-3 inhibitor (9.7 µM). Percentage of viable cells for A549 and COLO 205 calculated with the Trypan blue viability assay and for BEAS-2B with the MTT viability assay. Values are reported as mean ±S.D compared to controls (100% viability); ▴ 2 µM; ▪ 5 µM, ♦ 10 µM.
Figure 2.
(A, B, C) Effect of the diatom PUA 2-trans,4-trans-octadienal (OD) on the human lung adenocarcinoma cell lines A549 and COLO 205, and the lung/brunch normal epithelial BEAS-2B cell line.
(D) Effect of OD on A549 cell line in the presence of caspase-3 inhibitor (9.7 µM). Percentage of viable cells for A549 and COLO 205 calculated with the Trypan blue viability assay and for BEAS-2B with the MTT viability assay. Values are reported as mean ±S.D compared to controls (100% viability); ▴ 2 µM; ▪ 5 µM, ♦ 10 µM.
Figure 3.
(A, B, C) Effect of the diatom PUA 2-trans,4-trans-heptadienal (HD) on the human lung adenocarcinoma cell lines A549 and COLO 205, and the lung/brunch normal epithelial BEAS-2B cell line.
(D) Effect of HD on A549 cell line in the presence of caspase-3 inhibitor (9.7 µM). Percentage of viable cells for A549 and COLO 205 calculated with the Trypan blue viability assay and for BEAS-2B with the MTT viability assay. Values are reported as mean ±S.D compared to controls (100% viability); ▴ 2 µM; ▪ 5 µM, ♦ 10 µM.
Figure 4.
Effect of the diatom PUAs 2-trans,4-trans-decadienal (DD), 2-trans,4-trans-octadienal (OD) and 2-trans,4-trans-heptadienal (HD) on the human lung adenocarcinoma cell line A549.
Control and treated cells double stained with acridine orange and ethidium bromide after 48 µM DD, OD and HD observed at the confocal microscope. Numbers indicate (1) normal cells; (2) early apoptotic cells; (3) late apoptotic cells; (4) necrotic cells (see material and methods for details). Arrows indicate cells with fragmented nuclei.
Figure 5.
The histograms show the effects of 2-trans,4-trans-decadienal (DD), 2-trans,4-trans-octadienal (OD) and 2-trans,4-trans-heptadienal (HD) on the expression levels of target proteins (TNFR2, TNFR1, Fas, FADD, RIP, caspase-3) and control (actin) in lung adenocarcinoma A549 cells.
Immunoblot analysis shows that PUAs induce TNF signaling after 24(DD and HD) and 48 h (OD) of treatment with actin. Asterisk denotes significant increase in protein levels measured. **p≤0.05 versus control; error bars represent ±SD.
Figure 6.
Histograms show the effects of 2-trans,4-trans-decadienal (DD) and 2-trans,4-trans-heptadienal (HD) on the expression levels of target genes in lung adenocarcinoma A549 cells.
Gene expression analysis was conducted after 2 µM DD and HD; error bars represent ±SD.
Figure 7.
(A) Flow cytometric analysis of DNA content.
Cells were exposed to 2-trans,4-trans-heptadienal (HD) 5 µM for 24 h. (B), The corresponding percentage of cells in each phase, obtained by ModFit LT Software.
Table1.
Percetage of A549 cells treated with the PUAs 2-trans,4-trans-decadienal (DD), 2-trans,4-trans-octadienal (OD) and 2-trans,4-trans-heptadienal (HD) in each cell cycle phase after times 0 and 24 h.
Figure 8.
Schematic representation of pathways induced by 2-trans,4-trans-decadienal (DD), 2-trans,4-trans–octadienal (OD) and 2-trans,4-trans-heptadienal (HD) in A549 cell line.
Black arrows indicate possible correlated pathways not involved in the response of cells to aldehyde treatment.