Schematic preparation of carbazole ECAP in Cl₃/Ethanol, reflux, 5 h RT.

<p>Schematic preparation of carbazole ECAP in Cl₃/Ethanol, reflux, 5 h RT.</p

Characterization of the novel (Z)-4-((9-ethyl-9H-carbazol-3-yl) amino) pent-3-en-2-one (ECAP).

<p>(A) IR, (B) ¹H-NMR and (C) ¹³C-NMR spectrums.</p

Time dependent effect of ECAP on caspase activity and ATP level.

<p>(A) The activity of Caspase-3/7 (6 h: p < 0.4036, 24 h: p < 0.0003), (B) Caspase-8 (6 h: p < 0.4364, 24 h: p < 0.0001), (C) Caspase-9 (6 h: 0.4171, 24 h: p < 0.0124) and (D) ATP (6 h: 0.4011, 24 h: 0.0011) levels in A549 lung cancer cell line after 6 h and 24 h incubation. [* denotes statistical significance with respect to the control and uncertainties represent standard deviation (SD) from the means. Number of replicates: n ≥ 3].</p

The effect of ECAP on the induction of apoptosis of A549 cells after 24 h treatment.

<p>[(p < 0.0001), *** significance compared to the control and number of replicates: n ≥ 3].</p

Comet assay showing DNA damage in A549 cells after 24 h treatment.

<p>(A) Control and (B) ECAP treated A549 cells. (p < 0.0001).</p

Cytotoxic Effect of a Novel Synthesized Carbazole Compound on A549 Lung Cancer Cell Line

<div><p>Increased death rates due to lung cancer have necessitated the search for potential novel anticancer compounds such as carbazole derivatives. Carbazoles are aromatic heterocyclic compounds with anticancer, antibacterial and anti-inflammatory activity. The study investigated the ability of the novel carbazole compound (Z)-4-[9-ethyl-9aH-carbazol-3-yl) amino] pent-3-en-2-one (ECAP) to induce cytotoxicity of lung cancer cells and its mechanism of action. ECAP was synthesized as a yellow powder with melting point of 240-247 °C. The 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lipid peroxidation and comet assays were used to assess the cytotoxic effect of the compound on A549 lung cancer cells. Protein expression was determined using western blots, apoptosis was measured by luminometry (caspase-3/7, -8 and -9) assay and flow cytometry was used to measure phosphatidylserine (PS) externalisation. ECAP induced a p53 mediated apoptosis of lung cancer cells due to a significant reduction in the expression of antioxidant defence proteins (Nrf2 and SOD), Hsp70 (p < 0.02) and Bcl-2 (p < 0.0006), thereby up-regulating reactive oxygen species (ROS) production. This resulted in DNA damage (p < 0.0001), up-regulation of Bax expression and caspase activity and induction of apoptosis in lung cancer cells. The results show the anticancer potential of ECAP on lung cancer.</p></div

The effect of ECAP on lipid peroxidation on A549 lung cancer cells after 24 h incubation.

<p>(p < 0.0002) [*** significance compared to the control, number of replicates: n ≥ 3].</p

A schematic summary of apoptotic pathway of a novel carbazole compound (Z)-4-[9-ethyl-9aH-carbazol-3-yl) amino] pent-3-en-2-one on A549 lung cancer cells.

<p>A schematic summary of apoptotic pathway of a novel carbazole compound (Z)-4-[9-ethyl-9aH-carbazol-3-yl) amino] pent-3-en-2-one on A549 lung cancer cells.</p

IC₅₀ values of ECAP and ellipticine on A549 lung cancer cell line after 24 h treatment.

<p>N = 3 replicates at 95% confidence interval</p><p>IC₅₀ values of ECAP and ellipticine on A549 lung cancer cell line after 24 h treatment.</p

Western blot data shwing effect of the novel ECAP on protein expression in A549 lung cancer cell line.

<p>(A) Bax, (B) Bcl-2, (C) p53, (D) Nrf2, (E) Hsp70 and (F) SOD.</p