Cytotoxic effect of a novel synthesized carbazole compound on A549 lung cancer cell line

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), upregulation 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.S1 Fig. MTT assay measured in untreated (control) and 1% DMSO (vehicle control) treated A549 cells. The data showed no significant cytotoxicity.S2 Fig.Western blots showing the effect of ECAP on the expression of Bax, Bcl-2, p53, Nrf2, Hsp70 and SOD. The original western blots which were used for western blot analysis (Fig 6).College of Health Sciences, University of KwaZulu-Natalhttp://www.plosone.orgam201

Convenient and Efficient Microwave-Assisted Synthesis of a Methyl Derivative of the Fused Indoloquinoline Alkaloid Cryptosanguinolentine

An efficient synthesis of a methyl derivative of the indoloquinoline alkaloid cryptosanguinolentine based on microwave-assisted reactions is described. The microwave-assisted synthesis of an intermediate 4-hydroxy-2-methylquinoline yielded 86% of the desired product and other intermediates prepared yielded high % of products in shorter reaction times, under optimum conditions, as compared to traditional methods

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

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

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

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

Enhanced catalytic conversion of palm oil into biofuels by Cr-incorporated sulphated zirconia

The conversion of palm oil into biofuels using commercial zirconia (ZrO2) modified sulfuric acid (H2SO4) incorporated with Cr metal has been systematically studied. Sulphated zirconia (ZrO2-SO4) was prepared by dispersing ZrO2 in H2SO4 solution via wet impregnation method. Cr-incorporated sulphated zirconia (Cr/ZrO2-SO4) were prepared hydrothermally through wet impregnation of ZrO2-SO4 and chromium (III) nitrate nonahydrate (Cr(NO3)3·9H2O) solution. The results showed that reduction step with pure hydrogen gas (H2) over non-calcined catalyst (Cr/ZrO2-SO4 (NC)) resulted in higher activity and selectivity than calcined catalyst (Cr/ZrO2-SO4 (C600)). A maximum acidity test result was obtained on Cr/ZrO2-SO4 (NC) of 2.33 mmol g−1, increasing from Cr/ZrO2-SO4 (C600) of 1.32 mmol g−1. The selectivity of gasoline fraction demonstrated by Cr/ZrO2-SO4 (C600) and Cr/ZrO2-SO4 (NC) was observed to contain 35.85% and 36.51%, respectively