Effect of <i>Bh</i>L and <i>Di</i>L9 on activation of caspases.

<p>The lectin (<i>Bh</i>L: 0.247 and <i>Di</i>L9: 14.8 μM) treated PANC-1, CFPAC-1 and MIA PaCa-2 cells were incubated for different time periods and activities of caspases were assessed by fluorimetric assay. (A) Caspase-8, (B) caspase-9 and (C) caspase-3 activity was measured with respect to untreated cells. This data is mean ± SD values from three independent experiments.</p

Effect of <i>Bh</i>L and <i>Di</i>L9 on different phases of cell cycle.

<p>The progressive cell cycle changes were observed with DAPI staining after 6, 12, 18 and 24 h on lectin (<i>Bh</i>L: 0.247 and <i>Di</i>L9: 14.8 μM) treatment and analyzed using HCS software. The darkened numbers indicate the percentage of cells arrested in different phases of cell cycle. The data is mean ± SD of three independent experiments.</p

Inhibition of angiogenesis by <i>Bh</i>L and <i>Di</i>L9.

<p>HUVECs were plated on Matrigel (± LVES+VEGF) precoated 96-well plates as control. The cells were treated with lectins. The tubes were stained with rhodamine conjugated phalloidin (red) and nuclei with DAPI (blue). Anti-angiogenic activity is determined by the breakage in the tubule formation. Cell imaging was done on Cellomics’ ArrayScan HCS Reader.</p

<i>Bh</i>L and <i>Di</i>L9-induced apoptosis in PANC-1 cells.

<p>(A) Acridine orange staining of PANC-1 cells after incubation with lectins (<i>Bh</i>L: 0.247 and <i>Di</i>L9: 14.8 μM) for 24 h. The arrows indicate apoptotic bodies formed inside the lectin treated cells. Annexin V-FITC/PI staining of lectin stimulated PANC-1 cells incubated for indicated time. (B) The graph represents percentage of cells undergoing apoptosis, mean value ±SD of three independent experiments. (C) The overlay represents the cells that have undergone apoptosis (Annexin V-FITC positive, green) or necrosis (PI positive cells, red) after 72 h of incubation. The analysis was carried out using HCS 2.0 Cell Analysis Software.</p

Proposed model for mechanism of apoptosis induced by <i>Bh</i>L and <i>Di</i>L9 in PANC-1 cells.

<p>Lectins bind to the glycoprotein receptors of PANC-1 cells and triggers mitochondrial membrane depolarization cascade. The apoptotic signal was amplified by activation of caspase-9 and -3 leading to final cell death.</p

Effect of <i>Bh</i>L and <i>Di</i>L9 on proliferation and cell viability.

<p>Primary cells and different cancer cell lines were treated with serial concentrations of <i>Bh</i>L and <i>Di</i>L9 and incubated for 48 h. The growth inhibition (%) was measured by MTT assay by considering untreated cells as 100%. (A) Effect of <i>Bh</i>L and (B) <i>Di</i>L9 treatment on various cell lines. The values presented in the graph are the mean ± SD of two independent experiments done in triplicates.</p

Anti-proliferative effect of novel primary cetyl alcohol derived sophorolipids against human cervical cancer cells HeLa

<div><p>Sophorolipids (SLs) are glycolipid biosurfactants that have been shown to display anticancer activity. In the present study, we report anti-proliferative studies on purified forms of novel SLs synthesized using cetyl alcohol as the substrate (referred as SLCA) and their anticancer mechanism in human cervical cancer cells. Antiproliferative effect of column purified SLCA fractions (A, B, C, D, E and F) was examined in panel of human cancer cell lines as well as primary cells. Among these fractions, SLCA B and C significantly inhibited the survival of HeLa and HCT 116 cells without affecting the viability of normal human umbilical vein endothelial cells (HUVEC). The two fractions were identified as cetyl alcohol sophorolipids with non-hydroxylated tail differing in the degree of acetylation on sophorose head group. At an IC₅₀ concentration SLCA B (16.32 μg ml^-1) and SLCA C (14.14 μg ml^-1) blocked the cell cycle progression of HeLa cells at G1/S phase in time-dependent manner. Moreover, SLCA B and SLCA C induced apoptosis in HeLa cells through an increase in intracellular Ca²⁺ leading to depolarization of mitochondrial membrane potential and increase in the caspase-3, -8 and -9 activity. All these findings suggest that these SLCAs could be explored for their chemopreventive potential in cervical cancer.</p></div