An article published this year in “Oncotarget” using one of our products, “PE
Apoptosis Detection Kit”,
by our customers from BioCruces Health Research Institute, Bizkaia,
Spain, in the analysis of how Podocalyxin promotes proliferation and survival
in mature B-cell non-Hodgkin lymphoma cells. Congrats and Thanks.
Summay:
Podocalyxin (PCLP1) is a CD34-related
sialomucin expressed by some normal cells and a variety of malignant tumors,
including leukemia, and associated with the most aggressive cancers and poor
clinical outcome. PCLP1 increases breast tumor growth, migration and invasion;
however, its role in hematologic malignancies still remains undetermined. The
purpose of this study was to investigate the expression and function of PCLP1
in mature B-cell lymphoma cells. We found that overexpression of PCLP1
significantly increases proliferation, cell-to-cell interaction, clonogenicity,
and migration of B-cell lymphoma cells. Furthermore, PCLP1 overexpression
results in higher resistance to death induced by dexamethasone, reactive oxygen
species and type II anti-CD20 monoclonal antibody obinutuzumab. Strikingly,
enforced expression of PCLP1 enhances lipid droplet formation as well as
pentose phosphate pathway and glutamine dependence, indicative of metabolic
reprogramming necessary to support the abnormal proliferation rate of tumor
cells. Flow cytometry analysis revealed augmented levels of PCLP1 in malignant
cells from some patients with mature B-cell lymphoma compared to their normal
B-cell counterparts. In summary, our results demonstrate that PCLP1 contributes
to proliferation and survival of mature B-cell lymphoma cells, suggesting that
PCLP1 may promote lymphomagenesis and represents a therapeutic target for the
treatment of B-cell lymphomas.
A) Raji-Ctrl and Raji-PCLP1 cells were treated with increasing concentrations of vehicle or dexamethasone for 72 hours and cell death was analyzed after Annexin V-PE/7´AAD staining by flow cytometry. The graph on the left shows the mean percentages ± SD of total specific cell death (including early apoptosis, late apoptosis and necrotic cells) from six independent experiments after subtracting the spontaneous cell death (vehicle). The percentage of specific cell death was calculated as follows: [(% lysis of target cell – % spontaneous cell death)/(100% – % spontaneous cell death)] × 100. Representative dot plots showing Annexin V/7AAD staining in cells treated with vehicle or 1000 µM dexamethasone are shown (right). As displayed in the dot plots, the spontaneous cell death (Annexin V PE positive/7ADD negative, Annexin V PE positive/7ADD positive and Annexin V PE negative/7ADD positive cells) in cells cultured with the vehicle was approximately 7.5% in Raji-Ctrl and 5.5% in Raji-PCLP1 cells. (B) Expression of PCLP1 on Annexin V and 7ADD double-negative population of cells treated with dexamethasone or vehicle by flow cytometry. (C) Raji-Ctrl and Raji-PCLP1 cells were treated with increasing concentrations of vehicle or H2O2 for 6 h and cell death was analyzed as indicated in A. The graph on the left shows the mean percentages ± SD of total cell death from five independent experiments after subtracting the spontaneous cell death (vehicle). Representative dot plots showing Annexin V/7AAD staining in cells treated with vehicle or 200 µM H2O2 are shown (left). As displayed in the dot plots, the spontaneous cell death in the presence of vehicle was approximately 17.0% in Raji-Ctrl and 5.9% in Raji-PCLP1 cells. (D) Cells were incubated overnight with increasing concentrations of obinutuzumab or vehicle and cell viability was measured based on Annexin V/7AAD double negative staining relative to vehicle-treated cells. Data show the mean percentages ± SD of viable cells from six independent experiments. (E) A representative image of the homotypic adhesion induced by obinutuzumab as visualized by light microscopy is shown. The size bar indicates 100 µm. Statistical analysis was calculated using two-tailed paired Student´s t-test. *P < 0.05. **P < 0.01. ***P < 0.001.
Reference:
Product link:
PE Apoptosis
Detection Kit
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