Gene knockdown studies revealed CCDC50 as a candidate gene in mantle cell lymphoma and chronic lymphocytic leukemia
Authors:
Farfsing A, Engel F, Seiffert M, Hartmann E, Ott G, Rosenwald A, Stilgenbauer S, Döhner H, Boutros M, Lichter P, Pscherer A.
In:
Source:
Leukemia
Publication Date:
(
2009
)
Issue:
23(11)
:
2018-26
Research Area:
Cancer Research/Cell Biology
Immunotherapy / Hematology
Cells used in publication:
LCL
Species: human
Tissue Origin: blood
B-CLL
Species: human
Tissue Origin: blood
Granta519
Species: human
Tissue Origin: blood
MEC-2
Species: human
Tissue Origin: blood
JVM-2
Species: human
Tissue Origin: blood
Platform:
Nucleofector® I/II/2b
4D-Nucleofector® 96-well Systems
Experiment
Nufe 2b: primary B cells from patients with CLL: 5x10e6 cells/100µl, primary B cell solution; program U-015, 500nM siRNA MEC-1 and LCL-WEI cell lines: 2x10e6 cells/100µl; solution T, O-017; 500nM siRNA 96 well shuttle: JVM-2 and Granta cell line: 4 x 10e5 cells/20µl; Solution SF, program DN-100, 500nM siRNA
Abstract
The two B-cell non-Hodgkin's lymphoma entities, chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL), show recurrent chromosomal gains of 3q25-q29, 12q13-q14 and 18q21-q22. The pathomechanisms affected by these aberrations are not understood. The aim of this study was to identify genes, located within these gained regions, which control cell death and cell survival of MCL and CLL cancer cells. Blood samples collected from 18 patients with CLL and 6 patients with MCL, as well as 6 cell lines representing both malignancies were analyzed by gene expression profiling. By a comparison of genomic DNA and gene expression, 72 candidate genes were identified. We performed a limited RNA interference screening with these candidates to identify genes affecting cell survival. CCDC50 (coiled coil domain containing protein 50), SERPINI2 and SMARCC2 mediated a reduction of cell viability in primary CLL cells as well as in cell lines. Gene knockdown and a nuclear factor kappa B (NFkappaB) reporter gene assay revealed that CCDC50 is required for survival in MCL and CLL cells and controls NFkappaB signaling.
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