Dysregulation of the receptor activator of NF-kappaB ligand and osteoprotegerin production influence the apoptosis of multiple myeloma patients' bone marrow stromal cells co-cultured with myeloma cells.
The interaction of multiple myeloma (MM) cells and bone marrow stromal cells (BMSCs) induces profound changes in the bone marrow environment, influencing osteoclastogenesis and MM cell survival. Differences in receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG) production in BMSCs derived from MM patients and control subjects and the apoptosis of BMSCs and MM cells in co-cultures of both cell types were examined. RANKL and OPG expressions were examined by ELISA and semiquantitative RT-PCR. Apoptosis of BMSCs after contact with RPMI8226 and U266 cells was measured by flow cytometry and the level of ALP activity by the spectrophotometric method. OPG production by BMSCs was significantly inhibited after direct contact with RPMI8226 cells. Production of soluble RANKL was enhanced and the increase was more significant in the BMSCs of the MM patients than in those of the controls. In co-cultures of BMSCs and MM cells, significant apoptosis was detected with a concomitant decrease in ALP activity. This apoptosis decreased significantly in the presence of RANK-Fc, an antagonist of RANKL. Disturbances in the RANKL/OPG system are more profound in the BMSCs of MM patients than in those of control subjects after direct contact with RPMI8226 cells. Moreover, direct contact with RPMI8226 and U266 cells induces apoptosis of BMSCs which is mediated by an overproduction of RANKL.
Department of Virology and Immunology, Maria Curie-Skłodowska University, 20-033 Lublin, Poland. basiaz@poczta.umcs.lublin.pl
Barbara
B
Zdzisińska
Agnieszka
A
Bojarska-Junak
Adam
A
Walter-Croneck
Martyna
M
Kandefer-Szerszeń
D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000378:metabolism* / Q000473:pathology; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008875:Middle Aged; D009101:Multiple Myeloma / Q000378:metabolism* / Q000473:pathology; D053244:Osteoprotegerin / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D053245:RANK Ligand / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013053:Spectrophotometry; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology; D013997:Time Factors
D053244:Osteoprotegerin; D053245:RANK Ligand; C517736:Rank-Fc; D011993:Recombinant Fusion Proteins; C506030:TNFRSF11B protein, human; C505688:TNFSF11 protein, human; D000469:Alkaline Phosphatase