Multiple myeloma is characterised by deregulated cytokine network with secretion of inflammatory cytokines. Recent studies have showed the independent poor prognostic value of COX-2 expression in myeloma. We have here studied the COX-2 expression in plasma cells from patients with myeloma, monoclonal gammopathy of undetermined significance (MGUS), Waldenstrom’s Macroglobulinemia (WM) and compared with the COX-2 expression by normal plasma cells.

Methods:

Our study included bone marrow samples from 34 patients (Lymphoma: 15; myeloma: 10; MGUS: 7; WM: 1; Amyloidosis: 1). Mononuclear cells were harvested from the bone marrow samples by density gradient sedimentation using Lymphoprep TM from Axis-Shield, Norway. Mononuclear cells were stained with PE conjugated CD 38 (BD Bioscience) and APC conjugated CD138 (BD Bioscience). FITC conjugated IgG against human COX-2 (Cayman Chemical) was used to study the expression of cycloxygenase-2 following permeabilization with fix and perm kit (Caltag). Flow cytometry was performed on a Becton Dickinson FACS vantage with appropriate isotype controls. The colorectal cell line HT- 29 and human myeloma cell line RPMI 8226 were used as positive controls and the flow results are validated by western blotting and Prostaglandin E2 EIA assay (Cayman Chemical).

Results:

HT-29 cell line showed a peak fluorescence of 58 for COX-2, where as RPMI 8226 cell line revealed peak fluorescence of 141. Median plasma cell count in bone marrow of patients with MGUS was 2 %( range 2–6%). Plasma cells from MGUS patients revealed median peak fluorescence for COX-2 was 37(range: 9–74). Median plasma cell count in bone marrow of patients with myeloma was 25% (range: 10–59%). Median peak fluorescence for COX-2 in plasma cells from patients with myeloma was 24(range: 10–85). Median plasma cell count in bone marrow of patients with lymphoma was 1 %( range1–4%). Median peak fluorescence for COX-2 in plasma cells from patients with lymphoma was 11(range: 2–38).

Conclusions:

Our study reveals COX-2 expression is more in clonal plasma cells as compared to normal plasma cells. Studies are needed to ascertain the role of COX-2 in oncogenesis in myeloma and to discover how COX-2 expression leads to poor outcome in patients with myeloma. This information may lead to the therapeutic role of selective COX-2 inhibitors in the therapy of myeloma.

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