MMP production in human fibrosarcoma cells and their invasiveness are regulated by group IB secretory phospholipase A2 receptor-mediated activation of cytosolic phospholipase A2.

Matrix metalloproteinases (MMPs), especially MMP-2 and MMP-9, are expressed in most colonic, gastric, and ovarian carcinomas, and they play a key role in their invasiveness. Previous studies have shown the involvement of arachidonic acid (AA)-derived metabolites in the regulation of MMP expression and cancer dissemination, thus suggesting a role for phospholipase A2, the AA producing enzymes, in these processes. The present study was undertaken to explore the role of phospholipases in MMP production and tumor cell invasiveness. Human fibrosarcoma cells were found to express and secrete type IB, IIA and V sPLA2. The cells were found also to express the M-type sPLA2 receptor. Treatment with an extracellular sPLA2 inhibitor inhibited tumor cell's invasiveness concomitantly with MMP-2/9 production. Correspondingly, adding an exogenous sPLA2-IB (but not IIA) resulted in significant elevation of MMP-2/9 secretion from the fibrosarcoma cells. Time-course determination of AA and oleic acid release by HT-1080 cells suggested that cPLA2 is activated subsequently to sPLA2 action. Accordingly, using Western blot analysis it was found that sPLA2-IB induced cPLA2 phosphorylation, a requirement for its activation, by a receptor-mediated activity, rather than its lipolytic activity. At the same time, sPLA2-IIA did not induce either MMPs secretion or cPLA2 phosphorylation. The results of this study show for the first time that MMP-2/9 production by human fibrosarcoma HT-1080 cells and their invasiveness is regulated by sPLA2-IB acting as a receptor ligand to activate cPLA2, which in turn provides the AA for production of eicosanoids required for MMP expression.

The clinical role of phospholipase A2 isoforms in advanced-stage ovarian carcinoma.

OBJECTIVE: To analyze the expression of phospholipase A2 (PLA2) isoforms and its relationship with matrix metalloproteinase (MMP) expression and clinical parameters in advanced-stage (FIGO III-IV) ovarian carcinoma. METHODS: Seventy-seven fresh frozen effusions from ovarian carcinoma patients were studied for messenger RNA (mRNA) expression of 10 secretory PLA2 (sPLA2) isoforms (IB, IIA/D/E/F, III, V, X, XII and XIII), the PLA2 receptor (sPLA2R), cytoplasmic PLA2 (cPLA2), PLA2-activating protein (PLAP) and MMP-2 using reverse transcription polymerase chain reaction (RT-PCR). Phosphorylated cPLA2 (p-cPLA2) protein expression was studied in 52 effusions using immunohistochemistry. MMP-2 and MMP-9 activity was evaluated in 22 and 20 effusions, respectively, using zymography. Expression was analyzed for correlation with clinicopathologic parameters, chemotherapy status and survival. RESULTS: PLA2 isoforms, sPLA2R, PLAP and MMP-2 mRNA was expressed in >95% of specimens. p-cPLA2 protein was expressed in 46/52 (88%) effusions. MMP-2 activity was found in all specimens, while that of MMP-9 was detected in 19/20 effusions. MMP-2 was found to be co-expressed with p-cPLA2 (p=0.003) and sPLA2-IIA (p=0.021). Lower expression of sPLA2-IIA (p<0.001) and higher expression of sPLA2-V (p=0.038) and sPLA2-XIII (p=0.001) was found in post-chemotherapy effusions. In univariate survival analysis, higher levels of sPLA2-V correlated with better overall (OS, p=0.021) and progression-free (PFS, p=0.025) survival. For patients with post-chemotherapy effusions, FIGO stage IV and higher PLAP mRNA expression correlated with worse OS (p=0.005 for both PLAP and stage), while higher PLAP (p=0.025) and sPLA2-XII (p=0.027) levels and FIGO stage IV (p<0.001) correlated with shorter PFS. In Cox multivariate analysis, PLAP expression (p=0.022) and FIGO stage (p=0.036) independently predicted poor OS, while higher sPLA2-XII levels (p=0.04) and FIGO stage (p=0.003) were independent predictors of shorter PFS. CONCLUSIONS: The present study documents for the first time expression of PLA2 isoforms, sPLA2R and PLAP in ovarian carcinoma. PLA2 isoenzyme expression differs in pre- and post-chemotherapy specimens. PLAP and sPLA2-XII may be independent predictors of poor outcome for patients with post-chemotherapy effusions.