[Interactions of ovarian carcinoma cells and human peritoneal mesothelial cells involved in matrix metalloproteinases expressions of ovarian carcinoma cells].
OBJECTIVE
To investigate the interactions of ovarian carcinoma cells and human peritoneal mesothelial cells (HPMC) involved in matrix metalloproteinases (MMP) expressions of ovarian carcinoma cells.


METHODS
The conditioned medium (CM) of ovarian carcinoma cell SKOV3 was tested by enzyme-linked immunosorbent assay (ELISA) for transforming growth factor beta1 (TGF-beta1). The impact of SKOV3-CM in the presence or absence of TGF-beta1 neutralizing antibody on fibronectin (Fn) gene expression of HPMC was studied by RT-PCR. HPMC were pretreated with serum-free medium, SKOV3-CM, SKOV3-CM + TGF-beta1 neutralizing antibody, and SKOV3-CM + IgG, then the supernatant was collected as HPMC-CM(1), HPMC-CM(2), HPMC-CM(3) and HPMC-CM(4). SKOV3 were incubated with different HPMC-CM, HPMC-CM(1) + antibody against Fn or HPMC-CM(1) + IgG. MMP-2 and MMP-9 gene mRNA expressions and protein expressions of SKOV3 were detected by RT-PCR and ELISA respectively.


RESULTS
TGF-beta1 in SKOV3-CM was (236 +/- 22) ng/L. Fn gene mRNA expressions of HPMC before and after stimulation by SKOV3-CM were 1.328 +/- 0.025 and 2.643 +/- 0.051, and the latter was higher than the former (P < 0.05). Fn gene mRNA expressions of HPMC stimulated by SKOV3-CM + TGF-beta1 neutralizing antibody was 1.897 +/- 0.035, which was less than that of SKOV3-CM group (P < 0.01). Before SKOV3 were incubated with HPMC-CM, MMP-9 protein and mRNA expressions were (14.5 +/- 1.6) microg/L and 1.50 +/- 0.04, but protein and mRNA expressions of MMP-2 were scarcely detected. When SKOV3 were incubated with HPMC-CM(1), mRNA expressions of MMP-2 and MMP-9 were 0.226 +/- 0.012 and 2.66 +/- 0.07, protein expressions were (15.0 +/- 0.8) and (37.2 +/- 3.5) microg/L, which were all higher than those of SKOV3 without treatment of HPMC-CM (P < 0.01). Following incubation of SKOV3 with HPMC-CM(1) + antibody against Fn, mRNA expressions of MMP-2 and MMP-9 were 0.138 +/- 0.007 and 1.82 +/- 0.06, protein expressions were (8.8 +/- 0.7) and (25.8 +/- 2.5) microg/L, which decreased compared with those of HPMC-CM(1) group (P < 0.01). Following incubation of SKOV3 with HPMC-CM(2), mRNA expressions of MMP-2 and MMP-9 were 0.467 +/- 0.018 and 4.28 +/- 0.09, protein expressions were (39.3 +/- 3.6) and (62.0 +/- 5.3) microg/L, which were higher than those of HPMC-CM(1) group (P < 0.01). Following incubation of SKOV3 with HPMC-CM(3), gene expressions of MMP-2 and MMP-9 were 0.331 +/- 0.015 and 3.52 +/- 0.08, protein expressions were (27.6 +/- 1.9) and (50.0 +/- 4.1) microg/L, which decreased compared with HPMC-CM(2) group (P < 0.05), but were still higher than those of HPMC-CM(1) group (P < 0.05).


CONCLUSIONS
Ovarian carcinoma cells activate HPMC through TGF-beta1, which induces higher expressions of MMP of ovarian carcinoma cells. Up-regulating Fn expression of HPMC may be one of action mechanisms of ovarian tumor cells. Fn derived from HPMC stimulates MMP-2 and MMP-9 expressions of ovarian carcinoma cells at gene and protein levels.
Zhang
Jing-jing
JJ
Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, China.
Wang
Bo
B
D000906:Antibodies / Q000494:pharmacology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000235:genetics; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D010537:Peritoneum / Q000166:cytology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis / Q000276:immunology
D000906:Antibodies; D017077:Culture Media, Conditioned; D005353:Fibronectins; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9