Mechanisms of antimicrobial peptide LL-37 in macrophage-promoted ovarian cancer cell proliferation
- Author:
- No author available
- Journal Title:
- Chinese Journal of Oncology
- Issue:
- 9
- DOI:
- 10.3760/cma.j.issn.0253-3766.2013.09.005
- Key Word:
- LL-37;巨噬细胞;卵巢肿瘤;肿瘤细胞系;细胞增殖;LL-37;Macrophages;Ovarian neoplasms;Cell line,tumor;Cell proliferation
Abstract： Objective The aim of this study was to investigate the role of macrophages in promotion of ovarian tumor cell proliferation mediated by over-expression of antimicrobial peptide LL-37.Methods To co-culture ovarian tumor ceils SKOV3,3AO and HO-8910 with macrophages.The Transwell(R) inserts system was used in the co-culture model.The effect of macrophages promoted ovarian tumor cell proliferation was assessed by BrdU-ELISA and cell number counting.Expressions of mRNA and protein of LL-37 in the macrophages and SKOV3 cells were determined by RT-PCR and Western blot analysis.To observe that LL-37 is responsible for macrophage-promoted ovarian tumor cells growth,LL-37 neutralizing antibody was added to abrogate the LL-37 activation.Results The cell number assay showed that after 4 days coincubation with macrophages in the proportion of 1∶0.5,the number of SKOV3 cells increased from (6.0 ± 0.5) × 104 to (11.8 ± 1.3) × 104,showing a significant difference (P ＜ 0.05).It also showed that the growth of the SKOV3 cells was dependent on the macrophage number (P ＜ 0.05).The number variability of 3AO and HO-8910 cells was as the same as SKOV3 cells upon co-culture with macrophages.As determined by BrdU-ELISA,the resulted proliferation of ovarian tumor cells was similar to the result of cell number counting.RT-PCR and Western blot results showed that the expression of LL-37 mRNA and protein in the macrophages was remarkably enhanced in a time dependent manner upon coincubation with SKOV3 cells,but did not work in SKOV3 cells.BrdU-ELISA assay exhibited that treatment of cells with LL-37 significantly stimulated HO-8910 and 3AO cell proliferation.Addition of LL-37 neutralizing antibody markedly inhibited macrophage-promoted ovarian tumor cell (SKOV3,3AO and HO-8910 cells) proliferation.The OD values of these three cells were decreased from 2.95 ± 0.11 to 1.45 ± 0.04,from 3.39 ± 0.36 to 1.32 ± 0.09 and from 3.93 ±0.17 to 1.68 ± 0.23,respectively (P ＜ 0.05).Conclusions Over-expression and release of LL-37 from macrophages is responsible for proliferation of ovarian tumor cells in co-culture condition.The data presented indicate that LL-37 may be critical for macrophage-induced tumor progression.
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