巨噬细胞通过分泌抗菌肽LL-37/hCAP-18调控卵巢癌细胞的增殖

Proliferation Regulation of Macrophages on Ovarian Cancer Cells by the Secretion of Antimicrobial Peptide LL-37 / hCAP-18

目的研究巨噬细胞是否通过分泌抗菌肽LL-37/h CAP-18对卵巢癌细胞的生长增殖进行调控。方法采用Millicell插入式细胞培养皿共培养人巨噬细胞和卵巢癌细胞株SKOV3;通过细胞计数法检测巨噬细胞对卵巢癌细胞增殖能力的影响;流式细胞术检测共培养上清中的相关细胞因子及LL-37的水平;实时定量反转录聚合酶链反应在mRNA水平检测巨噬细胞和SKOV3细胞中LL-37 mRNA的表达水平;应用LL-37中和抗体抑制LL-37的功能及活性,进而通过MMT细胞增殖实验探讨LL-37对巨噬细胞对卵巢癌细胞增殖的调控。结果细胞计数法检测结果显示,巨噬细胞与卵巢癌细胞SKOV3共培养后,可促进其增殖;共培养的上清中,LL-37及IL-6水平明显升高;实时定量RT-PCR在mRNA水平共培养中的巨噬细胞其LL-37基因表达升高,且其表达水平随时间的延长而增加;应用LL-37的中和抗体,可明显抑制巨噬细胞对SKOV3细胞的增殖促进作用。结论在与卵巢癌细胞共培养的环境中,巨噬细胞通过增强抗菌肽LL-37的表达和分泌水平促进卵巢癌细胞的增殖,LL-37在巨噬细胞促进卵巢癌的发展中扮演重要角色。

Objective To investigate the proliferation regulation role of antimicrobial peptide LL - 37/hCAP - 18 secreted by macrophages on ovarian cancer cells. Methods Millicell system was applied to develop a co - culture system for human macrophages and ovarian cancer cell line SKOV3A. The proliferation effect being exerted by macrophages on ovarian cancer cells was detected by cell counting method. The related cytokines and LL- 37 from the co- cultured supernatants were analyzed byflow cytometry. Real -time quantitative reverse transcription polymerase chain reaction was applied to detect the mRNA level of LL - 37 from macrophages and SKOV3 cells. By applying the LL - 37 antibody to neutralize thefunction and activity of LL - 37, the proliferation role of macrophages on ovarian cancer cell was then further verified by the MMT cell proliferation assay. Results Cell counting test results showed that macrophages and SKOV3 ceils co- cuhured can promote the proliferation. In co -culture supernatant, LL- 37 and IL- 5 levels were significantly increased. For real -time quantitative RT -PCR in mRNA levels of co -culture of macrophages, LL -37 gene expression was increased, and the expression levels increased with time. Application of LL - 37 neutralizing antibody can inhibit macrophage ceils and promote proliferation of SK- OV3 effect. Conclusion Under the co - culture environment, macrophages could promote the proliferation of ovarian cancer cells by enhancing the expression and secretion of antimicrobial peptide LL - 37, therefore, LL - 37 mayplay an vital role in macrophage - induced tumor progression.