【摘要】：目的探討巨噬細(xì)胞表達(dá)分泌的抗菌肽Hcap18/LL37在卵巢癌發(fā)展中的作用及調(diào)控機(jī)制。方法構(gòu)建卵巢癌細(xì)胞株OVCAR-3與巨噬細(xì)胞共培養(yǎng)模型,采用Matrigel小室檢測(cè)巨噬細(xì)胞對(duì)OVCAR-3侵襲力的影響;Western印跡法和qRT-PCR檢測(cè)Hcap18/LL37和Versican V1蛋白和mRNA的表達(dá)水平。使用干擾質(zhì)粒抑制OVCAR-3細(xì)胞中Versican V1的表達(dá),分析巨噬細(xì)胞中Hcap18/LL37表達(dá)和OVCAR-3細(xì)胞的侵襲力。結(jié)果共培養(yǎng)組侵襲穿膜細(xì)胞數(shù)明顯多于OVCAR-3單獨(dú)培養(yǎng)組(P0.05);Hcap18/LL37抗體共培養(yǎng)組侵襲穿膜細(xì)胞數(shù)明顯少于對(duì)照Ig G共培養(yǎng)組(P0.05)。共培養(yǎng)后巨噬細(xì)胞中Hcap18/LL37蛋白和mRNA相對(duì)表達(dá)量高于單獨(dú)培養(yǎng)(P0.01);OVCAR-3細(xì)胞中Hcap18/LL37蛋白和mRNA相對(duì)表達(dá)量與單獨(dú)培養(yǎng)之間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。Versican V1轉(zhuǎn)染卵巢OVCAR-3細(xì)胞與巨噬細(xì)胞共培養(yǎng)24 h后,Versican V1蛋白相對(duì)表達(dá)量高于OVCAR-3細(xì)胞單獨(dú)培養(yǎng)時(shí)(P0.01)。巨噬細(xì)胞與Versican V1沉默OVCAR-3細(xì)胞共培養(yǎng)組侵襲穿膜細(xì)胞數(shù)低于Versican V1高表達(dá)OVCAR-3細(xì)胞共培養(yǎng)組(P0.01)。結(jié)論卵巢癌內(nèi)環(huán)境中巨噬細(xì)胞分泌的抗菌肽Hcap18/LL37促進(jìn)癌細(xì)胞侵襲,其表達(dá)受腫瘤細(xì)胞分泌的Versican V1蛋白調(diào)控。
[Abstract]:Objective to investigate the role and regulatory mechanism of antibacterial peptide Hcap18/LL37 expressed and secreted by macrophages in the development of ovarian cancer. Methods the co-culture model of ovarian cancer cell line OVCAR-3 and macrophage was established. The effect of macrophages on the invasiveness of OVCAR-3 was detected by Matrigel chamber, and the expression of Hcap18/LL37, Versican V1 protein and mRNA was detected by Western and qRT-PCR. The expression of Versican V1 in OVCAR-3 cells was suppressed by interfering plasmid, and the expression of Hcap18/LL37 in macrophages and the invasiveness of OVCAR-3 cells were analyzed. Results the number of invasive transmembrane cells in co-culture group was significantly higher than that in OVCAR-3 alone culture group (P 0.05), and the number of invasive transmembrane cells in Hcap18/LL37 antibody co-culture group was significantly lower than that in control Ig G co-culture group (P 0.05). The relative expression of Hcap18/LL37 protein and mRNA in macrophages after co-culture was higher than that in alone culture (P 0.01). There was no significant difference in the relative expression of Hcap18/LL37 protein and mRNA between OVCAR-3 cells and culture alone (P 0.05). Versican V1 was co-cultured with macrophages for 24 hours). The relative expression of Versican V1 protein was higher than that of OVCAR-3 cells cultured alone (P 0.01). The number of invasive transmembrane cells in the co-culture group of macrophages and Versican V1 silencing OVCAR-3 cells was lower than that in the co-culture group of Versican V1 cells with high expression of OVCAR-3 cells (P 0.01). Conclusion the antibacterial peptide Hcap18/LL37 secreted by macrophages in ovarian cancer environment promotes the invasion of cancer cells, and its expression is regulated by Versican V1 protein secreted by tumor cells.
【作者單位】： 河北醫(yī)科大學(xué)第四醫(yī)院婦產(chǎn)科;河北醫(yī)科大學(xué)第四醫(yī)院生殖醫(yī)學(xué)科;
【基金】：河北省衛(wèi)生和計(jì)劃生育委員會(huì)科研基金項(xiàng)目(No.zl20140305)
【分類號(hào)】：R737.31

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