IL-13Effect On IKK/NFκBp65Expression Of Human Breast Fibroblasts In The Breast Tumor Microenvironment

ObjectiveBreast has been arranged in the first one of female cancer incidence. Malignanttransformation of breast microenvironment plays an important role in the breasttumor development. Fibroblasts are the most important stromal cells in the breasttumor microenvironment. They interact with each component of themicroenvironment through direct contacting cells, autophagy and secretion of theinformation molecules. They play the role of fertile soil and accumulators in thegrowth of cancer cells, then affecting the occurrence and development of tumors. Theactivity regulatory network of tumor-associated fibroblasts in malignanttransformation microenvironment is a problem being not ignored in cancer relatedfields. In this study, we explore IL-13effect on IKK/NFκBp65expression of thehuman breast fibroblasts in breast tumor microenvironment in order to providetheoretical and experimental basis for the further study of breast tumorigenesis anddevelopment.MethodsHuman breast cancer cells and fibroblasts were co-cultured to set up tumormicroenvironment model in vitro. IKK and NFκBp65mRNA expressions offibroblasts were detected using real-time-PCR technology. IKK and NFκBp65proteinexpressions of fibroblasts were analyzed by cell immunocytochemistry.Results1. Different concentrations of IL-13(5ng/ml,10ng/ml,20ng/ml,100ng/ml)stimulated human breast fibroblasts Hs578Bst co-cultured with human breast cancerMDA-MB-435cells. IKK and NFκBp65mRNA expressions of the co-culturedfibroblasts were detected using real-time-PCR. IKK mRNA expression was nosignificant difference in IL-13100ng/ml group as compared with control (P>0.05).IKK mRNA expression level was significantly up-regulated in IL-13(5ng/ml,10ng/ml,20ng/ml) each group as compared with control(P<0.05). Significantlyincreased expression of NFκBp65mRNA was observed in IL-13(5ng/ml,10ng/ml, 20ng/ml,100ng/ml) each group as compared with control(P <0.05).2. Different concentrations of IL-13(5ng/ml,10ng/ml,20ng/ml,100ng/ml)stimulated human breast fibroblasts Hs578Bst non-co-cultured. IKK and NFκBp65mRNA expressions of the non-co-cultured fibroblasts were detected usingreal-time-PCR. IKK mRNA expression in each experimental group wasdown-regulated as compared with control(P<0.05). NFκBp65mRNA expression,however, was up-regulated in each experimental group as compared withcontrol(P<0.05).3. Under the stimulation of IL-13(5ng/ml,10ng/ml,20ng/ml,100ng/ml), IKKmRNA expression of the co-cultured fibroblasts was higher than the non-co-culturedfibroblasts treated with the same concentration of IL-13(P<0.05). NFκBp65mRNAexpression of the co-cultured fibroblasts in the group of IL-13concentration of10ng/ml was higher than the non-co-cultured fibroblasts in the group of the sameconcentration of IL-13(P<0.05). NFκBp65mRNA expressions of the co-culturedfibroblasts in the groups (IL-13concentrations of5ng/ml,20ng/ml,100ng/ml) werelower than the non-co-cultured fibroblasts in the groups of the same concentration ofIL-13（P<0.05).4. The immunocytochemical results show that IKK and NFκBp65proteinexpressions of the co-cultured fibroblasts is higher than the non-co-culture group (P<0.05).Conclusions1. The joint action of breast tumor microenvironment and IL-13can increase theIKK expression of human breast fibroblasts. IKK expression of human breastfibroblast line Hs578Bs co-cultured with human breast cancer line MDA-MB-435ishigher than the non-co-cultured human breast fibroblast line Hs578Bs.2. IL-13concentration of10ng/ml forms a better synergy with breast tumormicroenvironment, up-regulating NFκBp65expression of human breast fibroblastline Hs578Bs co-cultured with human breast cancer cell line MDA-MB-435.