Caveolin-1, Through its Ability to Negatively Regulate TLR4, is a Crucial Determinant of MAPK Activation in LPS-challenged Mammary Epithelial Cells

Abstract

<strong>Background:</strong> To explore the role of caveolin-1(CAV-1) gene silencing on MAPK activation in lipopolysaccharide(LPS)-challenged human mammary epithelial cells. <br /><strong>Methods</strong>: We established a MCF-10ACE of CAV-1 genesilencing from human mammary epithelial cell line MCF-10A by RNAi technology. DNA Microarray wereused to detect the expression of inflammation-associated genes in MCF10ACE. Western blotting was used toexamine the activation of MAPK in lipopolysaccharide(LPS)-challenged MCF-10A and MCF-10ACE. Moreover,immunofluorescence and Western bloting were performed to detect the co-localization of CAV-1 and toll-likereceptor 4 (TLR4) in human mammary epithelial cells. <br /><strong>Results</strong>: MCF-10ACE exhibited significant increases ininflammation-associated gene expression, especially IL-6 (~7-fold) and IL6R (~17-fold). In addition, LPS-inducedp38 MAPK and JNK MAPK activation was significantly increased in MCF-10ACE. Furthermore, CAV-1 colocalizedwith TLR4 and appeared a negative correlation trend. <br /><strong>Conclusion</strong>: CAV-1 gene silencing promotesMAPK activation via TLR4 signaling in human mammary epithelial cells response to LPS.