在許多種類型的人類癌症中都有發現Epidermal growth factor receptor（EGFR）的突變或過量表達。最近，EGFR被發現會調節腫瘤相關巨噬細胞（TAM）的M1-和M-2-型的極化，並造成肝癌的形成。Epidermal growth factor receptor pathway substrate number 8（Eps8）在癌細胞和巨噬細胞中都扮演很重要的角色。像EGFR一樣，Eps8的表達在許多種癌症中也有升高的現象。此外，Eps8參與巨噬細胞中的LPS-TLR4信號傳導。Protein kinase C（PKC）是一種serine/threonine kinase，可將多種細胞信號傳導與癌症聯繫起來。已經發現，PKCs，特別是PKCδ，在調節巨噬細胞功能中扮演著很重要的作用。在這項研究中，我們想研究在TAM中EGFR-Eps8的途徑是如何被調控的，以及該途徑是否在大腸直腸癌中調控腫瘤的進展。首先，我們利用SW480共培養的THP-1細胞來模擬TAM。我們發現THP-1細胞在經過PMA（一種PKC激活劑）處理後會從單核細胞分化為巨噬細胞。在此過程中，利用PKC抑製劑 Rottlerin，PKCδ siRNA和eps8 siRNA 證明了PKCδ調控Eps8與EGFR的表達以及Eps8參與PKCδ調控EGFR的表現。此外，Src抑製劑可以下調THP-1細胞中EGFR的表達。接下來，我們觀察到經過SW480細胞共培養或與SW480細胞製備的條件培養基一起養時，巨噬細胞中Eps8和EGFR的表達也會提高。我們的研究結果指出PKCδ，Eps8和Src可能介導THP-1細胞中EGFR的表達，而SW480的分泌物也可以誘導這種作用。

Epidermal growth factor receptor (EGFR) mutation or overexpression has been found in several types of human cancer. Recently, EGFR has been implicated in regulating M1- and M-2-type polarization in tumor-associated macrophages (TAMs) and contributes to hepatocellular carcinoma formation. Epidermal growth factor receptor pathway substrate number 8 (Eps8) which plays an important role in cancer cells and macrophages. Like EGFR, Eps8 expression is also elevated in many kinds of cancer. In addition, Eps8 participates in lipopolysaccharide-TLR4 signaling in macrophages. Protein kinase C (PKC) is a serine/threonine kinase which links multiple cell signaling to cancer formation. Among PKCs, PKCδ have been found to play an important role in regulating macrophage function. In this study, we want to investigate how EGFR-Eps8 pathway is regulated in TAMs and whether this pathway regulates tumor progression in colon cancer. In the beginning, we study SW480-co-cultured THP-1 cells to mimic TAMs. First, we found that during THP-1 cells differentiating from monocytes to macrophages by phorbol 12-myristate 13-acetate (a PKC activator) treatment, the expression of EGFR is elevated in PKCδ and Eps8-dependent manner, which was confirmed by PKC inhibitor Rottlerin, PKCδ siRNA and eps8 siRNA. Moreover, Src inhibitor PP2 also could downregulate the expression of EGFR in THP-1 cells. Next, we observed the expression of Eps8 and EGFR in macrophages is elevated by either co-cultured with SW480 cells or incubation with the condition medium prepared from SW480 cells. Importantly, the expression of CD206, a M2-type macrophage marker, also increase in SW480 co-cultured macrophage. Our study indicated that PKCδ, Eps8 and Src might mediate the expression of EGFR in THP-1 cells, and SW480 secretion could induce this effect.

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