探討Chitinase-3-like protein 1(YKL-40)在非小型細胞肺癌所扮演之功能角色

Abstract

肺癌是癌症致死率高的癌症之一，且於預防上面臨極大之挑戰。肺癌中非小型細胞 肺癌占全部肺癌百分之八十以上。約五成肺癌病人會癌細胞會轉移至淋巴結、肝、腎上 腺、骨頭及腦。此外近期研究中已有強烈證據顯示癌症發生、轉移及侵犯與發炎反應有 密切之關係，其主因為癌細胞受發炎因子調控所造成之影響，但並非所有發炎因子皆與 癌症發生相關，然而在發炎反應中所分泌之因子影響癌症最引人注意之作用為白血球侵 入、發炎反應所表現之小分子(cytokines and chemokines)、組織重組發生( tissue remodeling)及血管新生(angiogenesis)。多種反應中参與多種調控因子(IL-6, IL-8, TGF- 及 NF-B) 及發生細胞與細胞間之交互作用( 癌細胞與macrophage) 。其中以 Tumor-associated macrophages (TAM)為主要調控發炎與癌細胞之間之關係。先前報告 中指出在非小型細胞肺癌病人組織切片中發現腫瘤組織內含有macrophage，並發現肺癌 細胞與macrophage 相互影響誘發不同cytokine 之分泌，但相互間調控之機制及影響分 子皆未清楚分析。在本實驗室先前研究發現macrophage 培養後之細胞培養液能促使非 小型細胞肺癌的轉移及侵犯，此外，分析此培養液發現5 種主要蛋白質與癌症轉移及侵 犯有相關性，而YKL-40 及為其中之一。然而，YKL-40 在癌症轉移中所扮演功能目前尚 不清楚，因此本計劃主要研究目標為YKL-40 與癌症生成、轉移及侵犯之關係。YKL-40 為分泌型醣蛋白是哺乳類類幾丁質酶蛋白之一，此種類蛋白不具幾丁質酶的催化功能。 然於臨床檢體中發現YKL-40 於各種癌細胞中有高表現量，而於血清中YKL-40 含量濃度 高與癌症病患預後變差有關。而YKL-40 在非小型細胞肺癌所扮演之功能角色尚未被清楚 研究，因此因此本計畫將利用七株不同特性之非小型細胞肺癌細胞株(H2126, CL 1-1, H1437, H23, H838, CL 1-5, and H2009)，深入探討YKL-40 與非小型細胞肺癌之關係。 此外更將研究YKL-40 訊息傳遞機制。最後，將利用臨床樣本進一步探討非小型細胞肺癌 與YKL-40 表現量及腫瘤生長中YKL-40 訊息調控之相關性。而希望未來YKL-40 可能可 做為治療非小型細胞肺癌之標的。在本計畫中將利用in vitro 及in vivo 探討COX Va 與非 小型細胞肺癌轉移之關係。本計畫執行目標如下 1) 分析YKL-40 對非小型細胞肺癌migration and invasion 之關係。(第一年及第二年) 2) 探討YKL-40 影響migration and invasion 之機制。(第二年及第三年) 3) 利用動物實驗模式探討YKL-40 與腫瘤轉移之關係。(第二年及第三年) 4) 病理學分析統計YKL-40 與非小型細胞肺癌相關性。(第一年及第二年)

Lung cancer is the leading cause of cancer death and its prevention is a major worldwide challenge. Non-small cell lung cancer (NSCLC) comprises more than 80% of lung cancers. Approximately one-half of lung cancer patients develop metastases, the most common sites of which are the lymph nodes, liver, adrenal glands, bone, and brain.Increased evidence suggests that chronic inflammation is closely related with cancer development, metastasis and invasion. The possible mechanisms by which inflammation can contribute to carcinogenesis include lymphocytes infiltration, cytokines and chemokines secretion, tissue remodeling and angiogenesis, etc. The inflammatory mediators facilitate the communication between tumor cells and tumor-associated host stromal tissue, thereby accelerating tumor progression. Tumor-associated macrophages (TAM) are key regulators of the link between inflammation and cancer. Previous studies found macrophage in NSCLC specimens by immunohistochemistry staining, and also discovered interplay of cancer cells and macrophages would induce secretion of certain cytokines and chemokines, but the involved mechanism is still obscure. In our recently study, we found the macrophage culture medium could induce NSCLC cell lines migration and invasion. Following, we analysis macrophage culture medium and found that the five major protein are associated with cancer migration and invasion. The one of protein is YKL‐40. However, the function of YKL‐40 is not clear in cancer metastasis. Therefore, we will focus on YKL‐40 correlated with tumor migration and invasion. YKL‐40 , a secreted glycoprotein is a member of mammalian chitinase‐like proteins, but has no chitinase activity. Elevated serum levels of YKL‐40 have been associated with a worse prognosis from a variety of advanced human cancers. Consequently, high serum YKL‐40 levels have been associated with a poor prognosis in patients with several cancer types. However, the role of YKL‐40 has not been established in NSCLC. In this study, we will use characteristics of NSCLC cell lines (H2126, CL1‐1, H1437, H23, H838, CL1‐5, and H2009) as the study model. We will explore YKL‐40 in NSCLC cell lines to detect and compare the phenotypic characteristics; then, we may investigate the transcriptional signaling pathways that are associated with expression of YKL‐40. Finally, we will correlate the expression of YKL‐40 that are associated with YKL‐40 regulatory signaling in tumor progression to the outcome of NSCLC patients. We believe that YKL‐40 may serve as therapeutic targets for NSCLC patients in the future. We attempt to investigate the influence of YKL‐40 on tumor migration and invasion using in vitro and in vivo model system. The specific aims of this study in next three years will be: 1) To investigate YKL‐40 involved in tumor migration and invasion. 2) To study the mechanism of YKL‐40 in tumor migration and invasion. 3) To study YKL‐40 involved in migration and invasion by using mice as an animal model. 4) To correlate YKL‐40 with the outcome of non‐small cell lung cancer patients.