(1)魚針草內酯在大腸癌的微環境中臨床角色及意義(2)從流行病學上看臺灣糖尿病及女性在膽管癌扮演之角色

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Title:	(1)魚針草內酯在大腸癌的微環境中臨床角色及意義(2)從流行病學上看臺灣糖尿病及女性在膽管癌扮演之角色 1.The role of ovatodiolide and its Clinical implication in colon cancer microenvironement 2.The Interative Role of Diabetes Mellitus and Female Gender on the Risk of Cholangiocarcinoma –A Population-Based Nested Case-Control Study
Authors:	黃彥鈞 Huang, Yan-Jiun
Contributors:	轉譯醫學博士學位學程閻雲
Keywords:	魚針草內酯;大腸癌;微環境;糖尿病;膽管癌 ovatodiolide;colon cancer;microenvironment;Diabetes;Cholangiocarcinoma
Date:	2017-06-22
Issue Date:	2021-01-07 09:59:11 (UTC+8)
Abstract:	摘要（I）在眾多癌症中包括大腸癌，其中YAP1的高表現量有助於腫瘤的形成。但是YAP1在大腸癌形成的角色仍未很清楚。在此，我們將證明YAP1的表現量與M2腫瘤相關巨噬細胞的極化及大腸癌幹細胞的產生有關。經由基因沉默(gene-silencing )或植物生化素（ovatodiolide）來負調控YAP1基因，這不只會抑制大腸癌的形成還會防止M2腫瘤相關巨噬細胞的極化。為了模擬腫瘤及其微環境之間的互動關係及效應，我們將人類單核性細胞株（THP-1）與大腸癌細胞株（HCT116 and DLD-1）共同培養在一起。我們應用流式細胞術（flow cytometry）及即時聚合酶鏈鎖反應（q-PCR）來觀察人類單核性細胞株（THP-1）的M2腫瘤相關巨噬細胞極化。YAP1基因的抑制可經由基因抑制技術或魚針草內酯 ( ovatodiolide )。YAP1基因抑制的分子層次效應可經由集落形成測定（colony formation）, 細胞遷移實驗(migration assay) , 未分化癌症次球體實驗 (colon sphere formation assay)來證時實。在此癌研究中合倂使用服樂癌注射劑（5-FU）與魚針草酯 ( ovatodiolide )。我們將利用同種與異種移植小鼠模型系统來研究YAP1對於大腸癌的產生的角色及腫瘤相關巨噬細胞的浸潤的影響。利用生物信息學的方法，YAP1表現量的增加被發現跟大腸癌病患不好的預後有關。我們發現YAP1表現量會在大腸癌幹細胞中及與M2腫瘤相關巨噬細胞共同培養的大腸癌細胞中增加。YAP1的基因沉默(gene-silencing )會同時導致其它主要致癌路徑上的癌基因如Kras, mTOR, β-catenin的表現量的降低，但是更重要的是促進M2腫瘤相關巨噬細胞產生的細胞激素如IL-4和IL-13表現量的降低。M2腫瘤相關巨噬細胞的共同培養經由動物實驗發發現可以提升大腸癌細胞腫瘤起始能力。魚針草酯 ( ovatodiolide )單獨或與服樂癌注射劑（5-FU）合併使用可以經由CT26同種移植小鼠模型系统的動物實驗明顯顯示抑制腫瘤生成的效應及降低腫瘤相關巨噬細胞的浸潤。我們發現YAP1具有雙重角色，其一是抑制腫瘤生成，其二是阻止M2腫瘤相關巨噬細胞的極化及大腸癌幹細胞的產生。魚針草酯 ( ovatodiolide )的治療經由小鼠模型系统的動物實驗與腸癌細胞實驗顯示會壓制YAP1致癌路徑進而抑制大腸腫瘤的形成及M2腫瘤相關巨噬細胞的極化。魚針草酯 ( ovatodiolide )應可被考慮當做輔助性治療藥物與傳統化療藥物合併使用。摘要（II）糖尿病(DM)據文獻分析會增加肝內（ICC）與肝外 (ECC)膽管癌的風險。但是糖尿病在肝外 (ECC)膽管癌低發生率地區其所扮演的角色及其重要性仍不是很清楚。我們運用以人群為基礎，巢式病例對照研究，主要研究在低發生率的但是高比例肝內（ICC）膽管癌的台灣其糖尿病(DM)及其它危險因子在肝內（ICC）與肝外 (ECC)膽管癌所扮演的角色。我們從台灣癌症登記中收集了從2003至2009年6,093例案被診斷為膽管癌的病案 (肝內（ICC）與肝外 (ECC)膽管癌: 4,695; 1,396)，另外也收集了60,906匹配的對照。跟匹配的對照對比起來，肝內（ICC）與肝外 (ECC)膽管癌的病人容易有糖尿病(DM)。在女性族群及有膽管疾病的人中其膽管癌風險的增加跟糖尿病(DM)有些關聯。另外，跟匹配的對照比起來，糖尿病(DM)在切除膽囊後的病人其肝外 (ECC)膽管癌風險的增加並無相關聯。我們的結果強烈支持糖尿病(DM)與肝內（ICC）與肝外 (ECC)膽管癌的病人有正向的關係; 但是這關係在已接受膽囊切除的病人並不明顯。 ABSTRACT (I) Introduction, Aims and objectives An increased expression of Yes-associated protein (YAP1) has been shown to promote tumorigenesis in many cancer types including colon. However, the role of YAP1 in promoting colon tumorigenesis remains unclear. Here, we demonstrate that YAP1 expression is associated with M2 tumor-associated macrophage polarization and the generation of colon cancer stem-like cells. YAP1 down-regulation by gene-silencing or a phytochemical, ovatodiolide, not only suppress colon cancer tumorigenesis but also prevents M2 TAM polarization. Methods Human monocytic cells, THP-1 and colon cancer cell lines, HCT116 and DLD-1 were co-cultured to mimic the interaction between tumor and its microenvironment. M2 polarization of the THP-1 cells were examined using both flow cytometry and q-PCR techniques. The inhibition of YAP1 signaling was achieved by gene-silencing technique or ovatodiolide. The molecular consequences of YAP1 inhibition was demonstrated via colony formation, migration, colon sphere formation assays. 5-FU and ovatodiolide were used in drug combination studies. Xenograft and syngeneic mouse models were used to investigate the role of YAP1 in colon tumorigenesis and TAM infiltration. Results An increased YAP1 expression was found to be associated with a poor prognosis in patients with colon cancer using bioinformatics approach. We showed that an increased YAP1 expression in the colon spheres, and colon cancer cells co-cultured with M2 TAMs. YAP1-silecning led to the concomitant decreased expression of major oncogenic pathways including Kras, mTOR, β-catenin and more importantly M2-promoting IL-4, IL-13 cytokines. TAM co-cultured colon spheres showed a significantly higher tumor-initiating ability in vivo. Ovatodiolide treatment alone and in combination with 5-FU significantly suppressed in vivo tumorigenesis and less TAM infiltration in CT26 syngeneic mouse model. Discussion We have identified the dual function of YAP1 where its suppression not only inhibited tumorigenesis but also prevented the generation of cancer stem-like cells and M2 TAM polarization. Ovatodiolide treatment suppressed YAP1 oncogenic pathways to inhibit colon tumorigenesis and M2 TAM generation both in vitro and in vivo. Ovatodiolide should be considered for its potential for adjuvant therapeutic development. ABSTRACT (II) Introduction, Aims and objectives Diabetes mellitus (DM) has been associated with an increased risk of extrahepatic cholangiocarcinoma (ECC) and intrahepatic cholangiocarcinoma (ICC). However, the role of DM in a population with a lower incidence of ECC remains unclear. We investigated the role of DM and other risk factors for ECC and ICC by conducting a population-based, nested, case–control study in Taiwan, a region with a lower incidence but a higher proportion of ICC. Methods We identified patients who received a diagnosis of cholangiocarcinoma (CC) from the Taiwan Cancer Registry between 2003 and 2009. A total of 6,093 CC cases (ICC: 4,695; ECC: 1,396) and 60,906 matched controls were included. Results Compared with the controls, the patients with ICC and ECC were more likely to have DM, with an adjusted OR of 1.22 [95% confidence interval (CI): 1.07–1.39] and 1.48 (95% CI: 1.18–1.85), respectively. DM was associated with an increased risk of CC in the women and patients without a history of biliary tract diseases. Moreover, compared with the controls, DM was not associated with an increased risk of ECC in the patients who received cholecystectomy. Discussion These findings strongly support the positive association between DM and the increased risk of both ICC and ECC; however, this association was not observed in the patients who received cholecystectomy.
Description:	博士指導教授:閻雲委員:蕭宏昇委員:沈家寧委員:邱仲峰委員:吳駿翃
Data Type:	thesis
Appears in Collections:	[The Ph.D. Program for Translational Medicine] Dissertations/Theses

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