全國中小學科展

醫學與健康科學

探討CHI3L1對M1巨噬細胞極化及其功能之影響

癌症多年來高居國人十大死因之首,過去研究顯示慢性發炎與癌症生成密切相關。在發炎反應中巨噬細胞的角色相當關鍵,但在腫瘤微環境下反而幫助腫瘤細胞生長。CHI3L1廣泛表現在腫瘤組織與發炎相關的疾病,在發炎中可能調控不同的免疫相關細胞。因此我們想探討CHI3L1在腫瘤微環境中的功能,研究CHI3L1是否影響巨噬細胞之極化及其他後續功能。結果證實有CHI3L1存在時,巨噬細胞無法正常表現M1巨噬細胞之特徵基因,且其濃度越高,特徵基因表現量就越低。而利用巨噬細胞抗原呈現試驗也發現CHI3L1會降低M1巨噬細胞的抗原呈現能力。綜合以上結果,得知CHI3L1會導致M1巨噬細胞功能缺失。未來將探討CHI3L1如何調控巨噬細胞內的分子機轉,並尋找小分子藥物阻斷CHI3L1的作用,期待對腫瘤合併治療有所幫助。

以中藥活血藥方透過GDNF媒介路徑作為抑制乳癌轉移之新穎策略

乳癌為最常見的婦女癌症,轉移後之乳癌具有極差的預後與較低存活率,直至目前尚未具有效抑制乳癌轉移的藥物,因此研發一個有效抗轉移的藥物是極為迫切的。許多研究指出促發炎因子如GDNF、STAT3、JAK和腫瘤的轉移有密切關聯,因此本研究使用一個含有多味具抗發炎效果的中藥複方CR-1,探討其是否能應用於抗乳癌轉移。在動物實驗結果我們看到CR-1能夠有效的抑制腫瘤的生長,為了進一步研究將此複方之成分,依循中藥水煎與酒煎的概念處理分為:水層 (CR-1-WF) 與酒精層 (CR-1-EF)。經由Cytokine array指出CR-1-WF處理過後的乳癌細胞,GDNF、CCL5、IL8等與轉移高度相關的促發炎因子表現量有明顯下降。在癒傷實驗和transwell migration assay我們發現CR-1-WF能夠顯著抑制乳癌細胞的移動。這些實驗結果皆顯示未來CR-1有可能成為一個抗乳癌轉移的藥物。

上皮細胞黏著分子(EpCAM)促進大腸癌細胞中對於艾瑞莎(Gefitinib)之抗藥性研究 EpCAM enhances Gefitinib-induced drug resistance in colon cancer cells

上皮生長因子受體 (EGFR) 已被確認在人類上皮惡性腫瘤扮演重要角色,因此臨床上開發出許多針對EGFR之大腸癌、肺癌等的小分子標靶藥物,但治療期間所產生的抗藥性仍是一大瓶頸。 過去上皮細胞黏著分子(EpCAM)只被認為是胞間連接分子,現今則在癌幹細胞 (cancer stem cells,CSCs)等領域被研究。然而從文獻與先前實驗室的試驗,可以看見EpCAM促進癌症抗藥性的可能。 本研究發現在大腸癌中,艾瑞莎 (Gefitinib) 會透過轉錄因子FOXO3a促進細胞凋亡,而EpEX會經由抑制FOXO3a所促進的凋亡路徑,導致癌細胞產生Gefitinib抗藥性,且此抗藥性,也與EpICD下游之誘導性多能幹細胞相關基因 ( iPS-related genes )的表現有關,但其分子機制尚不清楚。 本研究以大腸癌與Gefitinib做為癌症與EGFR小分子藥物的模型,找出EpCAM可能造成的抗藥路徑,未來可應用在各類癌症之聯合治療 (Combination therapy) ,以克服癌症治療所產生的EGFR小分子藥物抗藥性。

探討rVP1在肺癌細胞上Integrin路徑及Wnt/β-catenin路徑的作用機制

經基因重組技術純化的口蹄疫病毒鞘蛋白rVP1被證實在癌細胞內會經由Integrin路徑對癌細胞的轉移及凋亡產生影響。本研究所探討的是rVP1對肺癌幹細胞H1299及肺癌細胞A549中Integrin路徑與Wnt/β-catenin路徑的作用機制,目的除了要了解兩路徑在肺癌細胞中的影響,也希望能找出兩路徑的交互作用機制。 本研究發現肺癌幹細胞H1299中β-catenin會因加入rVP1而表現量下降,與肺癌細胞不同;肺癌細胞A549中Wnt/β-catenin路徑上游的膜上受體Frizzled-8因rVP1作用而表現量下降;而不論在肺癌幹細胞H1299及肺癌細胞A549中Grb2都不會因rVP1作用而表現量有所改變;Wnt/β-catenin路徑下游產物MIG-7則會因加入Wnt3aligand而表現量下降。 由實驗結果推測癌症幹細胞本身與癌細胞特性不同,可能使得Wnt/β-catenin路徑產生的效果不同;Grb2的變化量則需要更多實驗求證。Integrin路徑會影響到Wnt/β-catenin路徑的上游生成,兩者確實有相互影響。

小花蔓澤蘭活性成分Dihydromilanolide誘導胃癌細胞凋亡與自噬性死亡

外來入侵種小花蔓澤蘭繁殖速度太快,使台灣本土生環境及多樣性受到破壞。我們研究發現,小花蔓澤蘭葉萃取物會誘導人類胃癌(AGS)細胞毒性;以HPLC分析及分離出小花蔓澤蘭葉萃取物的活性成分Dihydromilanolide (DHK),發現DHK會毒殺胞胃癌、卵巢癌、乳癌與血癌細胞,其中以胃癌(AGS)細胞毒殺性最強。此外,抗氧化劑N-acetylcysteine可減緩小花蔓澤蘭葉萃取物及DHK對胃癌細胞毒殺性,推測是透過活性氧化物(ROS)來毒殺胃癌細胞。我們亦發現,DHK可與抗癌藥物(Doxorubicin、Cisplatin或Paclitaxel)對胃癌細胞產生協同作用。DHK作用胃癌細胞會誘導Caspase-3增加、PARP蛋白裂解、促凋亡Bax增加及抑凋亡Bcl-2減少;產生酸性囊泡(AVOs)、促自噬LC3-II及Beclin-1蛋白增加,且加入自噬抑制劑 3-MA可保護DHK誘導胃癌細胞死亡。推論 DHK可誘發胃癌細胞凋亡(Apoptosis)及自噬性死亡(Autophagy)。總結,小花蔓澤蘭與活性成分Dihydromilanolide (DHK) 具抗胃癌功效且與抗癌藥物產生協同作用,可開發成抗癌的藥品或保健食品。

TCA cycle perturbation induces renal mitochondrial dysfunction and enhanced oxidative stress in diabetic nephropathy-1H NMR-based untargeted metabolomics analysis for potential biomarkers and the effe

本研究藉由基因誘導之第二型糖尿病小鼠(Male B6.BKS-Leprdb mice; db/db mice)做為動物模型,並利用氫質譜儀與代謝體學之平台(1H NMR-based untargeted metabolomics)了解第二型糖尿病所導致的腎病變之疾病進程中的代謝途徑變化,及白胺酸的介入對於疾病進程的影響。研究發現,三羧酸循環及支鍊胺基酸代謝途徑中的代謝物,在基因誘導之第二型糖尿病小鼠尿液中有顯著下降,此發現指出糖尿病腎病變所造成的粒線體能量代謝功能下降。在肝臟組織萃取中,脂化膽固醇(esterified cholesterol)的上升及 β-胡蘿蔔素(β-carotene)、維生素A(vitamin A)及維生素A酸(retinoic acid)的下降顯示肝臟產生脂肪肝及其氧化壓力之上升。白胺酸的介入對於減緩腎臟粒線體受損及降低脂肪肝與氧化壓力並無顯著功效,此現象可歸因於支鍊胺基酸啟動mTORC1代謝途徑。本研究所標定之代謝物可被應用於第二型糖尿病及其所導致之腎病變的生物指標。

Potential Diagnosis of Cancerous Cells Through Utilising Optical Spectroscopy

Cancer is responsible for an estimated 9.6 million deaths in 2018. Deaths from cancer worldwide are projected to reach over 13 million in 2030. Thus, developing a device that has the capability to solve today’s toughest global challenge is crucial by utilizing a simple yet robust approach - “SEEING THE UNSEEABLE” through bold innovation. Although removing cancer is much more effective than either radiation or chemotherapy, when unseen residual cancer cells remain, they could grow back into tumour overtime. The reoccurrence of cancer contributes to a greater risk of death. Hence, launching a system that is able to distinguish between the cancerous cell and normal cell is ultimately essential to make sure no cancer is left behind during surgery. This robust optical system is established with quantitative approach by exploring the integration of an algorithm into the developed software. The end result of this device has the capability to provide users an accurate numerical pH value. The developed system is integrated with the smart IoT gateway capability whereby this powerful analytical device is incorporated with the real-time monitoring, data transformation and data analyzer. Harnessing the power of technology lets us fight cancer better. Each time a pathologist analyzes tissue after operation, it can take up 2 to 3 days because the tissue has to be frozen, thinly sliced, and stained so it can be viewed under the microscope during the process of biopsy. Thus, it is crucial to invent this Surgeons’ VisionMetric device which has an IoT-based microcontroller that is capable of providing real-time numerical value on-site.

Fabrication and Characterization of Biological Electrospinning Nanofiber Scaffold Based on Cellulose Diacetate-Gelatin-Green Tea for Tissue Engineering Applications

Tissue engineering has developed novel therapies such as many types of wound dressings, bio-pads, scaffolds and bandages, in order to reduce the effects of deep and extensive skin wounds. Here, we have produced an electrospun nanofiber scaffold, based on biodegradable materials such as gelatin (as a natural and hydrophilic polymer) and cellulose diacetate (with optimal biodegradability), in order to increase wound healing using nanotechnology. We also used green tea extract for its anti-oxidant and anti-bacterial effect, to improve the biological properties of the scaffold. In the fabrication process, two polymer solutions: 1. Gelatin (with acetic acid solvent) and 2. Cellulose Diacetate (with acetone solvent) mixed with green tea extract, were prepared. Then they were spun using a two-nozzle electrospinner to produce a hybrid nanofiber scaffold. SEM images showed enough finesse and uniformity of the produced scaffold to simulate the extracellular matrix. Further, measuring the contact angle of water droplet and the web surface, indicated optimal hydrophilicity of the nanofiber scaffold, which controls the level of scaffold degradability and cell adhesion. Also, the results of antibacterial tests for two bacterial strains (E. coli and S. aureus) showed the antibacterial characteristics of the extract-containing scaffold. In addition to previous tests, evaluation of fibroblast morphology on the nanofiber scaffolds, indicated appropriate cell adhesion and expansion, that confirms the biocompatibility of this produced scaffold.

探討MafF對LRH-1調控相關代謝路徑的影響

已知 liver receptor homolog-1(LRH-1)掌控著多種生理功能,在肝臟代謝中扮演著相當重要的角色,我們利用酵母菌雙雜交技術(Yeast two-hybrid)找出數種可能跟LRH-1產生交互作用的蛋白質,並從其中選擇v-maf avian musculoaponeurotic fibrosarcoma oncogene homolog F (MafF)作為主要研究對象,探討其與LRH-1之間的作用對代謝造成的影響。本研究中,我們利用免疫沉澱法證實MafF可與LRH-1形成複合體,而MafF並不影響LRH-1蛋白的表現量。隨後以啟動子活性檢測MafF-LRH-1複合體在細胞內之功能,發現MafF能促進由LRH-1所調控的small heterodimer partner (SHP)、glucokinase IV (Gck)及side chain cleavage enzyme (SCC)等代謝相關啟動子之活性。此外我們以GST-pull down發現MafF與LRH-1結合區域位於LRH-1的DNA 鍵結位(DBD)。綜合上述實驗結果,MafF能與LRH-1的DBD鑑結形成複合體,並促進啟動子活性,未來希望能進一步探究MafF對LRH-1所調控下游基因的影響,以了解MafF於肝臟膽酸合成及醣類代謝機制中所扮演的角色。

熊果酸抗胃癌細胞增生與順鉑的協同效應

胃癌的治療方式以手術及化學藥物治療為主。本研究目的是研究中草藥白花蛇舌草的主要成份熊果酸(Ursolic acid, UA)對於胃癌細胞增生的影響。研究使用兩種人類胃癌細胞株MKN45及SCM-1。藉UA處理胃癌細胞之後,由鏡檢可見細胞數目減少,SRB分析法也證明UA會降低細胞的存活率。利用流式細胞儀,證實UA會讓胃癌細胞凋亡。進一步由西方墨點法實驗顯示,UA會誘導促細胞凋亡蛋白Bax及Bak的上升,並且降低抗細胞凋亡蛋白Bcl-xl及Bcl-2的表現。UA也會抑制訊息傳遞途徑p-Stat3/c-Myc/Cyclin D1。實驗也發現UA合併使用化療藥物順鉑(cisplatin)會使胃癌細胞存活率降低。經由周塔氏藥物合併指數及流式細胞儀分析均與協同順鉑能抑制胃癌細胞的增生,西方墨點法也發現pro-caspase 9 被活化,皆與UA會促進順鉑引發凋亡效應有關,這些實驗證實UA是cisplatin治療胃癌的輔劑,這個結果對中草藥白花蛇舌草的抗癌效果提供了具體的實證。