臺灣博碩士論文加值系統

食道鱗狀細胞癌 (esophageal squamous cell carcinoma ; ESCC) 為世界癌症死亡率第六名，因早期不易診斷且術後預後差而導致高死亡率，且目前並無有效的治療方式。蜂膠具有抗氧化、抗菌、抗發炎、以及抗腫瘤的功能，蜂膠素C為臺灣蜂膠粗萃取物中分離出的活性物質。蜂膠素C具有抗腫瘤活性，但對於食道鱗狀細胞癌的抗癌機制尚未明瞭。本研究主要探討臺灣蜂膠萃取物蜂膠素C對ESCC細胞生長的影響。結果顯示，臺灣蜂膠粗萃物能夠抑制ESCC細胞生長，其中以蜂膠素C對於ESCC細胞生長抑制效果最佳。透過西方墨點法分析，蜂膠素C會抑制細胞週期相關蛋白、表皮生長因子受體 (epidermal growth factor receptor, EGFR) 及下游路徑的轉錄訊息傳遞活化子3 (signal transducer and activator of transcription 3, STAT3) 的表現，且會增加細胞自噬作用及內質網壓力相關蛋白質的表現。此外，蜂膠素C會誘導活性氧化物質 (reactive oxygen species, ROS) 生成。綜合上述，蜂膠素C會透過抑制ESCC細胞EGFR/STAT3訊息傳遞的活化，進而影響Cyclin D1/CDK4 (cyclin-dependent kinase 4) 對於細胞週期的調控，造成細胞週期停滯，同時誘導產生ROS並發生內質網壓力，促使ESCC細胞誘導發生自噬作用，進而達到抑制食道鱗狀細胞癌生長效果。

Esophageal squamous cell carcinoma ( ESCC ) is the sixth most common cause of cancer-related deaths in the world. ESCC is difficult to early detection and poor prognosis, resulting in high mortality rate. Unfortunately, there is no effective therapeutic strategies for ESCC till now. Propolis is a complex mixture of which the main bioactive compounds are polyphenols. Several studies show that propolis has antibacterial, antiviral, antioxidant, anti-inflammatory and anti-tumor activity. Propolin C, a bioactive compound from Taiwanese propolis, has been demonstrated to possess anti-tumor activity. However the anti-cancer mechanisms of propolin C in ESCC are still unclear. The purpose of this study is to investigate the effect of propolin C on the proliferation of ESCC cells. The results showed that propolis inhibited ESCC cell viability, and propolin C has the best inhibition effect. Propolin C decreased the cycle cycle related protein, EGFR/STAT3 signaling pathway, and increase autophagy and ER stress related protein expression. In addition, propolin C increased ROS levels in ESCC cells. According to our results, propolin C inhibited ESCC cell growth by decreasing EGFR/STAT3 signaling, and influenced the regulation of Cyclin D1/CDK4 in cell cycle. In addition, induction autophagy by propolin C through ROS production and ER stress resulted in the proliferation of ESCC cells suppression.

目錄
第一章、文獻回顧 1
一、食道癌 (Esophageal cancer) 1
(一)、概論 1
(二)、食道癌的類別與疾病發展 1
(三)、臨床上常用治療方法 2
(四)、食道癌與癌症標靶治療 3
二、細胞週期 (Cell cycle) 之相關分子機制 4
(一)、週期素家族 (Cyclin) 5
(二)、細胞週期依賴性蛋白激酶 (Cyclin-dependent kinase ; CDK) 5
(三)、細胞週期與食道鱗狀細胞癌 6
三、表皮生長因子受體 (Epidermal growth factor receptor ; EGFR) 6
(一)、表皮生長因子受體活化與下游路徑 6
(二)、表皮生長因子與食道鱗狀細胞癌 7
四、內質網壓力 8
(一)、癌症與內質網壓力 8
五、自噬作用及細胞凋亡 9
(一)、自噬作用 9
(二)、細胞凋亡 10
六、臺灣蜂膠 11
(一)、蜂膠之來源與分類 11
(二)、臺灣蜂膠的特性 11
(三)、臺灣蜂膠的生理功能 12
1. 抗菌活性 12
2. 抗氧化與清除自由基活性 13
3. 癌細胞毒殺活性 13
(四)、臺灣蜂膠與癌症之間的關係 13
第二章、研究設計與目的 15
一、研究目的 15
二、實驗設計 16
三、實驗設計流程圖 17
第三章、材料與方法 18
一、臺灣蜂膠萃取物蜂膠素C 18
二、實驗材料 18
(一)、細胞株 18
(二)、培養基 18
(三)、藥品試劑 19
(四)、抗體 20
三、實驗儀器 22
四、實驗耗材 23
五、常用溶液配置 24
六、實驗方法 27
七、統計分析 32
第四章、實驗結果 33
(一)、臺灣蜂膠Propolis對食道鱗狀細胞癌之細胞存活率影響 33
(二)、蜂膠素Propolins對食道鱗狀細胞癌之細胞存活率影響 33
(三)、蜂膠素C對食道鱗狀細胞癌之細胞週期影響 34
(四)、蜂膠素C對食道鱗狀細胞癌之細胞週期時間點影響 35
(五)、蜂膠素C對食道鱗狀細胞癌之細胞週期相關蛋白質影響 36
(六)、蜂膠素C對食道鱗狀細胞癌之EGFR相關路徑蛋白質影響 36
(七)、蜂膠素C對食道鱗狀細胞癌之細胞凋亡及自噬作用影響 37
(八)、蜂膠素C會誘導食道鱗狀細胞癌細胞發生細胞質空泡化的現象 38
(九)、蜂膠素C對食道鱗狀細胞癌細胞之內質網壓力相關蛋白質影響 38
(十)、蜂膠素C對食道鱗狀細胞癌細胞中ROS水平之影響 38
(十一)、蜂膠素C誘導食道鱗狀細胞癌細胞產生細胞質空泡化與EGFR之間的關係 39
第五章、討論 40
(一)、臺灣蜂膠及萃取物蜂膠素C對食道鱗狀細胞癌細胞生長的影響 40
(二)、臺灣蜂膠萃取物蜂膠素C抑制食道鱗狀細胞癌細胞生長之訊息傳遞路徑之探討 40
1.細胞週期 41
2.EGFR下游訊息傳遞路徑 42
3.細胞凋亡及自噬作用 43
4.ROS生成、內質網壓力與細胞質空泡化現象 44
5.內質網壓力與自噬作用 46
(三)、總結 47
第六章、圖表 48
第七章、參考文獻 67
第八章、附圖 70
第九章、附表 75

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