3D Matrigel培養SCC-15研究口腔癌惡化之機轉

中山醫學大學機構典藏 CSMUIR > 醫學科技學院 > 醫學檢驗暨生物技術學系暨碩士班 > 博碩士論文 > Item 310902500/10144

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/10144

Title:	3D Matrigel培養SCC-15研究口腔癌惡化之機轉 Matrigel-embedded 3D culture of SCC-15 cells to study the mechanisms of carcinoma progression
Authors:	陳翊民 Chen, Yi-Min
Contributors:	中山醫學大學:醫學檢驗暨生物技術學系;李明偉
Keywords:	口腔鱗狀上皮癌;基質基底膜;3D腫瘤球體 squamous cell carcinoma;Matrigel;3D tumor spheroids
Date:	2014
Issue Date:	2014-12-10T04:11:31Z (UTC)
Abstract:	口腔癌在國人十大癌症死因中排名第五位，在男性死亡排名在第四位，發生率與死亡率逐年增加，且罹患年齡也有年輕化的趨勢。口腔癌中，鱗狀上皮癌占90%以上，好發部位為舌部，其次為頰黏膜、牙齦、顎、口底部及口唇部。本實驗欲研究人體口腔癌的惡化機轉，以口腔鱗狀上皮癌 (squamous cell carcinoma, SCC)做為實驗模型，加入基質基底膜 (BD Matrigel)混合後製作成3D腫瘤球體，並另以2D平面培養比較差異性。首先利用細胞生長速率分析 (WST-1 Cell viability and proliferation assay)比較細胞在2D平面培養及3D腫瘤球體培養的生長差異，並以蘇木精與伊紅染色 (Hematoxylin and Eosin Y stain, H&E stain)觀察兩組別中的細胞生長型態。以掃描式電子顯微鏡 (Scanning electron microscopy, SEM)拍攝細胞在材料中的生長位置；再以反轉錄聚合?鏈鎖反應 (Reverse transcription polymerase chain reaction; RT-PCR)研究p53, MMP2, K-Ras, Erk基因表現量，並觀察在3D組別中口腔癌的惡化與MAPK pathway間的關係。實驗結果確認3D腫瘤球體為立體環境培養，使細胞型態為球狀，並在材料中均勻分布，且具較佳生長速率，在訊息傳遞路徑的表現中也觀察到惡化情況較2D平面培養明顯；顯示細胞在3D腫瘤球體培養的表現較2D平面培養更接近人體真實狀況，提供臨床研究口腔癌細胞除了動物培養外，另一個腫瘤球體培養的實驗平台。 The death rate of people in Taiwan who suffer from oral cancer in fifth of people's top ten causes of death in cancer, and in forth of men. Head and neck squamous cell carcinoma (HNSCC) has a relatively high mortality rate and poor prognosis, and the mortality has a rising trend and the younger age. In oral cancer, squamous cell carcinoma (SCC) accounting more than 90%, most oral cancer commonly involves the tongue. It may also occur on the cheek lining, floor of the mouth, gingiva, palate or lips. Our experiment investigated the mechanism of deterioration in oral cancer. We used the squamous cell to be the experimental model. To compare with 2D cell culture, we added the matrigel to form a 3D tumor spheroids. We used WST-1 cell viability and proliferation assay to compare the different between 2D culture and 3D group. To confirm the cell morphology, hematoxylin and eosin Y stain was be used to stain the samples. Photos from SEM showed the interaction between cell and matrigel. Reverse transcription polymerase chain reaction would consider gene expression of p53, MMP2, K-Ras, Erk and the interrelation between MAPK pathway and the deterioration of oral cancer. In these study we evidenced 3D promote a prior ability to oral cancer cell to proliferation and deterioration.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/10144
Appears in Collections:	[醫學檢驗暨生物技術學系暨碩士班] 博碩士論文

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