在過去的研究中，我們以胃內視鏡採集胃黏膜上皮組織並分離出具胃幹細胞特質之胃上皮細胞株KMU-GI2。而在細胞培養之過程中，觀察到KMU-GI2 細胞出現自發性轉形的現象，因此將其命名為KMU-CSN。由於此株細胞轉形後具有不死化及快速生長的特性，於是對其進行各種細胞特性之探討；包含免疫染色分析細胞標記、AIG之測試及細胞染色體核型的分析。轉形後之KMU-CSN細胞株大部分已呈現單一上皮細胞型態，且有細胞堆疊生長的現象，其CPDL已超過一百，顯見應已呈不死化。而在染色體型在轉形後CPDL 132時，已有第十二對染色體出現變異（命名為KMU-CS12），最後並以裸鼠實驗証實了此細胞之致癌特性（胃腺癌）。接著我們把第十二對染色體正常之KMU-CSN細胞和已產生變異之KMU-CS12細胞以Agilent Human 1A Oligo Microarray來比較其基因表現差異。結果發現在KMU-CS12細胞中HOXA基因（HOXA4、5、7、9、13）表現比KMU-CSN明顯，所以推論HOXA基因的表現可能與胃癌的生成有關係。因此我們針對臨床收集的人類胃癌組織加以分析其HOXA基因之表現，以探討此基因與胃癌之關係。

In the past study, we used a human gastric stem cell clone, KMU-GI2, isolated from endoscopically biopsied gastric mucosa. As we maintained this KMU-GI2 cell clone, we also found a subclone with spontaneous transformation, named KMU-CSN. We found this KMU-CSN cell line showed homogenous epithelial morphology, cell pile-up appearance due to contact insensitivity, enhanced anchorage independence (from 1.7% of KMU-GI2 to 22% of KMU-CSN) and cell immortalization (CPDL >100). The high tumorigenic potential of this KMU-CS12 cell line has been endorsed by the finding of the tumor formation in nude mice. By spectral karyotyping and SNP GeneChip® Mapping 500K Assay, we found chromosomal abnormalities in chromosome 12, which were derived from duplication of 7p15.1~15.3 and 7p22.1~22.3. In normal cells, HOXA genes were found to localize in chromosome 7. In this KMU-CS12 cell line, an increased expression of HOXA4、5、7、9、13 genes was also shown, as compared with KMU-CSN. With these findings, we hypothesized that HOXA genes may play a role in gastric carcinogenesis. Therefore, we can check the prevalence of HOXA genes abnormalities in human gastric cancer patients.

一、目錄 ------------------------------------3
二、中文摘要 -----------------------------------------5
三、英文摘要 -----------------------------------------6
四、前言與文獻回顧 ---------------------------------7
(一)胃癌 ---------------------------------------------8
(二) Homeobox gene (HOX gene) --------------------13
(三) 實驗架構--------------------------------------- 16
五、材料與方法 -------------------------------------19
(一) 胃癌病人檢體的收集 ---------------------------19
(二) 胃癌組織及biopsy之RNA萃取 ----------------19
(三) 反轉錄反應 ------------------------------------21
(四) 聚合酶連鎖反應 -------------------------------22
(五) Quantitative Real-Time PCR ---------------------23
(六) 西方點墨法 ------------------------------------24
(七) 免疫組織化學染色法(IHC) ---------------------28
(八)統計分析 ---------------------------------------30
六、研究結果 -----------------------------------------31
(一) HOXA基因在各個胃部疾病的表現 ---------------31
(二) HOXA7、9、13可能參與胃癌致癌機轉 ----------35
(三) 在不同胃癌分期中HOXA7、9、13的表現--------37
(四) HOXA9蛋白質在胃部疾病組織中的表現 (Western
blotting) ------------------------------------------38
(五) HOXA9蛋白質在胃癌組織中的表現 (IHC) ------41
七、結果與討論 ------------------------------------45
八、參考文獻 ---------------------------------------50

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