人類腫瘤易感基因 TSG101，它的功能包括細胞內蛋白質分送、液泡的傳輸、基因轉錄及上皮細胞生長及分化之調控。在 TSG101基因啟動子序列中具有 Sp1的結合位點，但不具有 TATA box，為一典型管家基因，文獻指出，TSG101調控皮膚角化上皮細胞的分化是受到 PKC-Sp1的訊息之引導。本研究在探討不同時期之子宮頸上皮內贅瘤檢體中， TSG101與 Sp1表現的相關性。檢體源自財團法人癌症健康篩檢中心病理科所收集從2005年1月到2007年7月的子宮頸上皮組織檢體的臘塊共129例，包括41例正常及88例子宮頸上皮內贅瘤之檢體（35例 CIN I，28例 CIN II，25例 CIN III/CIS）。檢體以免疫組織化學染色後，結果以 image pro-plus 6.1 軟體進行顯微影像量化分析，再藉由 SPSS 統計分法來分析比較在不同程度病灶的細胞中 TSG101及 Sp1兩種蛋白質表現之相關性。結果發現 TSG101 與 Sp1 在正常的子宮頸上皮細胞主要表現在副基底層及中間層的細胞內，而在基底層及快要老化的表皮層的細胞內，則 TSG101與 Sp1不會表現或呈現弱陽性，但隨著子宮頸上皮細胞異常生長的情況愈嚴重(CIN II、CIN III/CIS)，其 TSG101與 Sp1表現都有顯著增加。此結果與先前報導認為 TSG101在誘導上皮細胞的生長及分化的過程中，是透過 Sp1之訊息傳遞所調控之結果相符合。本研究結果顯示在正常子宮頸上皮細胞中 TSG101與 Sp1蛋白質有一致之表現關係，亦間接確認 Sp1訊息調控 TSG101之表現在上皮細胞生長分化之重要性；此外，在不同時期之子宮頸上皮內贅瘤檢體中 ，TSG101 及 Sp1蛋白之表現可能為重要標幟，而值得進行更多的臨床檢體之分析，以確認其作為預後指標之可能性。

Human tumor susceptibility gene 101, TSG101, exhibits a variety of functions including protein sorting, vesicular trafficking, and regulation of transcription, epithelial growth and differentiation. The upstream sequence of TSG101 gene shows a typical housekeeping TATA-less and Sp1 containing promoter. Our previous data indicated the essential role of TSG101 in skin keratinocyte differentiation that is under the regulation of PKC-Sp1 signaling. In this report, we investigated the correlation of TSG101 and Sp1 expression in the specimens of cervical intraepithelial neoplasia. Cervical intraepithelial neoplasia specimens used in this study were 129 paraffin blocks from 41 normal, 35 CIN I, 28 CIN II and 25 CIN III/CIS patients collected in Cancer Prevention and Screening Center at Kaohsiung from January 2005 to July 2007. The expression of TSG101 and Sp1 were analyzed by immunohistochemistry and digitally quantified by Image Pro-plus 6.1 Microimage software according to the method described by Eliane Pedra Dias et al. The quantified data were statistically analyzed using Spearman's rho coefficient and SPSS for Win, v.14 statistical software (SPSS Inc., Chicago, USA). Values were considered significantly different when the P value < 0.05. We found that TSG101 and Sp1 are expressed in cells of parabasal and intermediate layers in normal cervical epithelium, whereas their expressions in basal and superficial layers were either absence or reduced. Interestingly, the expressions of these two markers are significantly increased in more advanced progression stages (CIN II and CIN III/CIS) of cervical intraepithelial neoplastic specimens (P < 0.05). Congruous expression pattern of TSG 101 and Sp1 in normal cervical epithelium confirms the important of cellular Sp1 signaling in regulating TSG101 expression, which is essential during epithelial cell growth and differentiation. Our results also indicate upregulation of these two markers might be important for the progression of cervical intraepithelial neoplasia. Further analysis using more specimens should reveal the prognostic value of these two markers.

中文摘要--------------------------------------------------------p. 1
英文摘要--------------------------------------------------------p. 3
背景介紹--------------------------------------------------------p. 5
子宮頸上皮內贅瘤----------------------------------------p. 5
人類腫瘤易感基因(TSG101 ) --------------------------p. 8
Specificity protein 1（Sp1）----------------------------p. 12
實驗目的--------------------------------------------------------p. 16
實驗材料與方法-----------------------------------------------p. 16
H&E 染色------------------------------------------------p. 18
免疫組織化學染色--------------------------------------p. 19
影像分析--------------------------------------------------p. 20
統計分析--------------------------------------------------p. 21
實驗結果與討論-----------------------------------------------p. 21
參考文獻--------------------------------------------------------p. 29
附圖--------------------------------------------------------------p. 34

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