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研究生:謝茂志
論文名稱:E-cadherin在胃癌經過熱休克/溫熱療法之後的改變、作用機轉的探討以及臨床意義的研究
論文名稱(外文):Study on the Alteration of E-cadherin in Gastric Cancers after Heat Shock/Hyperthermic Treatment, its Underlying Mechanism and Clinical Significance
指導教授:余家利余家利引用關係
指導教授(外文):Chia-Li Yu
學位類別:博士
校院名稱:國立陽明大學
系所名稱:臨床醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:99
中文關鍵詞:胃癌E-cadherin熱休克溫熱療法
外文關鍵詞:gastric cancerE-cadherinheat shockhyperthermia
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E-cadherin是一種細胞膜上的醣蛋白,分子量約為120 kDa。它會受到蛋白水解作用而斷裂產生分子量約為80 kDa的可溶性蛋白進入血液循環中。E-cadherin在細胞表面介仲同類細胞之間的粘著,卻與癌症的轉移息息相關。當E-cadherin的表現減弱甚或缺乏時,癌細胞容易由原發病灶游離而轉移至他處。利用培養的胃癌細胞株,加以熱休克處理或發炎性細胞激素如IL-1b及TNF-a作用之後,以流式細胞儀、西方墨點法或densitometry定量檢測,都可發現細胞表面的粘著分子,包括CD29,CD54及E-cadherin,皆有表現增強的現象。此結果顯示熱休克處理後,可能藉著E-cadherin的表現增強而抑制癌細胞的轉移。
我們的研究結果顯示胃癌病患的手術前血液中可溶性E-cadherin的濃度(2555 ± 413 ng/ml)高於一般正常人(1432 ± 426 ng/ml,p<0.001)。E-cadherin的濃度高低與胃癌的各種臨床病理因素並無絕對的相關性,但與手術後的存活期有關。我們發現當E-cadherin的濃度高於2563 ng/ml的胃癌患者,五年存活率只有36.4%,而低於2563 ng/ml者,則有83.3%(p=0.0148)的存活。即是血液中可溶性E-cadherin的濃度較低者,預後越好。
在已發生有腹腔內轉移的胃癌病患,在手術後的血液中可溶性E-cadherin濃度會較手術前為低(術前2729 ± 234 ng/ml,術後2437 ± 333 ng/ml,p=0.008)。我們以自行設計的腹腔內溫熱療法(熱休克)裝置,應用於此類病患;即在手術後立刻施行腹腔內溫熱療法,發現術前血液中可溶性E-cadherin的濃度為2674 ± 305 ng/ml,而術後降低為1888 ± 406 ng/ml,p<0.001。只接受手術的病患,術後血液中可溶性E-cadherin的濃度降低了292 ± 226 ng/ml,而手術後再接受腹腔內溫熱療法的病患,則E-cadherin的濃度大幅降低789 ± 388 ng/ml(p<0.001)。
以手術後的存活時間而言,只接受手術的這類病患,平均存活期為8.74個月(1-16個月),而五年存活期為零。手術後再接受腹腔內溫熱療法的病患,平均存活期為31.52個月(1-83個月),而五年存活期為23.81%。我們也發現接受腹腔內溫熱療法的病患中,術後血液中可溶性E-cadherin的濃度若高於1500 ng/ml,則平均存活期為14.67個月(1-39個月),但五年存活期為零。但若血液中E-cadherin的濃度低於1500 ng/ml,則平均存活期為73.67個月(27-83個月),而五年存活期為83.33%。
熱休克會增強胃癌細胞表面E-cadherin的表現,而腹腔內溫熱療法會降低病患血液中可溶性E-cadherin的濃度,兩者都是對於癌症有較佳預後的因素。我們的結論是:熱休克(腹腔內溫熱療法)可能藉著改變E-cadherin的表現提高腹腔內癌轉移病患相當程度的抗癌效果。

E-cadherin, a transmembrane glycoprotein with a molecular weight of 120 kDa, mediates homotypic and homophilic cell-cell adhesion. The breakdown product of E-cadherin, an 80 kDa soluble form of E-cadherin, enters circulation through proteolysis. When membranous E-cadherin expression is down regulated or absent on malignant cells, these cells may escape from the primary lesion and proceed to invade and metastate to the remote sites. In this study, we treated different gastric cancer cell lines with heat shock (42.5°C for 60 minutes)or proinflammatory cytokines (IL-1b and TNF-a). We found the expression of adhesion molecules such as CD29, CD54 and E-cadherin were enhanced on the surface of gastric cancer cells. This result indicates that heat shock has a potential to prevent the cancer from metastasis by alternation in E-cadherin expression on the cell surface.
In addition, we noted that, the serum concentration of preoperative soluble E-cadherin (2555 ± 413 ng/ml) in patients with advanced gastric cancer is higher than that of normal control (1432 ± 426 ng/ml, p<0.001). Although the levels of serum soluble E-cadherin were not correlated to any clinicopathological factors of gastric cancer, it really correlated with the postoperative survival. The 5-year survival rate in patients with preoperative serum soluble E-cadherin levels higher than 2563 ng/ml was 36.4%, whereas the postoperative survival rate increased to 83.3% in those patients with serum soluble E-cadherin levels lower than 2563 ng/ml. The difference in survival rate is significant between the two groups (p=0.0148). These results suggest that the lower serum soluble E-cadherin presents postoperatively, the better the 5-year survival is.
We selected a group of gastric cancer patients with peritoneal carcinomatosis for clinical effectiveness evaluation of hyperthermia therapy. We demonstrated that the postoperative levels of serum soluble E-cadherin are lower than the preoperative levels (2729 ± 234 ng/ml versus 2437 ± 333 ng/ml, p=0.008). The intraperitoneal hyperthermic treatment of the patients right after surgery reduced the postoperative serum soluble E-cadherin to a concentration of 1888 ± 406 ng/ml (preoperation: 2674 ± 305 ng/ml, p<0.001). The reduction of serum soluble E-cadherin in the patients treated with surgery alone was 292 ± 226 ng/ml, while the patients were treated with surgery followed by intraperitoneal hyperthermia was 789 ± 388 ng/ml (p<0.001).
In the patients receiving surgery alone, the mean survival time was 8.74 months (1-16 months) and the 5-year survival rate was zero. However, combining surgery with intraperitoneal hyperthermic treatment (combination therapy), prolonged the mean survival time to 31.52 months (1-83 months) and the 5-year survival rate to 23.81%. For comparison, we further divided the patients receiving combination therapy into two subgroups. (I) The postoperative serum soluble E-cadherin concentration was higher than 1500 ng/ml, the mean survival time was 14.67 months (1-39 months) and the 5-year survival rate was zero. (II) The postoperative serum soluble E-cadherin concentration was lower than 1500 ng/ml, the mean survival time was 73.67 months (27-83 months) and the 5-year survival rate was 83.33%.
In conclusion, heat shock increases the expression of E-cadherin on the surface of gastric cancer cells. The intraperitoneal hyperthermic treatment can reduce the postoperative concentration of serum soluble E-cadherin in these patients. Both alterations in E-cadherin expression are beneficial to the survival of patients with gastric cancer. The molecular basis and clinical implications of heat shock / intraperitoneal hyperthermic treatment are extendedly investigated in the present study. It is conceivable that the anti-cancer effects of heat shock might be related to, at least in part, the modulation of E-cadherin expression.

封面
目錄
致謝
中文摘要
英文摘要
第一章 緣起
1.1 胃癌的治療現況
1.2 癌症的轉移機轉
1.3 E-cadherin的生物活性
第二章 胃癌細胞表面粘著分子經熱休克處理後的改變
2.1 前言
2.2 材料與方法
2.2.1 各種不同來源的胃癌細胞株及其培養
2.2.2 抗細胞表面粘著分子的單株抗體
2.2.3 細胞的熱休克處理
2.2.4 以流式細胞儀來檢測細胞表面粘著分子的存在
2.3 結果
2.3.1 胃癌細胞株表面自然表現的粘著分子
2.3.2 熱休克處理後胃癌細胞株表面粘著分子表現的改變
第三章 胃癌細胞表面粘著分子經細胞激素(cytokines)處理後的改變
3.1 前言
3.2 材料與方法
3.2.1 胃癌細胞株及其培養
3.2.2 使用的細胞激素
3.2.3 細胞的處理
3.2.4 以流式細胞儀檢測細胞表面粘著分子的表現
3.2.5 細胞溶解液的製備及西方墨點分析
3.2.6 胃癌細胞與單核性細胞的粘著反應
3.3 結果
3.3.1 發炎性細胞激素的處理對胃癌細胞株表面粘著分子表現的影響:流式細胞儀分析
3.3.2 發炎性細胞激素的處理對胃癌細胞株表面粘著分子表現的影響:西方墨點分析法
3.3.3 胃癌細胞經熱休克處理對細胞激素產生的影響
3.3.4 胃癌細胞與單核性細胞之間的粘著反應
第四章 胃癌細胞株熱休克處理後可溶性E-cadherin表現的改變
4.1 前言
4.1.1 細胞膜上E-cadherin與可溶性E-cadherin間之關係
4.2 材料與方法
4.2.1 胃癌細胞株及熱休克處理
4.2.2 可溶性E-cadherin之免疫酵素分析法
4.2.3 統計及分析
4.3 結果
第五章 胃癌病患血清中可溶性E-cadherin的濃度及其臨床意義
5.1 前言
5.1.1 利用胃癌病患之癌組織上E-cadherin的表現來監測病患臨床變化的困難點
5.1.2 胃癌病患之癌組織上E-cadherin與血清中可溶性E-cadherin間之關係
5.2 材料與方法
5.2.1 胃癌病患的選擇
5.2.2 血清中可溶性E-cadherin之免疫酵素分析法
5.2.3 統計及分析
5.3 結果
5.3.1 胃癌病患及正常人血清中可溶性E-cadherin的比較
5.3.2 胃癌病患血清中可溶性E-cadherin濃度與胃癌腫瘤病理的關係
5.3.3 胃癌病患血清中可溶性E-cadherin濃度與手術後的存活關係
第六章 胃癌病患接受腹腔內溫熱療法(熱休克)後血清中可溶性E-cadherin的改變
6.1 前言
6.2 材料與方法
6.2.1 腹腔內溫熱療法(熱休克)裝置模式的設計
6.2.2 腹腔內溫熱療法(熱休克)的實際操作
6.2.3 施行腹腔內溫熱療法(熱休克)的胃癌病患的臨床資料
6.2.4 血清中可溶性E-cadherin之免疫酵素分析法
6.2.5 統計及分析
6.3 結果
6.3.1 施行腹腔內溫熱療法(熱休克)的胃癌病患血清中可溶性E-cadherin
6.3.2 血清中可溶性E-cadherin與術後的存活關係
第七章 討論
參考資料
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