鄭安理2006-07-262018-07-112006-07-262018-07-112003http://ntur.lib.ntu.edu.tw//handle/246246/23623Mucosa-assocaited-lymphoid tissue lymphoma (MALToma) of stomach is the most common extranodal lymphomas of humans. Gastric MALToma is classified into high-grade (HG) and low-grade (LG) subtypes by histological criteria. LG gastric MALToma is characterized by its closely association with Helicobacter pylori (HP) infection; and eradication of HP by antibiotics cures 50-70% of this tumor. However, HG gastric MALToma, in contrast to its LG counterpart, was once believed to consist of highly-transformed cells, of which growth is independent of HP. We were the first group of investigators who demonstrated that nearly 60% of early-stage HG gastric MALToma remains HP-dependent and can be cured by HP eradication (J Clin Oncol 2001; 19:4245-51). Clinically, it is most important to predict HP-independence state of freshly diagnosed HG gastric MALToma, since the latter may progress rapidly if unresponsive to anti-HP therapy. However, histologic or molecular features, which predict HP-dependence of this tumor, remain elusive. We hypothesize that molecular mechanisms of HP-dependence may be different between LG and HG gastric MALToma. For example, genetic aberration such as t(11;18)(q21;q21), which results in chimeric protein API2-MALT1 and is closely associated with HP-independent state of LG gastric MALToma, dose not seem to exist in HG gastric MALToma. This study aims to identify molecular and cellular markers, as well as gene expression profiles, which may help predict HP-dependence state of HG gastric MALToma. Further clarification of the biologic significance and function of novel HP-dependence-relevant genes will be done. In the first year of the project, we have already identified two molecular markers and two immunologic cellular markers that are closely associated with HP-independence state of HG gastric MALToma. We have demonstrated that nuclear expression of BCL10 has a sensitivity of 85.7%, and a specificity of 100% to predict HP-independence of HG gastric MALToma. In addition, confocal immunofluorescence microscopy revealed that nuclear expression of NF-B correlated well with the aberrant BCL-10 nuclear expression; and nuclear expression of NF-B alone predicts HP-independence with a sensitivity of 85.7% and a specificity of 86.4%. We confirmed that t(11;18)(q21;q21) rarely occur in HG gastric MALToma. Therefore, BCL10 nuclear translocation appears to be a major independent event in predicting HP-independence of HG gastric MALToma. We 4 are currently working on clarification of the mechanism and biologic significance of BCL10 nuclear translocation in HP-independent HG gastric MALToma. Two immunologic cellular markers were found to be relevant to HP-independence state of HG gastric MALToma. We showed that non-expression of CD86 of MALToma cells is a useful marker for predicting HP-independence of HG gastric MALToma. The expression of CD86 was detected in 9 (64%) of 14 HP-dependent high-grade gastric MALTomas but in none of 6 HP -independent cases (p=0.010). We reason that loss of co-stimulatory markers may preclude tumor cell/reactive T-cell interaction and contribute to the transition to HP-independent state of HG gastric MALToma. Finally, we discovered that decreased infiltration of NK cells in tumor tissues is associated with HP-independence of HG gastric MALToma. We found that HP -dependent HG gastric MALToma contained significantly higher numbers of CD56 (+) NK cells than HP -independent cases (2.7 ±1.2% versus 0.80±0.70%; p=0.005). CD56 (+) nature killer (NK) cells are thought to suppress the growth of HP- related autoreactive and neoplastic B lymphoid cells of the stomach. These molecular and immunologic cellular markers will be immediately useful for the physicians to select the right modality of treatment for patients with HG gastric MALToma. We have thus far collected 20 freshly frozen tissue samples of HG gastric MALToma, of which the HP-dependence state has already been verified by prospective clinical study. Microarray approaches to identify gene expression profiles of HP-dependent and HP-independent tumors are ongoing. Among 20 freshly frozen tissue samples of HG gastric MALToma, 16 samples containing microdissectable HG and LG MALToma cells are identified. Comparison of the expression profiles of the co-existing LG and HG counterparts of the same gastric MALToma patient is ongoing. These ongoing studies will help clarify whether the two components of MALToma may belong to two different clones, and genetic changes responsible for HG transformation may not be the same as those responsible for HP –independence transformation.胃黏膜相關性淋巴組織 (Mucosa-Associated Lymphoid Tissue)淋巴癌 是人類最常見之淋巴結外惡性淋巴癌。 胃MALT 淋巴癌依其病理特徵分為低 惡性度和高惡性度。 低惡性度胃MALT 淋巴癌已被證實與幽門螺旋桿菌（H. pylori ）高度相關，且臨床上以抗生素有效清除幽門螺旋桿菌（HP ）後，可 治癒 50-70%的病人。 相對於低惡性度胃MALT 淋巴癌，高惡性度胃MALT 淋巴癌一向被認為是由高度轉型 (transformed) 的細胞組成，其生長不需依賴 幽門螺旋桿菌性 (HP-independence)。 我們的研究團隊率先發現，早期的高 惡性度胃MALT 淋巴癌，仍有60%具HP 依賴性，故在HP 清除後可加以治癒 (J Clin Oncol 2001;19:4245-51)。但高惡性度胃MALT 淋巴癌若對HP 根除治 療無效，其淋巴癌可能會迅速惡化。 因此在臨床上，最好能在治療前預知該 腫瘤是否仍具HP 依賴性，以期能做出最好的治療選擇。 然而到目前為止， 可用來預測高惡性度胃MALT 淋巴癌是否仍具HP 依賴性之相關病理或分子 表徵仍未能充份釐清。 我們假設HP 依賴性之機轉在高惡性度胃MALT 淋巴 癌及低惡性度胃MALT 淋巴癌是不同的。譬如基因變化 t(11;18)(q21;q21)(造成API2-MALT1 雜合蛋白質)，常見於HP 非依賴性的低惡性度胃MALT 淋巴 癌，但卻罕見於高惡性度胃MALT 淋巴癌。 本研究目的即是希望能找出相關之分子、 細胞、或基因表現圖譜標記， 能幫助我們預測高惡性度胃MALT 淋巴癌之HP 依賴性，並進一步探討HP 依賴 性相關基因之功能。 在第一年的研究中，我們已找到與高惡性度胃MALT 淋巴癌之HP 依賴性 有關的兩種分子及兩種免疫細胞標記。首先我們證實細胞核內BCL10 表現，在 預測高惡性度胃MALT 淋巴癌之HP 非依賴性方面，其敏感性可達85.7 %， 而特異性則高達百分之百。此外，共軛焦顯微鏡也證實NF-B 常伴隨BCL10 一起在核內表現。 同時，單獨細胞核內NF-B 的表現，也可預測高惡性度胃 MALT 淋巴癌之HP 非依賴性，其敏感性可達85.7 %，而特異性則達 86.4%。 我們的實驗也證實 t(11;18)(q21;q21) 很少發生在高惡性度胃淋巴癌。 因此細 胞核內BCL10 的轉移，似乎是預測高惡性度胃MALT 淋巴癌之HP 非依賴性中 的最重要獨立因子。 目前我們正積極探討BCL10 細胞核轉移，在高惡性度胃 MALT 淋巴癌HP 非依賴性中所扮演之角色、機制及生理意義。 我們也發現與高惡性度胃MALT 淋巴癌之HP 非依賴性相關的兩種免疫細 胞標記。我們發現，MALT 淋巴癌細胞若無CD86 表現，是預測高惡性度胃 MALT 淋巴癌之HP 非依賴性的重要標記。 在十四位HP 依賴性高惡性度胃 MALT 淋巴癌病人中，有九位淋巴癌細胞中偵測到CD86 表現。 而其他六位6 HP 非依賴性高惡性度胃MALT 淋巴癌病人，其淋巴癌細胞中則完全偵測不到 CD86 表現。 因此我們認為HP 非依賴性高惡性度胃MALT 淋巴胃之生長，可 能並不須淋巴癌細胞與T 細胞之共同作用。 最後，我們也證實癌組織上自然 殺手細胞之浸潤減少，的確與HP 非依賴性有關。 我們發現HP 非依賴性之高 惡性度胃MALT 淋巴癌比HP 依賴性之高惡性度胃MALT 淋巴癌，含有較多的 自然殺手細胞 (2.7 ±1.2% versus 0.80±0.70%; p=0.005)。 這些分子及免疫細胞 標記，可即刻幫助鄰床醫師選擇最適當的方式來治療高惡性度胃MALT 淋巴癌 病人。 我們到目前為止已收集到先前參與前瞻性臨床實驗，並確定HP 之依賴性 與否之20 位高惡性度胃MALT 淋巴癌病人新鮮冷凍標本。 我們正著手進行 基因微陣列分析方法，去探討HP 之依賴性與非依賴性可能有關的基因圖譜 。 在這些冷凍標本中，有 16 位病人之標本含有可微截之低惡性度及高惡性度 MALT 淋巴癌。 最近我們也正進行比較分析同一病人之低惡性度及高惡性度 部份的基因圖譜，這些進行中的研究將有助於了解病人之高及低惡性度腫瘤 是否可能分別屬於兩個不同的clones ，或是造成細胞轉型與HP 非依賴性是分 屬不同的基因變異。application/pdf7272697 bytesapplication/pdfzh-TW國立臺灣大學醫學院內科High-grade gastric MALTomaHP-independenceBCL10NF-κBCD86and CD56 (+)NK cells高惡性度胃黏膜相關性淋巴組織淋巴癌幽門螺旋桿菌非依賴性和自然殺手細胞reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/23623/1/913112B002009.pdf