【作者】 刘勇；

【导师】 张阳德；

【作者基本信息】 中南大学 ， 生物医学工程， 2006， 硕士

【摘要】 目的：研究Bcl-2和Fas／FasL在胆囊癌组织的表达及其相关性，并探讨其生物学意义。方法：应用免疫组化法研究33例胆囊癌，19例胆囊腺瘤，3例胆囊上皮不典型增生和20例慢性胆囊炎组织中Bcl-2蛋白和Fas／FasL蛋白的表达水平及特征。检测了胆管癌细胞株QBC939细胞和人肝门部胆管癌组织中FasL的表达；并用TUNEL技术检测了相关淋巴细胞的凋亡。结果：Bcl-2蛋白的阳性率在胆囊癌、胆囊腺瘤和胆囊上皮不典型增生分别为78.8％(26／33)、73.7％(14／19)和100％(3／3)，三者间无显著差异；而Bcl-2蛋白在胆囊癌和胆囊腺瘤的阳性率均显著高于慢性胆囊炎组织的阳性率40％(8／20)(P＜0.05)。FasL蛋白的阳性率在胆囊癌、胆囊腺瘤和胆囊上皮不典型增生分别为81.8％(27／33)、84.2％(16／19)和100％(3／3)，三者间无显著差异；而FasL蛋白在胆囊癌和胆囊腺瘤的阳性率均显著高于慢性胆囊炎组织的阳性率55％(11／20)(P＜0.05)。Bcl-2蛋白和FasL蛋白的阳性率均与其分化程度和Nevin分期无关。Fas蛋白的阳性率在慢性胆囊炎、胆囊腺瘤和胆囊上皮不典型增生分别为80％(16／20)、73.7％(14／19)和100％(3／3)，三者间无显著差异；而Fas蛋白在胆囊癌的阳性率为39.4％(13／33)，显著低于在慢性胆囊炎和胆囊腺瘤的阳性率(P＜0.05)；Fas蛋白在低分化胆囊癌的阳性率为25％(5／20)，显著低于在高-中分化胆囊癌的阳性率61.5％(8／13)(P＜0.05)，但与Nevin分期无关。Bcl-2蛋白和FasL蛋白在胆囊癌的表达呈显著正相关(r_s=0.478，P＜0.005)，Bcl-2蛋白和Fas蛋白在胆囊癌的表达呈显著负相关(r_s=-0.316，P＜0.05)，Fas蛋白和FasL蛋白在胆囊癌的表达无显著相关性。在所观察的手术切除的肝门部胆管癌组织中均发现FasL蛋白的表达，TUNEL染色发现瘤周肿瘤浸润淋巴细胞(TIL)有高水平的细胞死亡；在胆管癌细胞中检测到FasL蛋白的表达。结论：Bcl-2蛋白和Fas／FasL蛋白的表达与胆囊癌的生物学行为相关，且具有显著的相互协同作用。胆管癌可通过FasL诱导活化淋巴细胞的凋亡以逃避免疫监视。更多还原

【Abstract】 Objective: To study the expression of Bcl-2 protein and Fas/FasL protein in gallbladder carcinoma, and their biological behaviors and correlation. And to assess its ability to induce apoptosis and immune escape. Methods: Avidin-biotin-peroxidase complex method was used to study the expression of Bcl-2 and Fas/FasL in 33 cases of gallbladder carcinoma, 19 cases of gallbladder adenoma, 3 cases of gallbladder epithelial dysplasia and 20 cases of chronic cholecystis. The expression of FasL in 48 specimens of human hilar cholangiocacinoma resected in operation and cultured QBC939 cholangiocarcinoma cell line was detected by immunohistochemistry. TUNEL was used to detect apoptotic cells. Results: The positive rates of Bcl-2 in carcinoma, adenoma and dysplasia were 78.8%(26/33), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Bcl-2 in cholecystis was 40%(8/20). The positive rates of Bcl-2 in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05): The positive rates of FasL in carcinoma, adenoma and dysplasia were 81.8%(27/33), 84.2%(16/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of FasL in cholecystis were 55%(11/20). The positive rates of FasL in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05). The expression of Bcl-2 and FasL were not correlated with metastasis and differentiation. The positive rates of Fas in cholecystis, adenoma and dysplasia were 80%(16/20), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Fas in carcinoma were 39.4%(13/33), which was significant lower than in cholecytis and adenoma (p<0.05). And the positive rates of Fas in poorly-differentiated carcinoma were significantly lower than in well-moderated-differentiated carcinoma (p<0.05), but it was not correlated with metastasis. The expression of FasL and Bcl-2 in gallbladder carcinoma took on some extent positive correlation(r_s=0.478, P<0.005), the expression of Fas and Bcl-2 in gallbladder carcinoma took on some extent negative correlation(r_s=-0.316, P<0.05), the expression of Fas and FasL in gallbladder carcinoma were not significant correlation. Prevalent expression of FasL was detected in 48 resected hilarcholangiocarcinomas examined. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL protein was expressed in cholangiocarcinoma cells. Conclusions: The expression of Bcl-2 and Fas/FasL might be correlated with the biological behaviors of gallbladder carcinoma. And there might be some extent coordinated regulation among them. The hilar cholangiocarcinoma eludes immunological surveillance by inducing, via the Fas/ FasL system , the apoptosis of activated lymphocytes.更多还原