胃癌组织中细胞FLICE抑制蛋白基因的表达研究  

Study on expression of FLICE-inhibitory protein in gastric cancer

作　　者：周晓东[1] 于皆平[1] 于红刚[1] 罗和生[1] 吕农华[2] 

机构地区：[1]武汉大学人民医院消化内科,430060 [2]江西医学院第一附属医院消化内科

出　　处：《中华消化杂志》2005年第1期10-14,共5页Chinese Journal of Digestion

基　　金：国家自然科学基金资助项目(30300154)

摘　　要：目的　本研究检测细胞FLICE抑制蛋白 (c-FLIP)基因在胃癌中的表达及其与临床病理特征的关系,初步探讨c-FLIP在胃癌发生、发展中的意义。方法　收集 48例胃癌及相应癌旁正常组织标本,以免疫组化、Western印迹法检测c FLIP蛋白表达;半定量RT PCR法检测c FLIPmRNA表达;原位末端标记法检测胃癌细胞凋亡指数。结果　胃癌及癌旁正常组织均表达c FLIPmRNA,平均相对吸光度值分别为(0. 59±0. 16)和 ( 0. 24±0. 13 ),前者显著高于后者 (P<0. 01 )。免疫组化结果表明c FLIP蛋白在胃癌中表达的阳性率为 100% (48 /48),且 68. 8% (33 /48)呈强阳性表达,而在癌旁正常组织中的阳性率为 75%,且未见强阳性表达,癌组织中的表达水平 ( 6. 93±0. 58 )显著高于癌旁正常组织(3. 19±0. 26,P<0. 01)。较低表达c FLIP蛋白的癌组织其平均凋亡指数 [ (2. 96±0. 15)% ]明显高于高表达c FLIP的癌组织[ (1. 36±0. 11)%,P<0. 01]。另外,在有淋巴结转移组中c FLIPmRNA(0. 64±0. 18)和蛋白(7. 15±0. 63)的表达水平明显高于无淋巴结转移组的c FLIPmRNA( 0. 52±0. 13,P<0. 05)和蛋白水平(6. 69±0. 47,P<0. 01)。Western印迹分析表明,胃癌组织中长型和短型c FLIP蛋白的水平分别为癌旁正常组织的 2. 6倍(P<0. 01)和 2. 8倍 (P<0.Objective To investigate the expression of cellular FLICE/caspase-8 inhibitory protein(c-FLIP) in gastric cancer and its possible implications. Methods Forty-eight gastric cancer specimens and their normal counterparts were obtained. Western blot and immunohistochemistry were used to detect protein expression of c-FLIP. Semi-quantitative RT-PCR and TUNEL were used to measure c-FLIP mRNA levels and tissue apoptotic index, respectively. Results Mean levels of c-FLIP mRNA were significantly higher in gastric can-(cer) tissues than those in matched normal tissues (0.59±0.16 vs 0.24±0.13, P<0.01). Immunohistochemical results revealed that the level of c-FLIP protein was significantly higher in gastric cancer than in matched normal mucosa (6.93±0.58 vs 3.19±0.26, P<0.01). Relatively higher levels of c-FLIP protein in cancerous tissues were strongly associated with reduced apoptotic index [(2.96±0.15) % vs (1.36±0.11) %, P<0.01] which was not found in matched normal tissues. In addition, the level of c-FLIP was significantly higher in cancers with lymph node metastasis than without it at mRNA levels (0.64±0.18 vs 0.52±0.13, P<0.05) and protein levels (7.15±0.63 vs 6.69±0.47, P<0.01), respectively. Western blot analysis demonstrated that thc expression levels of the long form of c-FLIP and the short form of c-FLIP in cancers were 2.6-fold (P<0.01) and 2.8-fold (P<0.01) higher than those in matched normal tissues, respectively. However, no significant differences were found between the expression levels of the two isoforms. Conclusions The expression of c-FLIP is upregulated in gastric cancer and it may be one of the mechanisms by which tumor cells escape from apoptosis during the malignant transformation, and it may play an important role in lymph node metastasis of the tumor.

关 键 词：C-FLIP 表达水平 癌旁正常组织 胃癌组织 抑制蛋白 淋巴结转移 强阳性 RNA 细胞 基因 

分 类 号：R735.2[医药卫生—肿瘤]