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目的 了解热休克蛋白 HSP90 β在人胃癌组织及多药耐药细胞系中的表达 .方法 SABC免疫组化法 :70例胃癌组织 ,其中高分化腺癌 1 3例 ,中分化腺癌 32例 ,低分化腺癌1 8例 ,粘液癌 7例 (包括粘液腺癌和印戒细胞癌 ) .正常胃粘膜 1 7例 ,胃炎 2 3例及癌旁组织 35例作为对照 .组织切片用正常羊血清封闭后 ,加入一抗 (山羊抗人 HSP90 β多克隆抗体1∶ 2 0 0 ) ,4℃冰箱过夜 ;加二抗 (生物素化兔抗山羊 Ig G 1∶2 0 0 ) 37℃ 30 min;加 SABC-过氧化物酶复合物 (1∶ 1 0 0 ) ;DAB显色 .m RNA原位杂交方法 :探针变性后 ,加至组织块上 ,37℃杂交 48h;依次用 2× SSC,1× SSC,0 .5× SSC反复振洗 ,加碱性磷酸酶标记的抗 Dig抗体 ,NBT/ BCIP呈色 .结果 HSP90β主要分布于组织细胞的胞质 .在非癌胃粘膜HSP90 β为弱阳性表达 .正常胃粘膜组织、胃炎 (浅表性及萎缩性 )及癌旁组织 HSP90β的阳性表达率分别为 1 1 .8% ,1 3.0 %及 1 1 .4% ,各组之间无显著差异 (P>0 .0 5 ) .在胃癌组织中 HSP90 β的表达明显增强 ,阳性表达率明显增高 ,阳性率为 30 .0 % (P<0 .0 5 ) .其中在高、中、低分化腺癌及粘液腺癌中 HSP90 β的阳性率分别为 1 5 . 4% ,31 . 3% ,33. 3%及42 .9% . HSP90β的表达有随组织分化程度降低而增高?
Objective To understand the expression of heat shock protein HSP90 β in human gastric cancer tissues and multidrug resistant cell lines. Methods SABC immunohistochemistry method: 70 cases of gastric cancer tissues, including 13 well-differentiated adenocarcinomas and 32 moderately-differentiated adenocarcinomas. There were 18 poorly differentiated adenocarcinomas and 7 mucinous carcinomas (including mucinous adenocarcinoma and signet ring cell carcinoma). There were 17 normal gastric mucosa, 23 gastritis, and 35 paracancerous tissues as controls. After blocking, a primary antibody (goat anti-human HSP90 β polyclonal antibody 1:200) was added and kept in a refrigerator at 4°C overnight; secondary antibody (biotinylated rabbit anti-goat Ig G 1∶200) was added at 37°C for 30 min; Add SABC-peroxidase complex (1: 1000); DAB chromogenic. In situ hybridization of m RNA: the probe is denatured, added to the tissue block, and hybridized at 37°C for 48h; followed by 2×SSC, 1× SSC, 0.5×SSC repeated washing, plus alkaline phosphatase labeled anti-Dig antibody, NBT/BCIP color. Results HSP90β mainly distributed in the cytoplasm of tissue cells. In the non-cancerous gastric mucosa HSP90 β is weak Positive expression. The positive expression rate of HSP90β in normal gastric mucosa, gastritis (superficial and atrophic) and adjacent tissues was 11.8%, respectively. , 1 3.0% and 11.4%, there was no significant difference among the groups (P> 0.05). The expression of HSP90β was significantly increased in gastric cancer tissues, the positive expression rate was significantly higher, the positive rate was 30.0. % (P<0.05). Among them, the positive rate of HSP90 β in high, moderate, poorly differentiated adenocarcinoma and mucinous adenocarcinoma were 15.4%, 31.3%, 33.3% and 42. 9%. The expression of HSP90β increased with the degree of tissue differentiation.