胃癌组织芯片快速制备及Reg Ⅳ编码产物检测  

Fast establishment of tissue microarray of gastric cancer and the detection of Reg Ⅳ encoding product

作　　者：陈说[1] 勾文峰[1] 牛哲峰[1] 赵爽[1] 郑鑫[1] 赵杨[2] 高野康雄 郑华川[1] 

机构地区：[1]中国医科大学基础医学院生物化学与分子生物学研究室,辽宁沈阳110001 [2]中国医科大学附属第一医院妇科,辽宁沈阳110001 [3]神奈川县立癌中心临床研究所

出　　处：《中华肿瘤防治杂志》2014年第13期980-984,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172371);沈阳市科技攻关项目(F11-264-1-10;F12-277-1-01);沈阳人才资源开发基金(2009-01);教育部归国人员科研启动基金(2009-03);人事部留学人员科技活动择优资助项目(2008-01)

摘　　要：目的:建立快速胃癌组织芯片(tissue microarray,TMA)制作方法,探讨其在RegⅣ编码产物检测和研究中的应用。方法:收集日本富山大学医学部1993-04-20-2010-11-29胃癌石蜡标本372例和相应癌旁黏膜标本(距离癌组织>4cm)198例,应用AZUMAYA芯片机构建48阵列组织芯片,利用免疫组化和原位杂交检测RegⅣ蛋白和mRNA的原位表达。在Olympus BX41显微镜下选定切取部位,利用2mm的穿刺针切除供体蜡块,不用制备受体蜡块,直接将切除组织放入金属包埋皿的固定盒上制备48阵列芯片。结果:组织芯片经HE染色确定了组织病理学诊断,免疫组化和原位杂交均发现在肠上皮化生和黏液分泌性肿瘤细胞质中存在RegⅣ蛋白和mRNA表达。胃黏膜肠化生中RegⅣ蛋白表达为100%(63/63),高于胃炎的29.0%(27/93,χ2=73.67,P<0.001)、腺瘤的85.7%(36/42,χ2=9.54,P<0.005)和胃癌的45.2%(168/372,χ2=65.06,P<0.001)。胃癌组织中,印戒细胞癌阳性率为95.3%(41/43),高于低分化癌的27.4%(32/117),χ2=55.89,P<0.001。原位杂交发现,癌旁黏膜肠化生上皮细胞中RegⅣmRNA表达阳性率为95.2%(40/42),高于癌旁正常黏膜的38.6%(17/44,χ2=30.63,P<0.001)和胃癌的20.3%(14/69,χ2=55.74,P<0.001)。结论:制备TMA不需要制备受体蜡块,提高了TMA制作速度。异常RegⅣ表达与胃黏膜肠化生-球样异型增生-印戒细胞癌的发生途径关系密切。OBJECTⅣE：This study aimed to establish the method of tissue microarray （tissue microarray,TMA） of gastric cancer and to explore its application in the detection of Reg Ⅳ encoding product and research. METHODS：We collected 372 paraffin-embedded specimens of gastric cancer and 198 adjacent mucosa （over 4 cm from cancer tissue） from the Affiliated Hospital,University of Toyama, between 1993-04 20 and 2010-11-29. We established the 48-core TMA using AZUMAYA tissue microarrayer. Reg Ⅳ protein and mRNA were detected in TMA of gastric cancer and adiacent mucosa by immunohistochemistry （IHC） and in situ hybridization（ISH）. Under Olympus EX41,we marked the preferable portion of cancer slides and punched out the donor blocks using 2mm needle. The small tissues were put into tissue tank without receptor block making and fixed by cassett to establish 48-core TMA. RESULTS： Under Olympus BX41, we marked the oreferable oortion of cancer slides and punched out the donor blocks using 2mm needle. The small tissues were put intotissue tank without receptor block making and fixed by cassett to establish 48-core TMA. Hematoxylimeosin staining was performed on TMA to confirm pathological diagnosis. IHC and ISH staining displayed the positivity of Reg Ⅳ in the cytoplasm of gastric intestinal metaplasia and mucin-producing carcinoma. The expression of Reg Ⅳ protein was higher in in testinal metaplasia （63/63,100%） than that in gastritis （27/93,29. 0%, X2 = 73. 67, P〈0. 001）, adenoma （36/42, 85.7 %, X2 = 9.54, P〈0. 005） or carcinoma （168/372,45.2%, X2 = 65.06, P〈0. 001）. Among gastric cancer, signet ring cell carcinoma showed the higher Reg Ⅳ expression （41/43, 95. 3%） than poorlydifferentiated carcinoma （32/117, 27.4% ,X2= 55.89,P〈0. 001）. ISH showed that the positive rate of Reg Ⅳ mRNA expression （data not shown） was higher in IM （40/42,95.2%） than that in adjacent NNM （17/44,38.6M,X2 = 30.63,P〈0. 001） or carcinoma （14/69, 20.3 %, X2 = 55.74,P〈0. 001�

关 键 词：胃肿瘤 组织芯片 原位杂交 RegⅣ 免疫组织化学 

分 类 号：R735.2[医药卫生—肿瘤]