苏木精染色对冰冻切片组织RNA及蛋白质浓度和质量的影响  

Effect of Hematoxylin Staining on Concentrations and Qualities of RNA and Protein in Frozen Tissue Section

作　　者：王秋悦[1] 张西臣[1] 宫鹏涛[1] 李建华[1] 杨举[1] 曹利利[1] 蔡亚南[1] 续哲莉[2] Yan-gao Man 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]吉林大学中日联谊医院乳腺甲状腺外科,长春130033 [3]Armed Forces Institute of Pathology and American Registry of Pathology, Washington, DC, USA.

出　　处：《中国生物制品学杂志》2008年第11期977-980,共4页Chinese Journal of Biologicals

基　　金：国家重大基础研究计划(2006CB910505)

摘　　要：目的探讨苏木精染色对冰冻切片组织RNA及蛋白质浓度和质量的影响。方法将10份乳腺癌组织冰冻切片进行苏木精染色以及苏木精染色后酸洗处理,利用显微切割技术从冰冻切片中获取特定乳腺癌细胞,Trizol提取相应的RNA及总蛋白,琼脂糖凝胶电泳和RT-PCR分析RNA质量,SDS-PAGE和Bandscan软件分析蛋白质电泳结果,并进行RNA和蛋白质的浓度测定。结果从10份乳腺癌组织冰冻切片中提取的RNA经琼脂糖凝胶电泳检测,苏木精染色组和苏木精染色后酸洗组的28S、18S、5S条带均有不同程度的降解,RT-PCR结果显示,RNA完整性较好;提取的蛋白电泳图谱处理前后无明显差异;苏木精染色以及苏木精染色后酸洗处理对乳腺癌组织中的RNA及蛋白质浓度无显著影响。结论苏木精染色对乳腺癌冰冻切片组织中RNA及蛋白质浓度和质量无显著影响,为肿瘤蛋白质组学的进一步研究提供了理论支持。Objective To explore the effect of hematoxylin staining on the concentrations and qualities of RNA and protein in frozen tissue section. Methods Ten frozen sections of breast cancer tissue were divided into 2 groups, 5 for each. The sections in one group were stained with hematoxylin, and those in the other group were stained with hematoxylin then washed with 2% hydrochloric acid, using the untreated sections as control. Specific breast cancer cells were collected from the sections by microdisseetion technique, from which total RNA and protein were extracted with Trizol reagent. Identify the extracted RNA by agarose gel electrophoresis and RT-PCR, and the protein by SDS-PAGE. Analyze the result of SDS-PAGE by Bandscan software and determine the concentrations of extracted RNA and protein. Results Agarose gel electrophoresis showed various degrees of degradation of 28S, 18S and 5S bands of RNA from the sections treated by hematoxylin staining as well as by hydrochloric acid washing after hematoxylin staining. RT-PCR showed good integrity of the RNA. The SDS-PAGE profiles of protein from the sections before and after treatment showed no significant difference. Neither hematoxylin staining nor hydrochloric acid washing after hematoxylin staining showed significant effect on RNA or protein concentration in breast cancer tissue. Conclusion Hematoxylin staining showed no significant effect on the concentrations and qualities of RNA and protein in frozen section of breast cancer tissue, which provided support in theory for further study on proteome of tumors.

关 键 词：苏木精染色 冰冻切片 显微切割 RNA 蛋白质 

分 类 号：Q2-33[生物学—细胞生物学]