创新 immunohistochemistry 识别在鼠科的 HCC 的侵略前面表示巨噬细胞的 MMP-9

• 作者: Christian Dierkes Elke Roeb Frank Lammert Jürgen Graf Martin Roderfeld Timo Rath
• 来源期刊: 世界肝病学杂志：英文版（电子版）
• 学科: 医学
• 关键词: Fluorescence STAINING HEPATOCELLULAR carcinoma Matrix METALLOPROTEINASE Tumor associated MACROPHAGES
• 年，卷（期）: 2010,（5）
• 页码范围: 175-179
• 页数: 5页
• 分类号: R735.7

摘要

AIM:To investigate the proteolytic contribution of tumor-associated macrophages(TAM)in tumor invasion,we analyzed whether TAM at the invasive front of small HCC in Abcb4-/--mice show an enhanced expression of MMP-9. METHODS:Liver cryosections of the hepatocellular carcinoma(HCC)invasive front from 12 mo old Abcb4-/--mice were stained for collagen typeⅠand MMP-9 using Alexa488 and Alexa568 labeled secondary antibodies.Afterwards,the Alexa568 dye was bleached and the macrophage marker F4/80 was visualized using Alexa568 labeled secondary antibodies.Finally, photographs of the invasive tumor front were digitally overlaid and analyzed. RESULTS:After complete bleaching of the primary dye,specific fluorescence staining of a third antigen, here F4/80,was successfully performed on the same histological section.With this method,we were able to identify conglomerates of matrix metalloproteinase (MMP-9)expressing macrophages within the tumor capsule of HCC. CONCLUSION:MMP-9 expressing macrophages are involved in matrix remodelling at the invasive tumor front of HCC.The described staining protocol provides a simple yet powerful extension of conventional immuno-histochemistry,facilitating visualization of at least three different antigens plus nuclei in one single histological section.