Goethals Laurence, Debucquoy Annelies, Perneel Christiaan, Geboes Karel, Ectors Nadine, De Schutter Harlinde, Penninckx Freddy, McBride William H, Begg Adrian C, Haustermans Karin M
Department of Radiation Oncology, University Hospital Gasthuisberg, Leuven, Belgium.
Int J Radiat Oncol Biol Phys. 2006 May 1;65(1):246-54. doi: 10.1016/j.ijrobp.2006.01.007.
To detect and quantify hypoxia in colorectal adenocarcinomas by use of pimonidazole and iododeoxyuridine (IdUrd) as extrinsic markers and carbonic anhydrase IX (CA IX), microvessel density (MVD), epidermal growth-factor receptor (EGFR), and vascular endothelial growth factor (VEGF) as intrinsic markers of hypoxia.
Twenty patients with an adenocarcinoma of the left colon and rectum treated by primary surgery were injected with pimonidazole and IdUrd. Serial sections of tumor biopsies were single stained for VEGF, EGFR, Ki67, and double stained for blood vessels in combination with either pimonidazole, IdUrd, or CA IX. Percentage of expression was scored as well as colocalization of pimonidazole with CA IX.
The median percentage of hypoxia, as judged by pimonidazole staining, was 16.7% (range, 0-52.4%). The expression of pimonidazole correlated inversely with the total MVD and endothelial cord MVD (R = -0.55, p = 0.01; R = -0.47, p = 0.04). Good colocalization was found between pimonidazole and CA IX in only 30% of tumors, with no correlation overall between pimonidazole and CA IX, VEGF, or EGFR or between the different intrinsic markers. Cells around some vessels (0.08-11%) were negative for IdUrd but positive for Ki 67, which indicated their lack of perfusion at the time of injection.
Chronic and acute hypoxic regions are present in colorectal tumors, as shown by pimonidazole and IdUrd staining. Only in a minority of tumors did an association exist between the areas stained by pimonidazole and those positive for CA IX. Pimonidazole also did not correlate with expression of other putative intrinsic hypoxia markers (VEGF, EGFR).
使用匹莫硝唑和碘脱氧尿苷（IdUrd）作为外源性标志物，以及碳酸酐酶IX（CA IX）、微血管密度（MVD）、表皮生长因子受体（EGFR）和血管内皮生长因子（VEGF）作为缺氧的内源性标志物，检测并量化结肠直肠癌中的缺氧情况。
对20例接受一期手术治疗的左半结肠癌和直肠癌患者注射匹莫硝唑和IdUrd。肿瘤活检的连续切片分别单独进行VEGF、EGFR、Ki67染色，并将血管与匹莫硝唑、IdUrd或CA IX进行双重染色。对表达百分比进行评分，并对匹莫硝唑与CA IX的共定位情况进行评估。
通过匹莫硝唑染色判断，缺氧的中位百分比为16.7%（范围为0 - 52.4%）。匹莫硝唑的表达与总MVD和内皮条索MVD呈负相关（R = -0.55，p = 0.01；R = -0.47，p = 0.04）。仅在30%的肿瘤中发现匹莫硝唑与CA IX有良好的共定位，总体上匹莫硝唑与CA IX、VEGF或EGFR之间以及不同内源性标志物之间均无相关性。一些血管周围的细胞（0.08 - 11%）IdUrd染色为阴性，但Ki67染色为阳性，这表明它们在注射时缺乏灌注。
匹莫硝唑和IdUrd染色显示，结肠直肠癌中存在慢性和急性缺氧区域。只有少数肿瘤中，匹莫硝唑染色区域与CA IX阳性区域之间存在关联。匹莫硝唑也与其他假定的内源性缺氧标志物（VEGF、EGFR）的表达无关。
