Dawson A E, Norton J A, Weinberg D S
Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts.
Am J Pathol. 1990 May;136(5):1115-24.
Although tumor DNA content and proliferation are usually determined by flow cytometry (FCM), quantitative microscopic image analysis is a viable alternative technique that also provides important histologic correlations. To compare these methods, we measured DNA content and proliferation in 54 consecutive breast cancers and 15 benign breast lesions by FCM and IA. DNA content determination was concordant in 49 of 54 cancers measured by FCM and IA. Four of the discordant cases were aneuploid by IA and diploid by FCM. There was good correlation between the DNA index (DI) measured by FCM and IA (r = 0.89, P less than 0.0001). Proliferation was assessed by IA quantitation of Ki-67 and PCNA/Cyclin antibody staining, as well as by flow cytometric S-phase fraction (SPF). Ki-67 positivity was greater in breast cancer than in benign controls (21.6% +/- 13.1% vs. 7.9% +/- 5.6% [P less than 0.0001]), as was PCNA/Cyclin positivity (10.2 +/- 6.7% vs. 2.7 +/- 2.5% [P less than 0.0001]). S-phase fraction measured by FCM was 7.9% +/- 5.7% for carcinomas and 3.17% +/- 2.1% for benign controls (P less than 0.003). Ki-67 and Cyclin staining, as well as SPF, were significantly increased in aneuploid compared to diploid tumors, and increased staining was associated with worsening nuclear grade. There were significant correlations between SPF and Ki-67 staining (r = 0.48, P less than 0.0001) and SPF and Cyclin staining (r = 0.48, P less than 0.0001). We conclude that FCM and IA provide comparable measurements of DNA content, although occasional discrepancies occur. Image analysis provides a valuable alternative method for assessing tumor cell proliferation and may offer certain advantages over FCM.
虽然肿瘤DNA含量和增殖情况通常通过流式细胞术（FCM）来测定，但定量显微图像分析也是一种可行的替代技术，它还能提供重要的组织学相关性。为比较这些方法，我们通过FCM和图像分析（IA）测定了54例连续乳腺癌和15例乳腺良性病变的DNA含量及增殖情况。在通过FCM和IA测定的54例癌症中，有49例的DNA含量测定结果一致。4例不一致的病例经IA测定为非整倍体，经FCM测定为二倍体。FCM和IA所测的DNA指数（DI）之间存在良好的相关性（r = 0.89，P小于0.0001）。通过IA对Ki-67和PCNA/细胞周期蛋白抗体染色进行定量，以及通过流式细胞术测定S期细胞分数（SPF）来评估增殖情况。乳腺癌中Ki-67阳性率高于良性对照（21.6%±13.1%对7.9%±5.6%[P小于0.0001]），PCNA/细胞周期蛋白阳性率也是如此（10.2±6.7%对2.7±2.5%[P小于0.0001]）。FCM测定的癌组织S期细胞分数为7.9%±5.7%，良性对照为3.17%±2.1%（P小于0.003）。与二倍体肿瘤相比，非整倍体肿瘤的Ki-67和细胞周期蛋白染色以及SPF均显著增加，且染色增加与核分级恶化相关。SPF与Ki-67染色之间（r = 0.48，P小于0.0001）以及SPF与细胞周期蛋白染色之间（r = 0.48，P小于0.0001）存在显著相关性。我们得出结论，虽然偶尔会出现差异，但FCM和IA对DNA含量的测定结果具有可比性。图像分析为评估肿瘤细胞增殖提供了一种有价值的替代方法，可能比FCM具有某些优势。
