Gore S D, Weng L J, Burke P J
Johns Hopkins Oncology Center, Baltimore, MD 21287-8963.
Exp Hematol. 1993 Dec;21(13):1702-8.
Despite its growing use as a marker of proliferation, the relationship between expression of the nuclear antigen Ki67 and other indices of proliferation in acute myelogenous leukemia (AML) has not been elucidated. In this study, short-term primary suspension cultures of human AML bone marrow cells were used to compare flow-cytometric methods of quantifying Ki67 expression and to test whether flow-cytometric determination of Ki67 expression correlates with incorporation of 3H-thymidine or bromodeoxyuridine (BrdU). BrdU incorporation was determined by staining of cells with anti-BrdU and propidium iodide (PI) followed by flow cytometry. When samples were double-labeled with Ki67 and PI, Ki67 was underestimated compared to single-color quantification of the nuclear antigen. Ki67+ cell number correlated well with incorporation of 3H-thymidine (r = 0.89, p < 0.001). Cells from 17 cases of AML were cultured for 3 days in the presence and absence of a variety of growth factors and growth factor combinations before comparison of Ki67 expression and BrdU incorporation. Ki67 expression correlated strongly with BrdU incorporation (r = 0.82, p < 0.001). Unlike the double-label with PI, containing with a phycoerythrin (PE)-labeled antibody directed against a cell surface antigen did not significantly affect Ki67 quantification. Flow cytometric determination of Ki67 is a simple and valid measurement of proliferation in AML bone marrow cells.
尽管核抗原Ki67作为增殖标志物的应用日益广泛，但急性髓性白血病（AML）中Ki67表达与其他增殖指标之间的关系尚未阐明。在本研究中，使用人AML骨髓细胞的短期原代悬浮培养物来比较定量Ki67表达的流式细胞术方法，并测试Ki67表达的流式细胞术测定是否与3H-胸腺嘧啶核苷或溴脱氧尿苷（BrdU）的掺入相关。通过用抗BrdU和碘化丙啶（PI）对细胞进行染色，然后进行流式细胞术来测定BrdU掺入。当样品用Ki67和PI进行双重标记时，与核抗原的单色定量相比，Ki67被低估。Ki67+细胞数与3H-胸腺嘧啶核苷的掺入密切相关（r = 0.89，p < 0.001）。在比较Ki67表达和BrdU掺入之前，将17例AML患者的细胞在有和没有多种生长因子及生长因子组合的情况下培养3天。Ki67表达与BrdU掺入密切相关（r = 0.82，p < 0.001）。与用PI进行双重标记不同，用针对细胞表面抗原的藻红蛋白（PE）标记抗体进行双重标记不会显著影响Ki67定量。流式细胞术测定Ki67是一种简单有效的AML骨髓细胞增殖测量方法。
