Department of Ophthalmology, Oslo University Hospital HF, Oslo, Norway.
Acta Ophthalmol. 2013 Jun;91(4):343-8. doi: 10.1111/j.1755-3768.2012.02449.x. Epub 2012 May 25.
Approximately 50% of patients with uveal melanomas develop metastases. Thus, it is important to improve our understanding of how melanoma metastases develop.
As part of a uveal melanoma micrometastasis study, we compared the detection rates of immunomagnetically selected (IMS) tumour cells in bone marrow (BM) with positively stained tumour cells using immunocytochemistry (ICC). Bone marrow mononuclear cells were isolated. Immunocytochemistry cytospin preparations were immunocytochemically stained in parallel with two different melanoma antibodies, 9.2.27 and HMB45. Using IMS, melanoma cells were selected from BM mononuclear cell fractions using immunomagnetic beads coated with the 9.2.27 antibody and identified by light microscopy.
In cytospin preparations from 226 patients, melanoma cells were detected in 24 (10.6%), 10 with 9.2.27 and 17 with the HMB45 antibody. In three cases, we found positive cells with both antibodies. Six of the 226 (2.6%) patients that stained positively with ICC died with metastatic disease, all also positive with IMS. Sixty-six (29.2%) patients had positive BM samples with IMS at the first examination. Immunomagnetic selection (IMS) was positive in 36.8% of the 57 patients who later developed clinical metastases. Twenty-one IMS-positive patients and 31 IMS-negative patients died of metastases, in total 52 of 226 patients (23.0%). The mortality rate of melanoma metastasis was 24% (6/24) after at least 4 ½ years in ICC-positive patients compared to 38.5% (20/52) in IMS-positive patients.
The presence of melanoma cells in BM of patients with uveal melanoma is documented in our study with IMS and ICC. Immunomagnetically selected is more sensitive than ICC in detecting tumour cells in BM. However, statistically, we did not find any prognostic impact of the presence of melanoma cells in BM.
约 50%的葡萄膜黑色素瘤患者会发生转移。因此，提高我们对黑色素瘤转移发生机制的认识非常重要。
作为葡萄膜黑色素瘤微转移研究的一部分，我们比较了免疫磁珠选择（IMS）和免疫细胞化学（ICC）法在骨髓（BM）中检测肿瘤细胞的检出率。分离 BM 单个核细胞，平行进行 ICC 细胞免疫荧光染色。使用 IMS，用 9.2.27 抗体包被的免疫磁珠从 BM 单个核细胞中选择黑色素瘤细胞，并通过光学显微镜进行鉴定。
在 226 例患者的细胞涂片制备中，用 9.2.27 抗体检测到 10 例（10.6%）、用 HMB45 抗体检测到 17 例（17.0%）患者存在黑色素瘤细胞，在这 3 例患者中，我们发现这两种抗体均呈阳性的细胞。在 226 例 ICC 阳性患者中，有 6 例死亡并发生转移，且均为 IMS 阳性。首次检查时，66 例（29.2%）患者的 BM 样本 IMS 阳性。在随后发生临床转移的 57 例患者中，有 36.8%（21/57）的患者 IMS 阳性。21 例 IMS 阳性患者和 31 例 IMS 阴性患者死于转移，226 例患者中共有 52 例（23.0%）。在 ICC 阳性患者中，至少随访 4 年半后，黑色素瘤转移的死亡率为 24%（6/24），而 IMS 阳性患者为 38.5%（20/52）。
本研究通过 IMS 和 ICC 证实了黑色素瘤患者 BM 中黑色素瘤细胞的存在。与 ICC 相比，免疫磁珠选择法在检测 BM 中的肿瘤细胞方面更敏感。然而，统计学上，我们并未发现 BM 中黑色素瘤细胞的存在具有任何预后影响。
