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通过双标记逆转录聚合酶链反应检测黑色素瘤患者淋巴结清扫术后淋巴引流中的黑色素瘤细胞。

Detection of melanoma cells in the lymphatic drainage after lymph node dissection in melanoma patients by using two-marker reverse transcriptase-polymerase chain reaction assay.

作者信息

Włodzimierz Ruka, Rutkowski Piotr, Nowecki Zbigniew I, Kulik Jadwiga, Nasierowska-Guttmejer Anna, Siedlecki Janusz A

机构信息

Department of Soft Tissue/Bone Sarcoma and Melanoma, M. Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, W. Roentgena Str. 5, 02-781 Warsaw, Poland.

出版信息

Ann Surg Oncol. 2004 Nov;11(11):988-97. doi: 10.1245/ASO.2004.03.023. Epub 2004 Oct 15.

Abstract

BACKGROUND

The aim of this study was to evaluate the role of melanoma gene expression as a marker of the presence of melanoma cells in lymphatic drainage routinely collected after lymphadenectomy and to correlate reverse transcriptase-polymerase chain reaction (RT-PCR) assay results with recurrence, survival, and prognostic factors.

METHODS

We collected 24-hour postoperative lymphatic drainage samples (between days 2 and 4) from 93 patients with stage III melanoma who underwent radical lymphadenectomy between May 2002 and November 2003. We used RT-PCR assays with primers specific for the tyrosinase and MART-1 (Melan-A) genes. The samples were considered positive if at least one marker was expressed. Median follow-up time was 12.8 months.

RESULTS

In 18 (19.4%) of 93 patients, the RT-PCR assay results were positive: in 8 of 18 for tyrosinase only, in 7 of 18 for MART-1 only, and in 3 of 18 for both markers. We observed a significantly higher recurrence rate in patients with positive RT-PCR results (15 of 18; 83%) than negative results (26 of 75; 35%; P = .0001). Positive results of RT-PCR correlated with the number of involved lymph nodes (P = .0001) and extracapsular extension of nodal metastases (P = .03). We observed significant differences in overall and disease-free survival for RT-PCR-positive and -negative patients in univariate and multivariate analyses.

CONCLUSIONS

We observed positive RT-PCR assay results for melanoma cells in the lymphatic drainages of approximately 20% of patients after lymphadenectomy. This correlated significantly with early recurrence and shorter survival. These results may suggest that the RT-PCR assay could be useful for routinely analyzing postoperatively collected lymphatic drainage in stage III melanoma patients and for predicting disease progression.

摘要

背景

本研究的目的是评估黑色素瘤基因表达作为淋巴结清扫术后常规收集的淋巴引流液中黑色素瘤细胞存在标志物的作用,并将逆转录聚合酶链反应(RT-PCR)检测结果与复发、生存及预后因素相关联。

方法

我们收集了2002年5月至2003年11月期间接受根治性淋巴结清扫术的93例III期黑色素瘤患者术后24小时(术后第2至4天)的淋巴引流液样本。我们使用针对酪氨酸酶和MART-1(黑色素A)基因的特异性引物进行RT-PCR检测。如果至少一种标志物表达,则样本被视为阳性。中位随访时间为12.8个月。

结果

93例患者中有18例(19.4%)RT-PCR检测结果为阳性:18例中有8例仅酪氨酸酶阳性,18例中有7例仅MART-1阳性,18例中有3例两种标志物均阳性。我们观察到RT-PCR检测结果为阳性的患者复发率(18例中的15例;83%)显著高于阴性结果的患者(75例中的26例;35%;P = 0.0001)。RT-PCR阳性结果与受累淋巴结数量(P = 0.0001)和淋巴结转移的包膜外扩展(P = 0.03)相关。在单因素和多因素分析中,我们观察到RT-PCR阳性和阴性患者的总生存和无病生存存在显著差异。

结论

我们观察到淋巴结清扫术后约20%的患者淋巴引流液中黑色素瘤细胞的RT-PCR检测结果为阳性。这与早期复发和较短生存期显著相关。这些结果可能表明,RT-PCR检测可用于常规分析III期黑色素瘤患者术后收集的淋巴引流液,并预测疾病进展。

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