原发性皮肤黑色素瘤的基因表达谱分析与临床结果

Gene expression profiling of primary cutaneous melanoma and clinical outcome.

作者信息

Winnepenninckx Véronique, Lazar Vladimir, Michiels Stefan, Dessen Philippe, Stas Marguerite, Alonso Soledad R, Avril Marie-Françoise, Ortiz Romero Pablo L, Robert Thomas, Balacescu Ovidiu, Eggermont Alexander M M, Lenoir Gilbert, Sarasin Alain, Tursz Thomas, van den Oord Joost J, Spatz Alain

机构信息

Department of Pathology, Gustave-Roussy Institute, 94805 Villejuif Cedex, France.

出版信息

J Natl Cancer Inst. 2006 Apr 5;98(7):472-82. doi: 10.1093/jnci/djj103.

DOI:10.1093/jnci/djj103

PMID:16595783

Abstract

BACKGROUND

Gene expression profiling data for human primary cutaneous melanomas are scarce because of the lack of retrospective collections of frozen tumors. To identify differentially expressed genes that may be involved in melanoma progression and prognosis, we investigated the relationship between gene expression profiles and clinical outcome in a cohort of patients with primary melanoma.

METHODS

Labeled complementary RNA (cRNA) from each tissue sample was hybridized to a pangenomic 44K 60-mer oligonucleotide microarray. Class comparison and class prediction analyses were performed to identify genes whose expression in primary melanomas was associated with 4-year distant metastasis-free survival among 58 patients with at least 4 years of follow-up, distant metastasis, or death. Results were validated immunohistochemically at the protein level in 176 independent primary melanomas from patients with a median clinical follow-up of 8.5 years. Survival was analyzed with a Cox multivariable model and stratified log-rank test. All statistical tests were two-sided.

RESULTS

We identified 254 genes that were associated with distant metastasis-free survival of patients with primary melanoma. These 254 genes include genes involved in activating DNA replication origins, such as minichromosome maintenance genes and geminin. Twenty-three of these genes were studied at the protein level; expression of five (MCM4, P = .002; MCM3, P = .030; MCM6, P = .004; KPNA2, P = .021; and geminin, P = .004) was statistically significantly associated with overall survival in the validation set. In a multivariable Cox model adjusted for tumor thickness, ulceration, age, and sex, expression of MCM4 (hazard ratio [HR] of death = 4.04, 95% confidence interval [CI] = 1.39 to 11.76; P = .010) and MCM6 (HR of death = 7.42, 95% CI = 1.99 to 27.64; P = .003) proteins was still statistically significantly associated with overall survival.

CONCLUSION

We identified 254 genes whose expression was associated with metastatic dissemination of cutaneous melanomas. These genes may shed light on the molecular mechanisms underlying poor prognosis in melanoma patients.

摘要

背景

由于缺乏对冷冻肿瘤的回顾性收集，人类原发性皮肤黑色素瘤的基因表达谱数据稀缺。为了鉴定可能参与黑色素瘤进展和预后的差异表达基因，我们在一组原发性黑色素瘤患者中研究了基因表达谱与临床结局之间的关系。

方法

将来自每个组织样本的标记互补RNA（cRNA）与全基因组44K 60聚体寡核苷酸微阵列杂交。进行类别比较和类别预测分析，以鉴定在至少随访4年、发生远处转移或死亡的58例患者中，其在原发性黑色素瘤中的表达与4年无远处转移生存率相关的基因。在176例临床随访时间中位数为8.5年的患者的独立原发性黑色素瘤中，在蛋白质水平通过免疫组织化学对结果进行验证。采用Cox多变量模型和分层对数秩检验分析生存率。所有统计检验均为双侧检验。

结果

我们鉴定出254个与原发性黑色素瘤患者无远处转移生存率相关的基因。这254个基因包括参与激活DNA复制起点的基因，如微型染色体维持基因和geminin。其中23个基因在蛋白质水平进行了研究；其中5个基因（MCM4，P = 0.002；MCM3，P = 0.030；MCM6，P = 0.004；KPNA2，P = 0.021；geminin，P = 0.004）的表达与验证集中的总生存率在统计学上显著相关。在根据肿瘤厚度、溃疡、年龄和性别进行调整的多变量Cox模型中，MCM4（死亡风险比[HR]=4.04，95%置信区间[CI]=1.39至11.76；P = 0.010）和MCM6（死亡HR = 7.42，95%CI = 1.99至27.64；P = 0.003）蛋白的表达仍与总生存率在统计学上显著相关。