Evaluation of the predictive value of Her-2/neu gene expression on osteosarcoma therapy in laser-microdissected paraffin-embedded tissue
Evaluation of the predictive value of Her-2/neu gene expression on osteosarcoma therapy in laser-microdissected paraffin-embedded tissue
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DOI：
10.1038/labinvest.3700006
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发表时间：
2004-01-01
影响因子：
5
通讯作者：
Parsch, D
中科院分区：
文献类型：
--
作者：
Fellenberg, J;Krauthoff, A;Parsch, D
对化疗的组织学反应是目前高级别骨肉瘤最强的预后因素，但只能在治疗几周后才能评估。因此，诊断时检测化疗敏感性具有重要的临床意义。原癌基因 Her-2/neu 的表达已被证明对乳腺癌具有预测价值，也被认为是骨肉瘤的预后标志物，但主要是免疫组织化学研究的报道存在争议。因此，本研究的目的是研究激光显微切割骨肉瘤细胞中的 Her-2/neu 基因表达。激光显微切割能够从档案组织样本中精确分离形态明确的细胞，并与高度灵敏的实时 RT-PCR 技术相结合，成为细胞特异性基因表达分析的宝贵工具。通过对现有方案的优化，我们可以证明该技术可以成功应用于福尔马林固定、石蜡包埋和脱钙的骨肉瘤组织，具有高灵敏度和重现性。在分析的所有 17 个骨肉瘤活检组织中，我们可以检测到 Her-2/neu 基因表达。表达与术前化疗的反应显着相关，根据 Salzer-Kuntschik 六级量表进行组织学评估。基于 Her-2/neu 基因表达增加的风险评估与 17 例中的 16 例 (94%) 的组织学结果相符。这些数据证明了激光显微切割在基因表达分析中的可靠性，并表明 Her-2/neu 作为骨肉瘤治疗结果的预后标志物的可能作用。
Histologic response to chemotherapy is currently the strongest prognostic factor in high-grade osteosarcoma, but it can only be assessed after several weeks of therapy. Thus, detection of chemosensitivity at the time of diagnosis would be of great clinical importance. The expression of the proto-oncogene Her-2/neu has been shown to be of predictive value in breast cancer and has also been considered as prognostic marker for osteosarcomas, but reports of mainly immunohistochemical studies are controversial. Therefore, the aim of this study was to investigate Her-2/neu gene expression in laser-microdissected osteosarcoma cells. Laser microdissection enables the precise isolation of morphological defined cells from archival tissue specimens and is in combination with the highly sensitive real-time RT-PCR technique a valuable tool for cell-specific analysis of gene expression. Through optimization of current protocols, we could show that this technique can be successfully applied on formalin-fixed, paraffin-embedded and decalcified osteosarcoma tissue with high sensitivity and reproducibility. In all 17 osteosarcoma biopsies analyzed, we could detect Her-2/neu gene expression. Expression correlated significantly with the response to preoperative chemotherapy, which was assessed histologically according to the six-grade scale of Salzer-Kuntschik. Risk assessment on the basis of increased Her-2/neu gene expression matched the histologic findings in 16 out of 17 cases (94%). These data demonstrate the reliability of laser microdissection in the analysis of gene expression and suggest a possible role of Her-2/neu as prognostic marker for therapy outcome in osteosarcomas.