Laboratory of Investigative Dermatology, The Rockefeller University, New York, New York, USA.
Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD program, New York, New York, USA.
Pigment Cell Melanoma Res. 2024 Jan;37(1):81-89. doi: 10.1111/pcmr.13121. Epub 2023 Sep 30.
Melanoma accounts for the majority of skin cancer-related mortality, highlighting the need to better understand melanoma initiation and progression. In-depth molecular analysis of neoplastic melanocytes in whole tissue biopsies may be diluted by inflammatory infiltration, which may obscure gene signatures specific to neoplastic cells. Thus, a method is needed to precisely uncover molecular changes specific to tumor cells from a limited sample of primary melanomas. Here, we performed laser capture microdissection (LCM) and gene expression profiling of patient-derived frozen sections of pigmented lesions and primary cutaneous melanoma. Compared to bulk tissue analysis, analysis of LCM-derived samples identified 9528 additional differentially expressed genes (DEGs) including melanocyte-specific genes like PMEL and TYR, with enriched of pathways related to cell proliferation. LCM methodology also identified potentially targetable kinases specific to melanoma cells that were not detected by bulk tissue analysis. Taken together, our data demonstrate that there are marked differences in gene expression profiles depending on the method of sample isolation. We found that LCM captured higher expression of melanoma-related genes while whole tissue biopsy identified a wider range of inflammatory markers. Taken together, our data demonstrate that LCM is a valid approach to identify melanoma-specific changes using a relatively small amount of primary patient-derived melanoma sample.
黑色素瘤是皮肤癌相关死亡的主要原因,这凸显了更好地理解黑色素瘤发生和进展的必要性。对整个组织活检中的肿瘤性黑素细胞进行深入的分子分析可能会被炎症浸润所稀释,从而掩盖肿瘤细胞特有的基因特征。因此,需要一种方法来从有限的原发性黑素瘤样本中精确地揭示肿瘤细胞特有的分子变化。在这里,我们对患者来源的色素性病变和原发性皮肤黑素瘤的冷冻切片进行了激光捕获显微切割(LCM)和基因表达谱分析。与批量组织分析相比,LCM 衍生样本的分析鉴定出 9528 个额外的差异表达基因(DEGs),包括 PMEL 和 TYR 等黑素细胞特异性基因,富集了与细胞增殖相关的途径。LCM 方法还鉴定出了一些潜在的可针对肿瘤细胞的激酶,而这些激酶在批量组织分析中没有被检测到。总之,我们的数据表明,根据样品分离方法的不同,基因表达谱存在明显差异。我们发现,LCM 捕获了更高水平的与黑色素瘤相关的基因表达,而全组织活检则鉴定出了更广泛的炎症标志物。总之,我们的数据表明,LCM 是一种有效的方法,可以使用相对少量的原发性患者来源的黑素瘤样本来识别黑素瘤特异性变化。