Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study.
Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study.
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DOI：
10.1186/2043-9113-1-23
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发表时间：
2011-09-03
期刊：
影响因子：
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通讯作者：
Nishimura T
中科院分区：
文献类型：
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作者：
Nomura M;Fukuda T;Fujii K;Kawamura T;Tojo H;Kihara M;Bando Y;Gazdar AF;Tsuboi M;Oshiro H;Nagao T;Ohira T;Ikeda N;Gotoh N;Kato H;Marko-Varga G;Nishimura T
肺大细胞神经内分泌癌（LCNEC）是大细胞癌（LCC）的一个亚型，其特征在于具有小细胞肺癌（SCLC）共有的神经内分泌分化。LCNEC和SCLC之间的治疗前组织学区别迄今为止是有问题的，导致不良的临床结果。我们开始了一个项目，使用福尔马林固定石蜡包埋（FFPE）组织的蛋白质组学方法建立LCNEC的特征蛋白质靶点，这将有助于使诊断令人信服。通过激光显微切割从具有明确组织学诊断的LCNEC（n = 4）、SCLC（n = 5）和LCC（n = 5）的FFPE组织中的癌灶收集癌细胞。从收获的切片中提取蛋白质，胰蛋白酶消化，并进行HPLC/质谱分析。通过光谱计数对通过数据库检索鉴定的蛋白质进行半定量，并通过成对G统计进行统计分类。使用每组总共10个病例对结果进行化学验证，以确认蛋白质组学结果。对从三个癌症组鉴定的总共1981种蛋白质进行p < 0.05的成对G检验，并使用3D图检查蛋白质表达对LCNEC的特异性，其中坐标包含每两组比较的G统计值。与SCLC相比，我们鉴定了在LCNEC中优先表达的四种候选蛋白，具有令人信服的低p值：乙醛脱氢酶1家族成员A1（AL 1A 1）（p = 6.1 × 10-4）、醛酮还原酶家族1成员C1（AK 1C 1）（p = 9.6 × 10 -10）和C3（AK 1C 3）（p = 3.9 × 10 -10）和CD 44抗原（p = 0.021）。通过非参数精确推断检验证实了这些p值。有趣的是，所有这些候选者都属于癌症干细胞标记物。免疫组化支持蛋白质组学结果。这些结果表明，LCNEC的候选生物标志物与癌症干细胞相关，并且通过FFPE样品的这种蛋白质组学方法可有效地检测它们。
Large cell neuroendocrine carcinoma (LCNEC) of the lung, a subtype of large cell carcinoma (LCC), is characterized by neuroendocrine differentiation that small cell lung carcinoma (SCLC) shares. Pre-therapeutic histological distinction between LCNEC and SCLC has so far been problematic, leading to adverse clinical outcome. We started a project establishing protein targets characteristic of LCNEC with a proteomic method using formalin fixed paraffin-embedded (FFPE) tissues, which will help make diagnosis convincing. Cancer cells were collected by laser microdissection from cancer foci in FFPE tissues of LCNEC (n = 4), SCLC (n = 5), and LCC (n = 5) with definite histological diagnosis. Proteins were extracted from the harvested sections, trypsin-digested, and subjected to HPLC/mass spectrometry. Proteins identified by database search were semi-quantified by spectral counting and statistically sorted by pair-wise G-statistics. The results were immunohistochemically verified using a total of 10 cases for each group to confirm proteomic results. A total of 1981 proteins identified from the three cancer groups were subjected to pair-wise G-test under p < 0.05 and specificity of a protein's expression to LCNEC was checked using a 3D plot with the coordinates comprising G-statistic values for every two group comparisons. We identified four protein candidates preferentially expressed in LCNEC compared with SCLC with convincingly low p-values: aldehyde dehydrogenase 1 family member A1 (AL1A1) (p = 6.1 × 10-4), aldo-keto reductase family 1 members C1 (AK1C1) (p = 9.6x10-10) and C3 (AK1C3) (p = 3.9x10-10) and CD44 antigen (p = 0.021). These p-values were confirmed by non-parametric exact inference tests. Interestingly, all these candidates would belong to cancer stem cell markers. Immunohistochmistry supported proteomic results. These results suggest that candidate biomarkers of LCNEC were related to cancer stem cells and this proteomic approach via FFPE samples was effective to detect them.