Kapodistrias Nikolaos, Mavridis Konstantinos, Batistatou Anna, Gogou Penelope, Karavasilis Vasilios, Sainis Ioannis, Briasoulis Evangelos, Scorilas Andreas
Cancer Biobank Center, University of Ioannina, University Campus, Ioannina, Greece.
Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Panepistimiopolis, Athens, Greece.
Oncotarget. 2017 Jan 24;8(4):6896-6913. doi: 10.18632/oncotarget.14320.
Liposarcoma (LPS) is a malignancy with extreme heterogeneity and thus optimization towards personalizing patient prognosis and treatment is essential. Here, we evaluated miR-155, miR-21, miR-143, miR-145 and miR-451 that are implicated in LPS, as novel FFPE tissue biomarkers.A total of 83 FFPE tissue specimens from primary LPS and lipomas (LPM) were analyzed. A proteinase K incubation-Trizol treatment coupled protocol was used for RNA isolation. After polyadenylation of total RNA and reverse transcription, expression analysis of 9 candidate reference and 5 target miRNAs was performed by qPCR. Genorm and NormFinder were used for finding the most suitable molecules for normalization. Survival analyses were performed in order to evaluate the prognostic potential of miRNAs.MiR-103 and miR-191 are most suitable for normalization of miRNA expression in LPS. MiR-155 and miR-21 are clearly overexpressed (P<0.001) in LPS compared with LPM specimens, whereas miR-145 (P<0.001), miR-143 (P =0.008) and miR-451 (P=0.037) are underexpressed. MiR-155 (P=0.007) and miR-21 (P=0.029) are differentially expressed between well-differentiated, dedifferentiated, myxoid/round cell and pleomorphic LPs tumor subtypes. MiR-155 represents a novel independent indicator of unfavorable prognosis in LPS (HR = 2.97, 95% CI = 1.23-7.17, P = 0.016).
脂肪肉瘤（LPS）是一种具有高度异质性的恶性肿瘤，因此针对患者预后和治疗进行个性化优化至关重要。在此，我们评估了与脂肪肉瘤相关的miR-155、miR-21、miR-143、miR-145和miR-451，将其作为新型福尔马林固定石蜡包埋（FFPE）组织生物标志物。共分析了83份来自原发性脂肪肉瘤和脂肪瘤（LPM）的FFPE组织标本。采用蛋白酶K孵育-氯仿处理联合方案进行RNA提取。在对总RNA进行多聚腺苷酸化和逆转录后，通过定量聚合酶链反应（qPCR）对9种候选内参miRNA和5种靶miRNA进行表达分析。使用Genorm和NormFinder软件寻找最适合标准化的分子。进行生存分析以评估miRNA的预后潜力。MiR-103和miR-191最适合用于脂肪肉瘤中miRNA表达的标准化。与脂肪瘤标本相比，miR-155和miR-21在脂肪肉瘤中明显过表达（P<0.001），而miR-145（P<0.001）、miR-143（P =0.008）和miR-451（P=0.037）表达下调。MiR-155（P=0.007）和miR-21（P=0.029）在高分化、去分化、黏液样/圆形细胞和多形性脂肪肉瘤肿瘤亚型之间差异表达。MiR-155是脂肪肉瘤预后不良的一个新的独立指标（风险比=2.97，95%置信区间=1.23-7.17，P = 0.016）。
