Department of Pathology, Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow, U.P., 226010, India.
Department of Biosciences, Integral University, Lucknow, U.P., 226026, India.
Pathol Oncol Res. 2019 Jul;25(3):925-936. doi: 10.1007/s12253-017-0380-6. Epub 2018 Jan 28.
The current study investigates the role of circulating free DNA (cfDNA) as a liquid biopsy in diagnosis gall bladder carcinoma (GBC) utilizing levels of long DNA fragments (ALU247) derived from tumor necrosis, short apoptotic fragments (ALU115) denoting total cfDNA and cfDNA integrity denoting ratio of ALU247 and ALU115. The global methylation status of cfDNA was also estimated with the hypothesis that these parameters provide a diagnostic distinction between cancer and non-cancer subjects, with higher or altered values favoring presence of malignancy. Study group included 60 cases of GBC and 36 controls including diseased controls (cholecystitis) and healthy subjects. Median levels of ALU115, ALU247 and cfDNA integrity were significantly different in GBC at 1790.88, 673.75, 0.4718 vs. controls at 840.73, 165.03, 0.1989 ng/ml respectively. Global DNA methylation was not significantly different between GBC at 0.679% and controls at 0.695%. The sensitivity and specificity of ALU 247 in discriminating GBC from controls was highest with a sensitivity, specificity and diagnostic accuracy of 80.0%, 86.1% and 82.2% respectively. Global DNA methylation showed lowest sensitivity of 55.0% and specificity of 50.0%. Clinico-pathological parameters showing significant association with cfDNA integrity, on ROC curve analysis, showed significant diagnostic discrimination of the tumor stage, lymphovascular invasion, disease stage and grade histology. This is a first time analysis of ALU115, ALU247 and cfDNA integrity in the diagnosis of GBC and confirms that the combination of ALU247 and cfDNA integrity provides good sensitivity, specificity and diagnostic accuracy in discriminating GBC from controls as well correlates with aggressive disease parameters.
本研究利用源自肿瘤坏死的长 DNA 片段（ALU247）、表示总 cfDNA 的短凋亡片段（ALU115）和表示 cfDNA 完整性的 ALU247 与 ALU115 比值，研究循环游离 DNA（cfDNA）作为液体活检在诊断胆囊癌（GBC）中的作用。还估计了 cfDNA 的整体甲基化状态，假设这些参数提供了癌症与非癌症患者之间的诊断区别，较高或改变的值有利于恶性肿瘤的存在。研究组包括 60 例 GBC 和 36 例对照，包括患病对照（胆囊炎）和健康对照。在 GBC 中，ALU115、ALU247 和 cfDNA 完整性的中位数水平分别为 1790.88、673.75 和 0.4718，而对照组分别为 840.73、165.03 和 0.1989ng/ml。GBC 与对照组之间的整体 DNA 甲基化没有显著差异，分别为 0.679%和 0.695%。ALU247 区分 GBC 与对照组的敏感性和特异性最高，敏感性、特异性和诊断准确性分别为 80.0%、86.1%和 82.2%。整体 DNA 甲基化的敏感性最低为 55.0%，特异性为 50.0%。与 cfDNA 完整性具有显著关联的临床病理参数在 ROC 曲线分析中显示出对肿瘤分期、血管侵犯、疾病分期和组织学分级的显著诊断区分。这是首次分析 ALU115、ALU247 和 cfDNA 完整性在 GBC 诊断中的应用，证实 ALU247 和 cfDNA 完整性的组合在区分 GBC 与对照组方面具有良好的敏感性、特异性和诊断准确性，并且与侵袭性疾病参数相关。
