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使用ALU序列对转移性前列腺癌患者血浆游离DNA及其DNA完整性进行定量分析。

Quantitative analysis of plasma cell-free DNA and its DNA integrity in patients with metastatic prostate cancer using ALU sequence.

作者信息

Fawzy Amal, Sweify Karima M, El-Fayoumy Hany M, Nofal Nagwa

机构信息

Department of clinical and Chemical Pathology, National Cancer Institute, Cairo University, Egypt.

Department of Women's College for Arts, Science and Education, Ain Shams University, Egypt.

出版信息

J Egypt Natl Canc Inst. 2016 Dec;28(4):235-242. doi: 10.1016/j.jnci.2016.08.003. Epub 2016 Sep 12.

Abstract

BACKGROUND

Prostate cancer (PC) is the most common cancer affecting men, it accounts for 29% of all male cancer and 11% of all male cancer related death. DNA is normally released from an apoptotic source which generates small fragments of cell-free DNA, whereas cancer patients have cell-free circulating DNA that originated from necrosis, autophagy, or mitotic catastrophe, which produce large fragments.

AIM OF WORK

Differentiate the cell free DNA levels (cfDNA) and its integrity in prostate cancer patients and control group composed of benign prostate hyperplasia (BPH) and healthy persons.

METHODOLOGY

cf-DNA levels were quantified by real-time PCR amplification in prostate cancer patients (n= 50), (BPH) benign prostate hyperplasia (n= 25) and healthy controls (n= 30) using two sets of ALU gene (product size of 115bp and 247-bp) and its integrity was calculated as a ratio of qPCR results of 247bp ALU over 115bp ALU.

RESULTS

Highly significant levels of cf-DNA and its integrity in PC patients compared to BPH. Twenty-eight (56%) patients with prostate cancer had bone metastasis. ALU115 qpcr is superior to the other markers in discriminating metastatic patients with a sensitivity of 96.4% and a specificity of 86.4% and (AUC=0.981) CONCLUSION: ALU115 qpcr could be used as a valuable biomarker helping in identifying high risk patients, indicating early spread of tumor cells as a potential seed for future metastases.

摘要

背景

前列腺癌(PC)是影响男性的最常见癌症,占所有男性癌症的29%以及所有男性癌症相关死亡的11%。DNA通常从凋亡源释放,产生游离DNA小片段,而癌症患者有源自坏死、自噬或有丝分裂灾难的游离循环DNA,这些会产生大片段。

工作目的

区分前列腺癌患者与由良性前列腺增生(BPH)患者和健康人组成的对照组中的游离DNA水平(cfDNA)及其完整性。

方法

使用两组ALU基因(产物大小分别为115bp和247bp),通过实时PCR扩增对前列腺癌患者(n = 50)、良性前列腺增生(BPH)患者(n = 25)和健康对照者(n = 30)的cf-DNA水平进行定量,并将其完整性计算为247bp ALU与115bp ALU的qPCR结果之比。

结果

与BPH患者相比,PC患者的cf-DNA及其完整性水平非常显著。28名(56%)前列腺癌患者发生了骨转移。在鉴别转移患者方面,ALU115 qpcr优于其他标志物,敏感性为96.4%,特异性为86.4%,曲线下面积(AUC)=0.981。结论:ALU115 qpcr可作为一种有价值的生物标志物,有助于识别高危患者,提示肿瘤细胞早期扩散,这可能是未来转移的潜在根源。

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