鉴定血浆外泌体长链非编码RNA作为胃癌早期诊断的生物标志物
Identification of plasma exosomal lncRNA as a biomarker for early diagnosis of gastric cancer.
作者信息
Wei Ye, Hu Xuming, Yuan Shuai, Zhao Yue, Zhu Chunhui, Guo Mingzhou, Cui Hengmi
机构信息
College of Medicine, Yangzhou University, Yangzhou, China.
Institute of Epigenetics and Epigenomics and College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
出版信息
Front Genet. 2024 Oct 8;15:1425591. doi: 10.3389/fgene.2024.1425591. eCollection 2024.
BACKGROUND
There were about 1,090,000 gastric cancer (GC) cases in 2020 in China. The incidence and mortality rates ranked the fifth and third among all kinds of cancers in China. Early diagnosis plays an important role in the treatment and prognosis of gastric cancer. In recent years, noninvasive diagnosis, especially plasma exosome lncRNAs, has become a promissing biomarkers with high specificity and sensitivity for early diagnosis of cancers.
METHODS
In this study, plasma exosomes of patients with early gastric cancer were extracted efficiently by affinity membrane separation technology, including affinity adsorption, elution, affinity membrane regeneration and other steps. After identified by electron microscopy observation, particle size analysis and Western blot verification, the lncRNAs in the exosomes were extracted and were analysized by high-throughput RNA sequencing (RNA-Seq). The differentially expressed lncRNAs were verified by RT-qPCR in 93 patients with early gastric cancer and 49 normal controls.
RESULTS
Electron microscopy, particle size analysis and Western blot showed that exosomes were successfully isolated from plasma. RNA-Seq results show that 76 lncRNAs were upregulated and 260 lncRNAs were downregulated in plasma exosomes of early gastric cancer patients compared with normal controls. RT-qPCR analysis indicated that a total of 6 lncRNAs were significantly and differentially expressed in gastric cancer patients compared to normal controls, with 2 (lncmstrg. 1319590, Lncmstrg. 2312697) highly expressed and 4 lowly expressed (lncmstr-g.1004024.1, lncmstrg. 2441832.8, lncmstrg. 315376.1, lncmstrg.907985.2,) ( < 0.05). The survival curve analysis indicated that lncmstrg.2441832.8 and lncmstrg.2312697 had higher sensitivity and specificity for the diagnosis of gastric cancer, respectively and AUC curve areas were 0.6211 and 0.631, < 0.05, respectively, which were greater than the traditional clinical detection indexes CEA (0.61) and AFP (0.57). When combined lncmstrg.2441832.8 and lncmstrg.2312697 in gastric cancer diagnosis, AUC curve area reached 0.73, which was greater than CA199 (0.71).
CONCLUSION
Lncmstrg.2441832.8 and lncmstrg.2312697 may be a potential and promissing biomarkers for early diagnosis of gastric cancer.
背景
2020年中国胃癌病例约109万例。其发病率和死亡率在中国各类癌症中分别位居第五和第三。早期诊断对胃癌的治疗和预后起着重要作用。近年来，非侵入性诊断，尤其是血浆外泌体长链非编码RNA（lncRNAs），已成为具有高特异性和敏感性的、有望用于癌症早期诊断的生物标志物。
方法
在本研究中，采用亲和膜分离技术高效提取早期胃癌患者的血浆外泌体，包括亲和吸附、洗脱、亲和膜再生等步骤。经电子显微镜观察、粒径分析和蛋白质免疫印迹验证后，提取外泌体中的lncRNAs，并通过高通量RNA测序（RNA-Seq）进行分析。在93例早期胃癌患者和49例正常对照中，采用逆转录定量聚合酶链反应（RT-qPCR）验证差异表达的lncRNAs。
结果
电子显微镜、粒径分析和蛋白质免疫印迹结果表明，成功从血浆中分离出外泌体。RNA-Seq结果显示，与正常对照相比，早期胃癌患者血浆外泌体中有76个lncRNAs上调，260个lncRNAs下调。RT-qPCR分析表明，与正常对照相比，共有6个lncRNAs在胃癌患者中显著差异表达，其中2个（lncmstrg.1319590、Lncmstrg.2312697）高表达，4个低表达（lncmstr-g.1004024.1、lncmstrg.2441832.8、lncmstrg.315376.1、lncmstrg.907985.2）（<0.05）。生存曲线分析表明，lncmstrg.2441832.8和lncmstrg.2312697对胃癌诊断分别具有较高的敏感性和特异性，曲线下面积（AUC）分别为0.6211和0.631（<0.05），均大于传统临床检测指标癌胚抗原（CEA，0.61）和甲胎蛋白（AFP，0.57）。当联合lncmstrg.2441832.8和lncmstrg.2312697用于胃癌诊断时，AUC曲线面积达到0.73，大于糖类抗原199（CA199，0.71）。
结论
lncmstrg.2441832.8和lncmstrg.2312697可能是早期诊断胃癌的潜在且有前景的生物标志物。
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