血清外泌体miR-125b,miR-21及miR-122对肝细胞癌诊断价值
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新疆维吾尔自治区卫生健康青年医学科技人才专项科研项目(WYWY-202010)


The diagnostic value of serum exosomal miR-125b,miR-21,miR-122 for hepatocellular carcinoma
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    摘要:

    摘要:目的 评估血清外泌体miR-125b,miR-21,miR-122,miR-375诊断肝细胞癌的效能,预测miR-125b潜在调控靶基因。方法 以健康体检人员50例和肝癌患者46例为研究对象,依次分为Health组和HCC组。收集临床资料,包括年龄、性别、甲胎蛋白(AFP)、α-L-岩藻糖苷酶(AFU)等指标。采集血清标本,分离纯化外泌体并鉴定,应用RT-qPCR技术检测各组血清外泌体miR-125b,miR-21,miR-122,miR-375的表达水平。进而行ROC 曲线分析,评估以上指标单独及联合检测对肝癌的诊断效能。基于miRNet 2.0在线网站预测miR-125b潜在靶基因,通过“ clusterProfiler” 分析间接推测其可能参与的信号转导通路。结果 提取纯化的血清样本中含有大量外泌体颗粒,与Health组相比,HCC组miR-125b,miR-21,miR-122表达水平上调(P<0.05);miR-375表达无显著差异(P>0.05)。 ROC曲线分析miR-125b,miR-21,miR-122,miR-375,AFP,AFU对HCC的诊断效能AUC值分别为0.900,0.857,0.827,0.597,0.881,0.733。联合检测miR-125b+AFP,miR-125b+miR-21, miR-125b+miR-21+AFP可将诊断效能分别提高至0.927,0.919,0.941。预测到64个与miR-125b具有潜在调控的靶基因,该miR-125b直接或间接参与p53信号通路,癌症中的miRNA,细胞衰老,TGF-beta信号通路等。结论miR-125b,miR-21,miR-122在肝癌患者血清外泌体中表达水平高于健康者,与AFP联合检测可提高诊断效能,有望成为肝癌诊断的候选标志物。

    Abstract:

    Abstract: Objective To evaluate the ffcacy of serum exosomal miR-125b, miR-21, miR- 122 and miR-375 in the diagnosis of hepatocellular carcinoma ( HCC),and predict the potential target genes regulated by miR-125b. Methods The clinical data of 46 HCC patients, 11 patients with chronic hepatitis B virus ( CHB) and 50 healthy controls, including age, gender, serum alpha fetoprotein ( AFP), 0- L-fucosidase ( AFU) and other indicators,were collected. The exosomes in their serum samples were isolated, purified and identified, and the expression levels of exosomal miR-125b, miR-21, miR-122 and miR -375 were detected by real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR ). The efficacies of individual and combined indicators for the diagnosis of HCC were evaluated by the ROC curve. The potential target genes of miR-125b were predicted by the miRTarBase. V8, TarBase. V8 and miRecords databases. The signal transduction pathways involved in miR- 125b were inferred by the analysis of clusterProfiler software. Results The extracted and purified serum samples contained a large number of exosome particles. The results of RT-qPCR showed that the expression levels of miR-125b, miR-21 and miR-122 in HCC patients were significantly higher than those in CHB patients and healthy controls ( P<0.05). The results of ROC curve analysis showed that the AUCHC values of miR-125b, miR-21, miR-122, miR-375 , AFP and AFU for the screening of HCC were 0.900, 0.857, 0.827, 0.597, 0.881 and 0.733, respectively. However, the AUCROC values of combined detection of miR -125b+AFP,miR-125b+miR-21 and miR -125b+ miR-21 + AFP for the diagnosis of HCC could increase to 0.927, 0.919 and 0.941, respectively. Nine target genes potentially regulated by miR- 125b,including TP53INPI, BAKI ,CEBPG, E2F3, ERBB2, ERBB3, NTRK3, SMO and SGPLI,were predicted by the database. The analysis of clusterProfiler software suggested that miR- 125b might directly or indirectly be involved in the p53 signalling pathway, miRNAs in cancer, cell aging, transforming growth factor (TGF)-β sigalling pathway, and etc. Conclusion The expression levels of miR-125b, miR-21 and miR-122 in serum exosomes of patients with HCC are increased, and their combination with AFP can improve the diagnostic efficiency of HCC, which may become the candidate markers for the diagnosis of HCC.

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王路,王魁,银花,叶莎,王娜.血清外泌体miR-125b, miR-21及miR-122对肝细胞癌诊断价值[J].临床检验杂志,2022,40(03):183-189

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  • 收稿日期:2021-09-15
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  • 在线发布日期: 2022-06-08
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