Identification of canine circulating miRNAs as tumor biospecific markers using Next-Generation Sequencing and Q-RT-PCR.
Identification of canine circulating miRNAs as tumor biospecific markers using Next-Generation Sequencing and Q-RT-PCR.
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DOI：
10.1016/j.bbrep.2021.101106
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发表时间：
2021-12
影响因子：
2.7
通讯作者：
Smith BF
中科院分区：
文献类型：
--
作者：
Agarwal P;Crepps MP;Stahr NA;Kretzschmar WP;Harris HC;Prasad N;Levy SE;Smith BF
癌症诊断的延迟通常导致转移和无法成功治疗肿瘤。在早期阶段使用广泛的癌症特异性生物标志物可以改善癌症治疗和分期。这项研究探索了循环外泌体miRNA作为识别癌症患者的潜在诊断生物标志物。从13种犬癌细胞系（淋巴瘤、肥大细胞瘤、组织细胞细胞系、骨肉瘤、黑色素瘤和乳腺肿瘤）中分离分泌性外泌体miRNA，并通过下一代测序（NGS）进行测序。我们通过NGS鉴定了6种在所有癌细胞类型中升高的miRNAs（cfa-miR-9，-1841，-1306，-345，-132和-26b）。然后通过Q-RT-PCR检测通过NGS鉴定的miRNAs。PCR数据显示与NGS观察到的表达模式相似，但提供的倍数差异远低于NGS观察到的倍数差异。发现Cfa-miR-9是NGS和PCR中最一致升高的miRNA，使其成为最有可能证明诊断的miRNA。在这项研究中，我们已经证明，有可能在多种肿瘤类型中鉴定分泌升高的外泌体miRNA，这些外泌体miRNA可用作未来液体活检的循环诊断生物标志物。循环外泌体miRNA可用作早期癌症检测的诊断生物标志物。分泌型miRNA可以通过下一代测序进行测序和鉴定。在犬癌细胞系中有六种miRNAs升高。Q-RT-PCR用于分析和验证miRNAs的表达。在所有癌细胞类型中，Cfa-miR-9是最一致升高的miRNA。
Delay in cancer diagnosis often results in metastasis and an inability to successfully treat the tumor. The use of broadly cancer-specific biomarkers at an early stage may improve cancer treatment and staging. This study has explored circulatory exosomal miRNAs as potential diagnostic biomarkers to identify cancer patients. Secretory exosomal miRNAs were isolated from 13 canine cancer cell lines (lymphoma, mast cell tumor, histiocytic cell line, osteosarcoma, melanoma, and breast tumor) and were sequenced by Next-Generation sequencing (NGS). We have identified 6 miRNAs (cfa-miR-9, -1841, −1306, −345, −132, and −26b) by NGS that were elevated in all cancer cell types. The miRNAs identified by NGS were then examined by Q-RT-PCR. The PCR data demonstrated similar expression patterns to those seen with NGS but provided fold differences that were much lower than those seen for NGS. Cfa-miR-9 was found to be the most consistently elevated miRNA in NGS and PCR, making it the most likely miRNA to prove diagnostic. In this study, we have demonstrated that it is possible to identify exosomal miRNAs with elevated secretion across multiple tumor types that could be used as circulatory diagnostic biomarkers for liquid biopsy in the future. Circulatory exosomal miRNAs are useful as diagnostic biomarkers for early cancer detection. Secretory miRNAs can be sequenced and identified by Next-Generation sequencing. Six miRNAs were elevated in canine cancer cell lines. Q-RT-PCR was used to analyze and validate miRNAs expression. Cfa-miR-9 was most consistently elevated miRNAs in all cancer cell types.
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影响因子：
4.6
作者：
Heishima K;Ichikawa Y;Yoshida K;Iwasaki R;Sakai H;Nakagawa T;Tanaka Y;Hoshino Y;Okamura Y;Murakami M;Maruo K;Akao Y;Mori T
通讯作者：
Mori T
影响因子：
1.2
作者：
Noguchi, Shunsuke;Mori, Takashi;Maruo, Kohji
通讯作者：
Maruo, Kohji
影响因子：
46.9
作者：
Ma L;Reinhardt F;Pan E;Soutschek J;Bhat B;Marcusson EG;Teruya-Feldstein J;Bell GW;Weinberg RA
通讯作者：
Weinberg RA
影响因子：
2.8
作者：
Hu, Jinhua;Ruan, Jiugen;Xiong, Jianping
通讯作者：
Xiong, Jianping
影响因子：
3.7
作者：
Craig, Karlee K. L.;Wood, Geoffrey A.;Wood, R. Darren
通讯作者：
Wood, R. Darren