Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece.
Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece.
Genomics. 2019 Jul;111(4):642-652. doi: 10.1016/j.ygeno.2018.03.022. Epub 2018 Mar 31.
Tissue kallikrein, kallikrein-related peptidases (KLKs), and plasma kallikrein form the largest group of serine proteases in the human genome, sharing many structural and functional properties. Several KLK transcripts have been found aberrantly expressed in numerous human malignancies, confirming their prognostic or/and diagnostic values. However, the process of alternative splicing can now be studied in-depth due to the development of Next-Generation Sequencing (NGS). In the present study, we used NGS to discover novel transcripts of the KLK1 and KLK2 genes, after nested touchdown PCR. Bioinformatics analysis and PCR experiments revealed a total of eleven novel KLK transcripts (two KLK1 and nine KLK2 transcripts). In addition, the expression profiles of each novel transcript were investigated with nested PCR experiments using variant-specific primers. Since KLKs are implicated in human malignancies, qualifying as potential biomarkers, the quantification of the presented novel transcripts in human samples may have clinical applications in different types of cancer.
组织激肽释放酶、激肽释放酶相关肽酶（KLKs）和血浆激肽释放酶构成了人类基因组中最大的丝氨酸蛋白酶家族，它们具有许多结构和功能特性。许多 KLK 转录本在多种人类恶性肿瘤中异常表达，证实了它们的预后和/或诊断价值。然而，由于下一代测序（NGS）的发展，现在可以深入研究选择性剪接的过程。在本研究中，我们使用嵌套降落式 PCR 结合 NGS 技术发现 KLK1 和 KLK2 基因的新转录本。生物信息学分析和 PCR 实验共揭示了 11 种新的 KLK 转录本（2 种 KLK1 和 9 种 KLK2 转录本）。此外，使用变体特异性引物进行嵌套 PCR 实验研究了每个新转录本的表达谱。由于 KLKs 与人类恶性肿瘤有关，是潜在的生物标志物，因此在人类样本中定量检测这些新转录本可能在不同类型的癌症中有临床应用。
