Skeparnias Ilias, Anastasakis Dimitrios, Grafanaki Katerina, Kyriakopoulos George, Alexopoulos Panagiotis, Dougenis Dimitrios, Scorilas Andreas, Kontos Christos K, Stathopoulos Constantinos
Department of Biochemistry, School of Medicine, University of Patras, 26504 Patras, Greece.
National Institute of Musculoskeletal and Arthritis and Skin Disease, NIH, 50 South Drive, Room 1152, Bethesda, MD 20892, USA.
Cancers (Basel). 2020 Oct 20;12(10):3056. doi: 10.3390/cancers12103056.
Transcriptomics profiles of miRNAs, tRNAs or tRFs are used as biomarkers, after separate examination of several cancer cell lines, blood samples or biopsies. However, the possible contribution of all three profiles on oncogenic signaling and translation as a net regulatory effect, is under investigation. The present analysis of miRNAs and tRFs from lung cancer biopsies indicated putative targets, which belong to gene networks involved in cell proliferation, transcription and translation regulation. In addition, we observed differential expression of specific tRNAs along with several tRNA-related genes with possible involvement in carcinogenesis. Transfection of lung adenocarcinoma cells with two identified tRFs and subsequent NGS analysis indicated gene targets that mediate signaling and translation regulation. Broader analysis of all major signaling and translation factors in several biopsy specimens revealed a crosstalk between the PI3K/AKT and MAPK pathways and downstream activation of eIF4E and eEF2. Subsequent polysome profile analysis and 48S pre-initiation reconstitution experiments showed increased global translation rates and indicated that aberrant expression patterns of translation initiation factors could contribute to elevated protein synthesis. Overall, our results outline the modulatory effects that possibly correlate the expression of important regulatory non-coding RNAs with aberrant signaling and translation deregulation in lung cancer.
在对几种癌细胞系、血液样本或活检组织进行单独检测后,微小RNA(miRNA)、转运RNA(tRNA)或tRNA衍生片段(tRF)的转录组学图谱被用作生物标志物。然而,这三种图谱对致癌信号传导和翻译的总体调节作用仍在研究之中。目前对肺癌活检组织中miRNA和tRF的分析表明了推定的靶标,这些靶标属于参与细胞增殖、转录和翻译调控的基因网络。此外,我们观察到特定tRNA以及几个可能参与致癌过程的tRNA相关基因的差异表达。用两种已鉴定的tRF转染肺腺癌细胞并随后进行二代测序(NGS)分析,确定了介导信号传导和翻译调控的基因靶标。对多个活检样本中所有主要信号传导和翻译因子的更广泛分析揭示了PI3K/AKT和MAPK信号通路之间的相互作用以及真核翻译起始因子4E(eIF4E)和真核延伸因子2(eEF2)的下游激活。随后的多核糖体图谱分析和48S前起始复合物重建实验表明整体翻译速率增加,并表明翻译起始因子的异常表达模式可能导致蛋白质合成增加。总体而言,我们的结果概述了重要调控性非编码RNA的表达与肺癌中异常信号传导和翻译失调之间可能存在的调节作用。