Unveiling novel targets of paclitaxel resistance by single molecule long-read RNA sequencing in breast cancer
Unveiling novel targets of paclitaxel resistance by single molecule long-read RNA sequencing in breast cancer
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通过单分子长读长 RNA 测序揭示乳腺癌紫杉醇耐药性的新靶点
DOI：
10.1038/s41598-019-42184-z
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发表时间：
2019-04-15
影响因子：
4.6
通讯作者：
Shao, Zhi-ming
中科院分区：
文献类型：
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作者：
Lian, Bi;Hu, Xin;Shao, Zhi-ming
RNA测序已成为研究癌症转录本的最常用技术之一，但其长度限制了其在选择性剪接(AS)事件和新的异构体上的应用。首先，我们对野生型(231-WT)和紫杉醇耐药型(231-PTX)的人乳腺癌细胞MDA-MBA-231进行了单分子长读RNA测序(ISO-SEQ)和重组短读RNA测序(RNA-SEQ)。这两种测序技术既提供了准确的转录本序列，又提供了深入的转录本覆盖。然后，我们结合短读和长读的RNA-seq来分析替代事件和新的异构体。最后，我们选择BAK1作为我们的候选目标来验证我们的分析。我们的结果表明，为了更好地表征癌症基因组功能，可能需要应用单分子长读RNA测序来获得更深层次和更准确的转录水平信息。我们的结果表明，为了更好地表征癌症基因组功能，可能需要应用单分子长读RNA测序来获得更深层次和更准确的转录水平视图。
RNA sequencing has become one of the most common technology to study transcriptomes in cancer, whereas its length limits its application on alternative splicing (AS) events and novel isoforms. Firstly, we applied single molecule long-read RNA sequencing (Iso-seq) and de novo assembly with short-read RNA sequencing (RNA-seq) in both wild type (231-WT) and paclitaxel resistant type (231-PTX) of human breast cancer cell MDA-MBA-231. The two sequencing technology provide both the accurate transcript sequences and the deep transcript coverage. Then we combined shor-read and long-read RNA-seq to analyze alternative events and novel isoforms. Last but not the least, we selected BAK1 as our candidate target to verify our analysis. Our results implied that improved characterization of cancer genomic function may require the application of the single molecule long-read RNA sequencing to get the deeper and more precise view to transcriptional level. Our results imply that improved characterization of cancer genomic function may require the application of the single molecule long-read RNA sequencing to get the deeper and more precise view to transcriptional level.