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Development of a high-throughput and sensitive assay of fusion genes in lung cancer by array-based MALDI-TOFMS

机译：阵列基于Maldi-TOFM的肺癌融合基因的高通量和敏感试验

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摘要

ALK (anaplastic lymphoma kinase gene), ROS1 (ros proto-oncogene 1) and RET (ret proto-oncogene) fusions are oncogenic drivers in non-small cell lung cancer (NSCLC). Methods like fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) are highly sensitive but subjectively analyzed, labor intensive, expensive and unsuitable for multiple fusion gene screening. This study aimed to establish a high-throughput, sensitive and cost-effective screening method (array-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, array-based MALDI-TOFMS) for ALK, ROS1 and RET fusion detection. This method was established with three fusion gene positive cell lines (H2228, ALK positive; HCC78, ROS1 positive; LC-2/AD, RET positive) and negative samples. Then, 34 clinical samples were selected and detected by Sanger sequencing, next generation sequencing (NGS) and array-based MALDI-TOFMS. The results were compared and analyzed and Sanger sequencing was considered the standard. 7 cases showed ALK fusions, 1 case showed ROS1 fusions, no case showed RET fusions and 4 cases were both ALK and ROS1 fusions. Results showed that array-based MALDI-TOFMS was 100% concordant with Sanger sequencing and NGS 82.3%. In this study, we reported the utility of array-based MALDI-TOFMS in the assessment of ALK, ROS1 and RET fusions in routine lung biopsies of FFPE and fresh tissue specimens. Besides, this method may also be applied to the diagnosis, monitoring and prognosis of illness.
机译：ALK（间变性淋巴瘤激酶基因），ROS1（ROS原癌基因1）和RET（ret原癌基因）的融合体在非小细胞肺癌（NSCLC）的致癌驱动程序。像原位杂交（FISH）和免疫组织化学（IHC）的荧光的方法是高度敏感的，但主观分析，劳动密集型的，昂贵的，不适合多个融合基因的筛选。本研究旨在建立高通量，敏感和具有成本效益的筛选方法（基于阵列的基质辅助激光解吸/时间飞行电离质谱，基于阵列的MALDI-TOFMS），用于ALK，ROS1和RET融合检测。成立具有三个融合基因阳性细胞系（H2228，ALK阳性; HCC78，ROS1阳性; LC-2 / AD，RET阳性）此方法和阴性样品。然后，选择34个临床样品，通过Sanger测序，下一代测序（NGS）和基于阵列的MALDI-TOFMS进行检测。结果进行了比较和分析，Sanger测序被认为是标准。 7案件表明ALK融合，1例表现ROS1融合体，任何情况下，显示出RET融合和4例为两个ALK和ROS1融合。结果表明，基于阵列MALDI-TOFMS是100％一致的用Sanger测序和NGS 82.3％。在这项研究中，我们报道了在ALK，ROS1和RET融合在FFPE的例行肺活检和新鲜组织标本进行评估，基于阵列MALDI-TOF-MS的效用。此外，该方法也可以应用于诊断，监测和疾病的预后。

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  • 来源
    《RSC Advances》 |2018年第49期|共11页
  • 作者

    Wu Han-Tao; Li Kun; Wang Gang; Yang Xue-Xi; Zhu Anna; Xu Xu-Ping; Li Ming; Wu Ying-Song; Liu Tian-Cai;

  • 作者单位

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Guangzhou Darui Biotechnol Co Ltd Guangzhou 510665 Guangdong Peoples R China;

    Guangzhou Darui Biotechnol Co Ltd Guangzhou 510665 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

    Southern Med Univ Inst Antibody Engn Sch Lab Med &

    Biotechnol Guangzhou 510515 Guangdong Peoples R China;

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