Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University College of Medicine, Seoul, Korea.
Cancer Res Treat. 2019 Jan;51(1):391-401. doi: 10.4143/crt.2018.103. Epub 2018 Jun 14.
This study was designed to identify novel fusion transcripts (FTs) and their functional significance in colorectal cancer (CRC) lines.
We performed paired-end RNA sequencing of 28 CRC cell lines. FT candidates were identified using TopHat-fusion, ChimeraScan, and FusionMap tools and further experimental validation was conducted through reverse transcription-polymerase chain reaction and Sanger sequencing. FT was depleted in human CRC line and the effects on cell proliferation, cell migration, and cell invasion were analyzed.
One thousand three hundred eighty FT candidates were detected through bioinformatics filtering. We selected six candidate FTs, including four inter-chromosomal and two intrachromosomal FTs and each FT was found in at least one of the 28 cell lines. Moreover, when we tested 19 pairs of CRC tumor and adjacent normal tissue samples, NFATC3-PLA2G15 FT was found in two. Knockdown of NFATC3-PLA2G15 using siRNA reduced mRNA expression of epithelial-mesenchymal transition (EMT) markers such as vimentin, twist, and fibronectin and increased mesenchymal-epithelial transition markers of E-cadherin, claudin-1, and FOXC2 in colo-320 cell line harboring NFATC3-PLA2G15 FT. The NFATC3-PLA2G15 knockdown also inhibited invasion, colony formation capacity, and cell proliferation.
These results suggest that that NFATC3-PLA2G15 FTs may contribute to tumor progression by enhancing invasion by EMT and proliferation.
本研究旨在鉴定结直肠癌（CRC）细胞系中新型融合转录本（FT）及其功能意义。
我们对 28 个 CRC 细胞系进行了配对末端 RNA 测序。使用 TopHat-fusion、ChimeraScan 和 FusionMap 工具鉴定 FT 候选物，并通过逆转录聚合酶链反应和 Sanger 测序进行进一步的实验验证。在人 CRC 细胞系中敲低 FT，并分析其对细胞增殖、细胞迁移和细胞侵袭的影响。
通过生物信息学筛选检测到 1380 个 FT 候选物。我们选择了 6 个候选 FT，包括 4 个染色体间和 2 个染色体内 FT，每个 FT 至少在 28 个细胞系中的一个中被发现。此外，当我们测试 19 对 CRC 肿瘤和相邻正常组织样本时，在两个样本中发现了 NFATC3-PLA2G15 FT。使用 siRNA 敲低 NFATC3-PLA2G15 降低了上皮间质转化（EMT）标志物如波形蛋白、twist 和纤连蛋白的 mRNA 表达，并增加了 NFATC3-PLA2G15 FT 所在的 colo-320 细胞系中 E-钙黏蛋白、claudin-1 和 FOXC2 的间充质上皮转化标志物。NFATC3-PLA2G15 的敲低也抑制了侵袭、集落形成能力和细胞增殖。
这些结果表明，NFATC3-PLA2G15 FT 可能通过 EMT 增强侵袭和增殖促进肿瘤进展。
