Bao Le, Yuan Lei, Li Pengfei, Bu Qingyun, Guo Aijun, Zhang Hui, Cui Ning, Liu Bin
Department of Intervention, Xuzhou Cancer Hospital, Xuzhou Hospital Affiliated to Jiangsu University, Xuzhou, China.
Department of Intervention, Xuzhou City Hospital of TCM, Xuzhou, China.
Cell Physiol Biochem. 2018;50(2):437-451. doi: 10.1159/000494155. Epub 2018 Oct 11.
BACKGROUND/AIMS: The roles and related mechanisms of RNA binding protein FUS (fused in sarcoma/translocated in liposarcoma) are unclear in numerous cancers, including hepatocellular carcinoma (HCC).
Quantitative reverse transcription PCR (qRT-PCR), western blot, cell viability, transwell migration and invasion, tumor spheres formation and in vivo tumor formation assays were used to examine the effects of FUS on HCC progression in HuH7 and MHCC97 cells. Additionally, transcriptome analysis based on RNA-sequencing data, qRT-PCR, western blots, luciferase reporter and RNA binding protein immunoprecipitation (RIP) assays were used to explore the LATS1/2 (large tumor suppressor kinases 1/2)-related mechanisms contributing to FUS functions. Finally, qRT-PCR and western blot analysis were used to detect the levels of FUS and LATS1/2 in HCC and adjacent normal tissues, and the correlation between them in HCC tissues.
Overexpression of FUS decreased cell viability, migration, invasion and stemness. Moreover, FUS interacted and stabilized LATS1/2 stability, and thus promoted LATS1/2 expression and activated Hippo pathway. Finally, FUS and LAST1/2 levels were positively correlated and significantly down-regulated in HCC tissues.
We demonstrate that FUS/LATS1/2 axis inhibits HCC progression via activating Hippo pathway.
背景/目的：在包括肝细胞癌（HCC）在内的多种癌症中，RNA结合蛋白FUS（肉瘤融合蛋白/脂肪肉瘤易位蛋白）的作用及其相关机制尚不清楚。
采用定量逆转录PCR（qRT-PCR）、蛋白质免疫印迹法、细胞活力检测、Transwell迁移和侵袭实验、肿瘤球形成实验以及体内肿瘤形成实验，研究FUS对HuH7和MHCC97细胞中HCC进展的影响。此外，基于RNA测序数据的转录组分析、qRT-PCR、蛋白质免疫印迹法、荧光素酶报告基因实验以及RNA结合蛋白免疫沉淀（RIP）实验，用于探索与FUS功能相关的LATS1/2（大肿瘤抑制激酶1/2）相关机制。最后，通过qRT-PCR和蛋白质免疫印迹分析检测HCC及癌旁正常组织中FUS和LATS1/2的水平，以及它们在HCC组织中的相关性。
FUS过表达降低细胞活力、迁移、侵袭和干性。此外，FUS相互作用并稳定LATS1/2的稳定性，从而促进LATS1/2表达并激活Hippo信号通路。最后，FUS和LAST1/2水平呈正相关，且在HCC组织中显著下调。
我们证明FUS/LATS1/2轴通过激活Hippo信号通路抑制HCC进展。
