Luo Zhanpeng, Li Dawei, Luo Xiaobo, Li Litao, Gu Suxi, Yu Long, Ma Yuanzheng
Graduate School, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Department of Orthopedics, The 309th Hospital of Chinese PLA, Beijing 100091, P.R. China.
Oncol Lett. 2018 Apr;15(4):5593-5601. doi: 10.3892/ol.2018.8032. Epub 2018 Feb 13.
The present study aimed to determine the mechanisms of action of curcumin in osteosarcoma. Human osteosarcoma U-2 OS cells was purchased from the Cell Bank of the Chinese Academy of Sciences. RNA sequencing analysis was performed for 2 curcumin-treated samples and 2 control samples using Illumina deep sequencing technology. The differentially expressed genes were identified using Cufflink software. Enrichment and protein-protein interaction network analyses were performed separately using cluster Profiler package and Cytoscape software to identify key genes. Then, the mRNA levels of key genes were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) in U-2 OS cells. Finally, cell apoptosis, proliferation, migration and invasion arrays were performed. In total, 201 DEGs were identified in the curcumin-treated group. EEF1A1 (degree=88), ATF7IP, HIF1A, SMAD7, CLTC, MCM10, ITPR1, ADAM15, WWP2 and ATP5C1, which were enriched in 'biological process', exhibited higher degrees than other genes in the PPI network. RT-qPCR demonstrated that treatment with curcumin was able to significantly increase the levels of CLTC and ITPR1 mRNA in curcumin-treated cells compared with control. In addition, targeting ITPR1 with curcumin significantly promoted apoptosis and suppressed proliferation, migration and invasion. Targeting ITPR1 via curcumin may serve an anticancer role by mediating apoptosis, proliferation, migration and invasion in U-2 OS cells.
本研究旨在确定姜黄素在骨肉瘤中的作用机制。人骨肉瘤U-2 OS细胞购自中国科学院细胞库。使用Illumina深度测序技术对2个姜黄素处理样本和2个对照样本进行RNA测序分析。使用Cufflink软件鉴定差异表达基因。分别使用cluster Profiler软件包和Cytoscape软件进行富集分析和蛋白质-蛋白质相互作用网络分析,以鉴定关键基因。然后,通过定量逆转录聚合酶链反应(RT-qPCR)检测U-2 OS细胞中关键基因的mRNA水平。最后,进行细胞凋亡、增殖、迁移和侵袭实验。总共在姜黄素处理组中鉴定出201个差异表达基因。在“生物过程”中富集的EEF1A1(度数=88)、ATF7IP、HIF1A、SMAD7、CLTC、MCM10、ITPR1、ADAM15、WWP2和ATP5C1在PPI网络中表现出比其他基因更高的度数。RT-qPCR表明,与对照组相比,姜黄素处理能够显著提高姜黄素处理细胞中CLTC和ITPR1 mRNA的水平。此外,姜黄素靶向ITPR1可显著促进细胞凋亡并抑制增殖、迁移和侵袭。姜黄素靶向ITPR1可能通过介导U-2 OS细胞的凋亡、增殖、迁移和侵袭发挥抗癌作用。