Gong Xubo, Wang Baiyong, Yan Lijuan, Lu Xiaoya, Zhao Xiaoying
Department of Clinical Laboratory, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zheijang 310000, P.R. China.
Department of Intensive Care Unit, The First Hospital of Hangzhou Normal University, Hangzhou, Zheijang 310000, P.R. China.
Oncol Lett. 2020 Nov;20(5):181. doi: 10.3892/ol.2020.12042. Epub 2020 Aug 31.
Linalool can inhibit the malignant proliferation of numerous human malignant solid tumors, including hepatocellular carcinoma, breast cancer, small cell carcinoma and malignant melanoma. However, the role of linalool in T cell acute lymphoblastic leukaemia (T-ALL) remains unclear. In the present study, human T-ALL cell lines (Jurkat, H9, Molt-4 and Raji cells) and peripheral blood mononuclear cells (PBMCs) from healthy donors were treated with various concentrations of linalool (3.75, 7.50, 15.00, 30.00, 60.00 and 120.00 µM, respectively). A CCK-8 assay was used to analyse cell viability and it demonstrated that linalool inhibited the growth of T-ALL cells in a dose-dependent manner, but did not significantly affect normal PBMCs. Flow cytometry was used to detect the cell cycle and apoptosis and demonstrated that linalool reduced the percentage of T-ALL cells at the G/G phase, and induced the apoptosis of T-ALL cells. RNA sequencing was conducted on an Illumina HiSeq X Series 2500 before and after treatment with linalool followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. It was demonstrated that the mitogen-activated protein kinase (MAPK) pathway was involved in the effect of linalool on T-ALL cells. Real-time quantitative PCR and western blotting were performed to verify the mRNA and protein levels, respectively of the genes in the signaling pathway identified. In addition, it was found that linalool significantly inhibited phosphorylated (p)-ERK1/2 protein expression and enhanced p-JNK protein expression of T-ALL cells. In conclusion, the present study revealed that linalool inhibits T-ALL cell survival with involvement of the MAPK signaling pathway. JNK activation and ERK inhibition may play a functional role in apoptosis induction of T-ALL cells. Linalool may be developed as a novel anti T-ALL agent.
芳樟醇能够抑制多种人类恶性实体瘤的恶性增殖，包括肝细胞癌、乳腺癌、小细胞癌和恶性黑色素瘤。然而，芳樟醇在T细胞急性淋巴细胞白血病（T-ALL）中的作用仍不清楚。在本研究中，用不同浓度的芳樟醇（分别为3.75、7.50、15.00、30.00、60.00和120.00 μM）处理人T-ALL细胞系（Jurkat、H9、Molt-4和Raji细胞）以及健康供体的外周血单个核细胞（PBMC）。采用CCK-8法分析细胞活力，结果表明芳樟醇以剂量依赖性方式抑制T-ALL细胞的生长，但对正常PBMC没有显著影响。使用流式细胞术检测细胞周期和凋亡情况，结果表明芳樟醇降低了处于G/G期的T-ALL细胞百分比，并诱导了T-ALL细胞的凋亡。在用芳樟醇处理前后，在Illumina HiSeq X Series 2500上进行RNA测序，随后进行基因本体论和京都基因与基因组百科全书通路富集分析。结果表明，丝裂原活化蛋白激酶（MAPK）通路参与了芳樟醇对T-ALL细胞的作用。进行实时定量PCR和蛋白质印迹分别验证所鉴定信号通路中基因的mRNA和蛋白质水平。此外，发现芳樟醇显著抑制T-ALL细胞中磷酸化（p）-ERK1/2蛋白的表达，并增强p-JNK蛋白的表达。总之，本研究表明芳樟醇通过MAPK信号通路抑制T-ALL细胞存活。JNK激活和ERK抑制可能在T-ALL细胞凋亡诱导中发挥作用。芳樟醇可能被开发为一种新型抗T-ALL药物。
