title: Reversion of malignant phenotypes of human glioblastoma cells by β-elemene through β-catenin-mediated regulation of stemness-, differentiation- and epithelial-to-mesenchymal transition-related molecules.
abstract: BACKGROUND
  Glioblastoma is the most common and lethal type of primary brain tumor. β-Elemene, a natural plant drug extracted from Curcuma wenyujin, has shown strong anti-tumor effects in various tumors with low toxicity. However, the effects of β-elemene on malignant phenotypes of human glioblastoma cells remain to be elucidated. Here we evaluated the effects of β-elemene on cell proliferation, survival, stemness, differentiation and the epithelial-to-mesenchymal transition (EMT) in vitro and in vivo, and investigated the mechanisms underlying these effects.
  
  
  METHODS
  Human primary and U87 glioblastoma cells were treated with β-elemene, cell viability was measured using a cell counting kit-8 assay, and treated cells were evaluated by flow cytometry. Western blot analysis was carried out to determine the expression levels of stemness markers, differentiation-related molecules and EMT-related effectors. Transwell assays were performed to further determine EMT of glioblastoma cells. To evaluate the effect of β-elemene on glioblastoma in vivo, we subcutaneously injected glioblastoma cells into the flank of nude mice and then intraperitoneally injected NaCl or β-elemene. The tumor xenograft volumes were measured every 3 days and the expression of stemness-, differentiation- and EMT-related effectors was determined by Western blot assays in xenografts.
  
  
  RESULTS
  β-Elemene inhibited proliferation, promoted apoptosis, impaired invasiveness in glioblastoma cells and suppressed the growth of animal xenografts. The expression levels of the stemness markers CD133 and ATP-binding cassette subfamily G member 2 as well as the mesenchymal markers N-cadherin and β-catenin were significantly downregulated, whereas the expression levels of the differentiation-related effectors glial fibrillary acidic protein, Notch1, and sonic hedgehog as well as the epithelial marker E-cadherin were upregulated by β-elemene in vitro and in vivo. Interestingly, the expression of vimentin was increased by β-elemene in vitro; this result was opposite that for the in vivo procedure. Inhibiting β-catenin enhanced the anti-proliferative, EMT-inhibitory and specific marker expression-regulatory effects of β-elemene.
  
  
  CONCLUSIONS
  β-Elemene reversed malignant phenotypes of human glioblastoma cells through β-catenin-involved regulation of stemness-, differentiation- and EMT-related molecules. β-Elemene represents a potentially valuable agent for glioblastoma therapy.
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. zhutingzhun2000@aliyun.com.
authors.forename: Tingzhun
authors.initials: T
authors.lastname: Zhu
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. lixm1376212743@aliyun.com.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Li
authors.affiliation: Health Care Centre, Shenyang Entry-Exit Inspection and Quarantine Bureau, Shenyang, China. luolihan258@aliyun.com.
authors.forename: Lihan
authors.initials: L
authors.lastname: Luo
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. littlepig@hotmail.com.
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. lizhiqing@yeah.net.
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. pengxie308@yahoo.com.
authors.forename: Peng
authors.initials: P
authors.lastname: Xie
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. gaoelmer@hotmail.com.
authors.forename: Xu
authors.initials: X
authors.lastname: Gao
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. szqlz@163.com.
authors.forename: Zhenquan
authors.initials: Z
authors.lastname: Song
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. jingyuansu2014@yahoo.com.
authors.forename: Jingyuan
authors.initials: J
authors.lastname: Su
authors.affiliation: Department of Neurosurgery, General Hospital of Shenyang Military Area Command, No. 83, Wenhua Road, Shenhe District, Shenyang, 110840, China. syjqzyyliangguobiao@yahoo.com.
authors.forename: Guobiao
authors.initials: G
authors.lastname: Liang
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology*; D064593:Heterografts; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology; D012717:Sesquiterpenes / Q000494:pharmacology*; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D012717:Sesquiterpenes; D051176:beta Catenin; C445979:beta-elemene