Abstract | BACKGROUND: METHODS: Cell viability was assessed using the CCK-8 assay. Cell cycle analysis was performed using flow cytometry. Apoptosis was evaluated using Annexin V staining and flow cytometry, Hoechst 33342 staining and the TUNEL assay. Expression and cleavage of caspase-3, caspase-9 and poly ADP-ribose polymerase (PARP) were assessed by Western blotting. Statistical analysis was performed using two-tailed Student's t-tests. The gene expression profiles of U251 glioma cells treated with HATi II or DMSO were analyzed using the Arraystar Human 8 x 60 K LncRNA/ mRNA expression array; data was analyzed using MEV (Multi Experiment View) cluster software. Datasets representing genes with altered expression profiles (≥2-fold) derived from the cluster analyses were subjected to gene ontology and pathway analysis. RESULTS: HATi II inhibited the proliferation of U251, U87, HS683 and SHG44 cells in a dose-dependent manner. HATi II induced cell cycle arrest at the G2/M phase, and induced significant levels of apoptosis, apoptotic body formation and DNA fragmentation in HATi II-treated U251 and SHG44 cells. HATi II induced cleavage of caspase-3, caspase-9 and PARP in U251 and SHG44 cells. In HATi II-treated U251 cells, 965 genes were upregulated, 984 genes were downregulated and 3492/33327 lncRNAs were differentially expressed. GO analysis showed the differentially expressed genes with known functions are involved in a variety of processes; alcoholism, p53 signaling pathway, cytokine- cytokine receptor interaction and transcriptional mis-regulation in cancer were the four most significant pathways. Upregulation of p53 signaling pathway-related genes in HATi II-treated cells was confirmed by quantitative RT-PCR and Western blotting. CONCLUSIONS: HATi II inhibits proliferation and induces apoptosis via the caspase-dependent pathway in human glioma cell lines, possibly by activating the p53 signaling pathway. HATi II deserves further investigation as a novel treatment for glioma.
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Authors | Li-Xiao Xu, Zhi-Heng Li, Yan-Fang Tao, Rong-Hu Li, Fang Fang, He Zhao, Gang Li, Yan-Hong Li, Jian Wang, Xing Feng, Jian Pan |
Journal | Journal of experimental & clinical cancer research : CR
(J Exp Clin Cancer Res)
Vol. 33
Pg. 108
(Dec 19 2014)
ISSN: 1756-9966 [Electronic] England |
PMID | 25523932
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Apoptosis Regulatory Proteins
- Histone Deacetylase Inhibitors
- RNA, Messenger
- TP53 protein, human
- Tumor Suppressor Protein p53
- HDAC2 protein, human
- Histone Deacetylase 2
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Topics |
- Apoptosis
(drug effects)
- Apoptosis Regulatory Proteins
(metabolism)
- Brain Neoplasms
(drug therapy, enzymology, genetics, pathology)
- Cell Cycle
(drug effects)
- Cell Line, Tumor
- Cell Proliferation
(drug effects)
- Cell Survival
(drug effects)
- Dose-Response Relationship, Drug
- Gene Expression Profiling
(methods)
- Gene Expression Regulation, Neoplastic
- Glioma
(drug therapy, enzymology, genetics, pathology)
- Histone Deacetylase 2
(antagonists & inhibitors, metabolism)
- Histone Deacetylase Inhibitors
(pharmacology)
- Humans
- Oligonucleotide Array Sequence Analysis
- RNA, Messenger
(metabolism)
- Real-Time Polymerase Chain Reaction
- Signal Transduction
(drug effects)
- Time Factors
- Tumor Suppressor Protein p53
(metabolism)
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