科研之友
微信
新浪微博
Facebook
分享链接
摘要
OBJECTIVE To investigate the effects of theaflavin-3,3′-digallate (TF3) on proliferation and apoptosis of human osteosarcoma HOS cells and to explore its underlying mechanism through experiments including transcriptomics. METHODS Human osteosarcoma cells were treated with different concentrations of TF3 and 0.05% DMSO, respectively. Then, cell proliferation was detected by CCK-8 assay, crystal violet staining, colony formation assay and flow cytometry; cell apoptosis rate was detected by Hoechst33258 staining and flow cytometry. After treatment with of TF3 at different concentrations, the protein levels of the molecules related to proliferation and apoptosis of osteosarcoma HOS cells were determined by Western blot. The potential molecular mechanism of TF3 on HOS cells in vitro was revealed by transcriptomics and bioinformatic analysis. Real-time fluorescent quantitative PCR was performed in order to verify SCG2, EPHB1, and CLDN7 mRNA expression levels in cells based on results of transcriptome sequencing. RESULTS Compared with the control group, TF3 inhibited proliferation of osteosarcoma HOS cells and promoted apoptosis (P＜0.05) in a concentration-dependent manner. The expression levels of proliferation related proteins PCNA and Ki67 were all down-regulated by TF3 (P＜0. 01). TF3 decreased the expression of antiapoptotic protein Bcl-2 and apoptosis-related protein Caspase-3 (P＜0. 01). TF3 promoted the expression of apoptotic markers Bax, cleaved caspase-3 and cytochrome C (P＜0.01). Transcriptome analysis revealed 809 differential expressed genes(DEGs) after TF3 treatment in HOS cells, including 213 down-regulated and 596 up-regulated genes. The gene ontology (GO) analysis showed that the function of DEGs were mainly enriched in positive regulation of DNA replication, membrane, protein binding, etc. The Kyoto encyclopedia of genes and genome (KEGG) analysis showed that the function of DEGs were mainly enriched in apoptosis and tumor-related signaling pathways, including pathways in cancer, HIF-1 signaling pathway, TNF signaling pathway, etc. The expression of SCG2, EPHB1, and CLDN7 declined in comparison with the control group (P＜0.05). CONCLUSION TF3 inhibits the proliferation of osteosarcoma HOS cells, and also promote its apoptosis. Its molecular mechanism is potentially related to the regulation of multi-gene and multi-signaling pathway.
- 单位
