miR-193a-3p Promotes the Invasion, Migration, and Mesenchymal Transition in Glioma through Regulating BTRC.

BACKGROUND: The present study is aimed at exploring the specific expression of miR-193a-3p and the mechanism underlying miR-193a-3p-mediated mesenchymal transition (MT), invasion, and migration in glioma. METHODS: The gene expression profile datasets of GSE39486 and GSE25676 were downloaded from the National Center for Biotechnology (NCBI). Data regarding the expression of miR-193a-3p and survival curves were derived from Chinese Glioma Genome Atlas (CGGA). Online websites including miRWalk, DIANA, and starbase were employed to predict the target genes for miR-193a-3p. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed by the Omicsbean online software. Module analysis of the protein-protein interaction (PPI) networks was performed by the plug-in Molecular Complex Detection (MCODE), and the degrees of genes were calculated by CytoHubba plug-in of Cytoscape. Survival curves were based on the Gene Expression Profile Interaction Analysis (GEPIA). Transwell, wound healing, and Western blot experiments were performed to investigate the effects of miR-193a-3p and beta-transducin repeat containing E3 ubiquitin protein ligase (BTRC) on the invasion, migration, and MT of glioma. RESULTS: miR-193a-3p was highly expressed in glioma tissues and significantly correlated with poor survival in patients with glioma. The target genes for miR-193a-3p were involved in many cancer-related signaling pathways. The PPI showed 11 genes with both high degrees and MCODE scores in the network. Survival analysis demonstrated that the expression of BTRC was significantly correlated with the prognosis of patients with glioma. The results from the transwell, wound healing, and Western blot analyses suggested that miR-193a-3p promoted the invasion, migration, and MT of glioma cells, which could be reversed by BTRC. CONCLUSIONS: miR-193a-3p was upregulated in patients with glioma and could affect the invasion, migration, and MT of glioma by regulating BTRC.

Chromodomain Helicase/ATPase DNA-Binding Protein 1-Like Gene (CHD1L) Expression and Implications for Invasion and Metastasis of Breast Cancer.

BACKGROUND: Chromodomain helicase/ATPase DNA-binding protein 1-like gene (CHD1L), also known as ALC1 (amplified in liver cancer 1 gene), is a new oncogene amplified in many solid tumors. Whether this gene plays a role in invasion and metastasis of breast cancer is unknown. METHODS: Immunohistochemistry was performed to detect the expression of CHD1L in patients with invasive ductal carcinoma and normal mammary glands. Chemotaxis, wound healing, and Transwell invasion assays were also performed to examine cell migration and invasion. Western blot analysis was conducted to detect the expression of CHD1L, MMP-2, MMP-9, pAkt/Akt, pARK5/ARK5, and pmTOR/mTOR. Moreover, ELISA was carried out to detect the expression levels of MMP-2 and MMP-9. Nude mice xenograft model was used to detect the invasion and metastasis of breast cancer cell lines. RESULTS: CHD1L overexpression was observed in 112 of 268 patients (41.8%). This overexpression was associated with lymph node metastasis (P = 0.008), tumor differentiation (P = 0.020), distant metastasis (P = 0.026), MMP-2 (P = 0.035), and MMP-9 expression (P = 0.022). In the cell experiment, reduction of CHD1L inhibited the invasion and metastasis of breast cancer cells by mediating MMP-2 and MMP-9 expression. CHD1L knockdown via siRNA suppressed EGF-induced pAkt, pARK5, and pmTOR. This knockdown inhibited the metastasis of breast cancer cells into the lungs of SCID mice. CONCLUSIONS: CHD1L promoted the invasion and metastasis of breast cancer cells via the PI3K/Akt/ARK5/mTOR/MMP signaling pathway. This study identified CHD1L as a potential anti-metastasis target for therapeutic intervention in breast cancer.

[Effect of 5-Aza-dC on FHIT gene expression in hepatocellular carcinoma cell line HepG2].

BACKGROUND AND OBJECTIVE: FHIT gene methylation leads to down-regulation of its expression in hepatocellular carcinoma (HCC) cells. This study was to detect the expression of FHIT mRNA and protein in HCC cell line HepG2 after treatment of methylase inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC), and observe the effect of 5-Aza-dC on the proliferation of HepG2 cells. METHODS: HepG2 cells were treated with 5-Aza-dC. Methylation of FHIT in HepG2 cells was detected by methylation-specific polymerase chain reaction (MSP). FHIT mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). FHIT protein expression was detected by immunohistochemistry and Western blot. Cell proliferation was detected by MTT assay. RESULTS: Before treatment of 5-Aza-dC, FHIT gene methylation was detected in HepG2 cells, while no FHIT mRNA and protein expression was detected. The hypermethylation of FHIT gene in HepG2 cells was effectively reversed after treatment of 5-Aza-dC. When HepG2 cells were treated with 1.0, 2.0, and 4.0 micromol/L of 5-Aza-dC for 48 h, the mRNA levels of FHIT were 0.80+/-0.32, 1.41+/-0.54, and 1.51+/-0.61, respectively; the protein levels of FHIT were 0.33+/-0.20, 1.00+/-0.26, and 1.12+/-0.38, respectively. Cell proliferation was significantly inhibited after being treated with 5-Aza-dC. CONCLUSION: 5-Aza-dC can reverse the abnormal methylation of FHIT gene, activate the silenced gene and induce FHIT mRNA and protein expression in HepG2 cells.