Proliferation Ability Of HepG2 Cells Research Articles

Profiles of Acetylation Regulation Genes Contribute to Malignant Progression and Have a Clinical Prognostic Impact on Liver Cancer.

Background Several studies have demonstrated that acetylation was involved in the process of liver cancer. This study aimed to establish an effective predictive prognostic model using acetylation regulation genes in liver cancer. Methods Two datasets were downloaded from the Cancer Genome Atlas (TCGA) database and International Cancer Genome Consortium (ICGC) database. Differentially expressed acetylation regulation genes were identified in the TCGA-LIHC dataset, and then, Gene Ontology (GO) functional annotation analysis was used to investigate the molecular mechanism. After grouping the patients into clusters based on consensus clustering, we explored the correlation between clusters and clinical characteristics. A risk model was constructed by the least absolute shrinkage and selection operator (LASSO) regression analysis to calculate the risk score. Patients were divided into high-risk and low-risk groups according to the risk score using the acetylation regulation genes. Data downloaded from LIRI-JP were used for external validation. Univariate and multivariate Cox regressions were performed to identify independent risk factors. A prognostic nomogram was constructed according to the TCGA-LIHC dataset. The effect of HDAC11 expression on the proliferation and migration of liver cancer was detected by the CCK-8 method and cell scratch test, respectively. Results Eleven of 29 acetylation regulation genes were identified as upregulated differentially expressed genes. Go enrichment analysis showed that they were involved in “protein and histone deacylation and deacetylation.” Patients were categorized into two clusters according to the expression of 29 acetylation regulation genes. Compared with cluster 2, cluster 1 correlated with shorter overall survival (OS) and higher expression. Stage, T stage, grade, gender, age, and follow-up state were significantly different between two clusters. Pathways involved in DNA repair were significantly enriched in cluster 1. The risk score was calculated by HDAC1, HDAC2, HDAC4, HDAC11, HAT1, and SIRT6. Patients in the high-risk group had a worse prognosis in both datasets. Risk score was not only an independent prognostic marker but could also predict the clinicopathological features of liver cancer. A nomogram containing risk score, T stage, and M stage was built to predict overall survival. After transfection with HDAC11 overexpression plasmid, the proliferation ability of HepG2 cells increased, while the migration ability had no change. Conclusions Our findings suggested that acetylation regulation genes contribute to malignant progression and have a clinical prognostic impact on liver cancer.

FOXP3 expression in FOXP3+ tumor cells promotes hepatocellular cells metastasis.

BackgroundForkhead transcription factor 3 (FOXP3) is a key molecule for the development of regulatory T cell. Recent studies showed that FOXP3 was also expressed in tumor cells. This study is designed to identify the expression of FOXP3 and its pathogenesis in hepatocellular carcinoma (HCC).MethodsThrough immunohistochemistry, RNA extraction and Real-Time Quantitative PCR, western blot analysis, transwell cell migration assay and invasion assays, and in vivo experiment, we detected FOXP3 expression in HCC and analyzed the expression of tumor metastasis-related genes in HCC cells using female BALB/c-Nude mice.ResultsThe results showed that FOXP3 was expressed in partial HCC tissues samples and cell lines. The distant metastasis rate was remarkably higher in the FOXP3 positive HCCs than that in the negative group. The positive rate of FOXP3 expression in the metastatic HCC was higher than that in primary HCC, and the expression level of FOXP3 was found to increase as the enhancement in the metastatic potential of the cell lines. Furthermore, in HCC cell lines, FOXP3 overexpression can promote the cell metastasis and invasion by regulating MMP-1. The in vivo experiment showed that the proliferation ability of HepG2 cells in nude mice increased significantly after FOXP3 was overexpressed, and the incidence rate of lung metastasis in MHCC97L cells was significantly deceased after knocking down the FOXP3 expression.ConclusionsOur findings support that partial HCC tissues and cell lines expressing FOXP3 can promote the metastasis by regulating MMP-1.

Up-regulation of CXCL8 expression is associated with a poor prognosis and enhances tumor cell malignant behaviors in liver cancer

CXCL8, a member of CXC chemokines, was constitutively expressed in many types of human cancers, and its overexpression has been shown to play a critical role in promoting tumorigenesis. The purpose of the present study was to determine CXCL8 expression in a commercial human liver tissue microarray, and elucidate the effects and underlying mechanisms by which CXCL8 is involved in the malignant progression of human liver cancer. Our data showed that high level expression of CXCL8 in tissues with liver cancer was identified as compared with non-cancer tissues, and its up-regulation was closely associated with clinical stage and tumor infiltration. In vitro, exogenous CXCL8 at concentrations of 10, 20 or 40 ng/ml obviously stimulated the proliferation abilities of HepG2 cells. Coupled with this, 10, 20 or 40 ng/ml of exogenous CXCL8 also triggered a significant elevation in HepG2 cells migration. Additionally, overexpression of CXCL8 in HepG2 cells also resulted in increased cell proliferation and migration capacities. Finally, Western blotting analysis showed that overexpression of CXCL8 increased the expression of ERK, p-ERK and survivin, decreased the expression of caspase-3 and BAX at protein level.

The effects of microRNA-7 on proliferation and invasion of hepatocellular carcinoma HepG2 cells

Objective: To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro. Methods: The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3'UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis. Results: The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05). Conclusions: The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.