Upregulation of heat shock protein 70 and the differential protein expression induced by tumor necrosis factor-alpha enhances migration and inhibits apoptosis of hepatocellular carcinoma cell HepG2

Abstract

Tumor necrosis factor alpha (TNFα) plays diverse roles in liver damage and hepatocarcinogenesis with its multipotent bioactivity. However, the influence of TNFα on protein expression of hepatocellular carcinoma (HCC) is incompletely understood. Therefore, we aimed to investigate the differential protein expression of HCC in response to TNFα stimulus. We observed that HepG2 cell revealed a higher resistance to TNFα-induced apoptosis as compared to the non-tumorigenic hepatocyte THLE-2. By using a label-free quantitative proteomic analysis, we found that 520 proteins were differentially expressed in the HepG2 cells exposed to TNFα, including 211 up-regulated and 309 down-regulated proteins. We further confirmed several proteins with significant expression change (TNFα/control ratio>2.0 or <0.5) by immunoblotting using specific antibodies. We also analyzed the differential expressed proteins using Gene ontology and KEGG annotations, and the results implicated that TNFα might regulate ribosome, spliceosome, antigen processing and presentation, and energy metabolism in HepG2 cells. Moreover, we demonstrated that upregulation of heat shock protein 70 (HSP70) was involved in both the promoted migration and the inhibited apoptosis of HepG2 cells in response to TNFα. Collectively, these findings indicate that TNFα alters protein expression such as HSP70, which triggering specific molecular processes and signaling cascades that promote migration and inhibit apoptosis of HepG2 cells.

Bioinformatics analysis of the interactions among lncRNA, miRNA and mRNA expression, genetic mutations and epigenetic modifications in hepatocellular carcinoma

“…Therefore, investigating a single factor associated with gene regulation is not sufficient to assess the prognosis of patients with HCC. Previous studies have identified genes that regulate HCC progression; however, the majority of these studies focused on single genes or investigated only one of the factors influencing gene regulation (16)(17)(18), without considering their combinatorial effect. To understand the complex molecular biological characteristics of HCC, differentially expressed lncRNAs, miRNAs and mRNAs were identified by analyzing biological datasets from TCGA database.…”

Bergapten induces G1 arrest and pro‐apoptotic cascade in colorectal cancer cells associating with p53/p21/PTEN axis

“…Cell viability was assessed by MTT assay as previously described . Briefly, cells were seeded at the initial density of 2 × 10 5 cells/well in a 24‐well plate, cultured with serum‐free DMEM for 16 hours, and then incubated with serial concentrations of bergapten (10, 30, and 50 μM) for 24 or 48 hours.…”

“…Crude proteins (20 μg of protein) were subjected to a 12.5% SDS‐polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane as previously described . The blot was blocked using 5% nonfat milk in PBS, incubated with a primary antibody against specific proteins, and then the bound primary antibodies were detected by peroxidase‐conjugated secondary antibody and ECL chemiluminescence developing reagent.…”

HspA1BIs a Prognostic Biomarker and Correlated With Immune Infiltrates in different subtypes of Breast Cancers

“…HSP70 is regulated by the cell cycle and is kept in a low level in normal cells. But in tumor cells, the expression level is increased due to the stimulation by mutant or abnormal proteins [61]. Some research showed that in tumor microenvironment, Hsp70 /Hsp70 complex (HSP70 /HSP70-PCs) plays a complex and important roles.…”