Abstract
Hepatocellular carcinoma (HCC) is currently the sixth most common malignancy and the second major cause of tumor-related deaths in the world. This study aimed to investigate the role of cleavage and polyadenylation factor-6 (CPSF6) and B-cell translocation gene 2 (BTG2) in regulating the glycolysis and apoptosis in HCC cells. The RNA and protein expression of CPSF6 and BTG2 in normal hepatocyte and HCC were, respectively, detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis and Western blot analysis. The viability and apoptosis of transfected Huh-7 cells were, respectively, analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. The expression of apoptosis-related proteins and HK-2 in transfected Huh-7 cells was also detected by Western blot analysis. The levels of glucose and lactate in the culture supernatant of transfected Huh-7 cells were, respectively, detected with the glucose assay kit and lactate assay kit. The interaction of CPSF6 and BTG2 was confirmed by RNA binding protein immunoprecipitation (RIP) assay. As a result, CPSF6 expression was increased while BTG2 expression was decreased in Huh-7 cells. Interference with CPSF6 suppressed the viability and glycolysis, and promoted the apoptosis of Huh-7 cells. Furthermore, CPSF6 interacted with BTG2 and interference with CPSF6 upregulated the BTG2 expression and inhibited the protein kinase B (AKT)/extracellular signal-regulated kinase (ERK)/nuclear factor (NF)-κB pathway. Interference with BTG2 could partially reverse the above cell changes caused by interference with CPSF6. In conclusion, CPSF6 inhibited the BTG2 expression to promote glycolysis and suppress apoptosis in HCC cells by activating AKT/ERK/NF-κB pathway.
Highlights
Hepatocellular carcinoma (HCC) was a pathological type with the highest incidence in liver malignant tumor, accounting for about 85% of primary liver cancer
HER2overexpressing and triple-negative subtypes depend on CPSF6 for viability and tumorigenic capacity in aggressive breast cancer cells of luminal B, and CPSF6 is overexpressed in human breast cancer cases, which is associated with poor prognosis [7]
Expression level of CPSF6 in HCC cells The mRNA expression of CPSF6 was obviously upregulated in hepatocellular carcinoma cells compared with THLE-3 cells and the mRNA expression of CPSF6 in Huh-7 cells was the highest (Fig. 1A)
Summary
Hepatocellular carcinoma (HCC) was a pathological type with the highest incidence in liver malignant tumor, accounting for about 85% of primary liver cancer. The incidence of HCC was the sixth highest among all malignancies and the second highest among all tumor-associated causes of death [1, 2]. Despite the improvement and optimization of HCC treatment protocols in recent years, the poor prognosis of HCC patients remains unchanged. HER2overexpressing and triple-negative subtypes depend on CPSF6 for viability and tumorigenic capacity in aggressive breast cancer cells of luminal B, and CPSF6 is overexpressed in human breast cancer cases, which is associated with poor prognosis [7]. ENCORI (http://starbase.sysu.edu.cn/index.php) shows that CPSF6 is upregulated in patients with HCC and is associated with poor prognosis (Additional file 1: Figure S1A, B)
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