Overexpression of CENP-H as a novel prognostic biomarker for human hepatocellular carcinoma progression and patient survival

Abstract

Centromere protein H (CENP-H) has been shown to be significantly upregulated in many types of cancers and is associated with disrupted cell cycle regulation, cell proliferation and genetic instability. The aim of the present study was to explore the expression and localization of CENP-H in hepatocellular carcinoma (HCC) and determine whether its overexpression is a prognostic biomarker for HCC. Reverse transcription-polymerase chain reaction (pcr), real-time qPCR and western blotting were used to compare CENP-H expression at the mRNA and protein levels in HCC samples and corresponding adjacent non-cancerous samples. CENP-H protein levels were determined in 60 paired paraffin-embedded HCC tissues using immunohistochemistry (IHC), and the correlation with clinicopathological features and patient prognosis was analyzed. In addition, an immunofluorescence assay was performed to test the expression and localization of CENP-H protein in HCC cells. Results showed that levels of CENP-H mRNA and protein were higher in HCC samples than in the corresponding adjacent non-cancerous samples. In 60 paired paraffin-embedded tissues, CENP-H was upregulated in the HCC samples (38/60, 63.3%) relative to the adjacent non-cancerous samples (21/60, 35%, P=0.003), and a higher level of upregulation was associated with tumor size (P=0.032); higher histological grade (P=0.001); more advanced TNM stage (P=0.002) and Chinese clinical stage (P=0.008); and poorer prognosis. In addition, consistent with the results of IHC, the immunofluorescence assay showed that CENP-H was localized in the nucleus of Hep3B cells. CENP-H was overexpressed in HCC, and its level of upregulation was an independent prognostic indicator, suggesting that CENP-H may be an effective therapeutic strategy for the treatment of HCC.

CENP-H regulates the cell growth of human hepatocellular carcinoma cells through the mitochondrial apoptotic pathway

“…In our previous research, we found that CENP-H expression is not only upregulated in HCC tissues at both the mRNA and protein levels but is also closely related to tumor size, histological grade, and clinical stage. Additionally, CENP-H expression based on immunohistochemistry was negatively associated with patient prognosis (15). In the present study we further confirmed that the expression levels of CENP-H were higher in the HCC cell lines than that in the L02 immortalized human liver cells.…”

“…Total cell protein was extracted according to a previous study (15). After denaturation, equal amounts of total protein were separated electrophoretically on 10% SDS/polyacrylamide gels (SDS-PAGE) and transferred onto polyvinylidene difluoride (PvDF) membranes (Millipore, Bedford, MA, USA) using a tank transfer apparatus (Bio-Rad Laboratories).…”

“…Xenograft samples were immediately fixed in 4% formalin once obtained from the mice. After paraffin-embedding, 4-µm thick paraffin sections were adhered to glass slides and prepared for immunohistochemical assay using a standardized streptavidin-peroxidase (SP) method as previously reported (15). Following deparaffinization, rehydration, antigen retrieval and blockage, the slides were incubated with primary antibodies overnight at 4̊C.…”

“…Moreover, it may be a novel prognostic biomarker of HCC progression (15). In the present study, we investigated the role of CENP-H in HCC cell survival.…”

TXNRD1 Is an Unfavorable Prognostic Factor for Patients with Hepatocellular Carcinoma

“…IHC was performed according to the standard method described previously [12, 13]. Two observers who were blinded to the clinical outcome evaluated and scored the degree of immunostaining of TXNRD1 independently.…”