Proteomic Characterization of Annexin l (ANX1) and Heat Shock Protein 27 (HSP27) as Biomarkers for Invasive Hepatocellular Carcinoma Cells

Abstract

To search for reliable biomarkers and drug targets for management of hepatocellular carcinoma (HCC), we performed a global proteomic analysis of a pair of HCC cell lines with distinct differentiation statuses using 2-DE coupled with MALDI-TOF MS. In total, 106 and 55 proteins were successfully identified from the total cell lysate and the cytosolic, nuclear and membrane fractions in well-differentiated (HepG2) and poorly differentiated (SK-Hep–1) HCC clonal variants, respectively. Among these proteins, nine spots corresponding to proteins differentially expressed between HCC cell types were selected and confirmed by immunofluorescence staining and western blotting. Notably, Annexin 1 (ANX1), ANX–2, vimentin and stress-associated proteins, such as GRP78, HSP75, HSC–70, protein disulfide isomerase (PDI), and heat shock protein–27 (HSP27), were exclusively up-regulated in SK-Hep–1 cells. Elevated levels of ANX–4 and antioxidant/metabolic enzymes, such as MnSOD, peroxiredoxin, NADP-dependent isocitrate dehydrogenase, α-enolase and UDP-glucose dehydrogenase, were observed in HepG2 cells. We functionally demonstrated that ANX1 and HSP27 were abundantly overexpressed only in highly invasive types of HCC cells, such as Mahlavu and SK-Hep–1. Knockdown of ANX1 or HSP27 in HCC cells resulted in a severe reduction in cell migration. The in-vitro observations of ANX1 and HSP27 expressions in HCC sample was demonstrated by immunohistochemical stains performed on HCC tissue microarrays. Poorly differentiated HCC tended to have stronger ANX1 and HSP27 expressions than well-differentiated or moderately differentiated HCC. Collectively, our findings suggest that ANX1 and HSP27 are two novel biomarkers for predicting invasive HCC phenotypes and could serve as potential treatment targets.

Discs large homolog 5 decreases formation and function of invadopodia in human hepatocellular carcinoma via Girdin and Tks5

“…D) of HCC patients. Furthermore, while high levels of Dlg5 expression were detected in HL‐7702 cells, its expression was associated negatively with invasiveness of different HCC cell lines (Fig. E).…”

Retrospective Proteomic Screening of 100 Breast Cancer Tissues

“…Mostly, they are able to bind the phospholipid of the membranes in a Ca2+ dependent manner, and to perform subsequent activities related to membrane trafficking, signal transduction, and exocytosis [63]. In our proteomics study, we have found an ubiquitous and overexpressed presence of ANX1A (3x) and of two isoform of ANXA5 (3.8x and 1.46 x respectively), which are known to play an anti-apoptotic role and are thought to be involved at different levels in the tumor development, progression and invasivity [64][65][66]. Therefore both of them are regarded as potential tumor markers [67].…”

Bradykinin stimulates protein kinase D–mediated colonic myofibroblast migration via cyclooxygenase-2 and heat shock protein 27

“…Heat shock protein 27 (Hsp27) is a molecular chaperone that is known to be involved in many cellular processes including fibrosis [15, 16], the response to stress [25], wound healing and cell migration [16, 17], and tumorigenesis [25, 26]. Hsp27 is a known downstream target of PKD [18] and is activated by post-translational phosphorylation at Serine (Ser)-15, -78, -82 and Threonine (Thr-143) residues.…”