Abstract
Background and AimPrognostic markers are important for predicting the progression and staging of hepatocellular carcinoma (HCC). Ezrin (EZR) and Podocalyxin (PODXL) are proteins associated with invasion, migration and poor prognosis in various types of cancer. Recently, it has been observed that chloride intracellular channel 5 (CLIC5) forms a complex with EZR and PODXL and that it is required for podocyte structure and function. In this study, we evaluated the overexpression of EZR, PODXL and CLIC5 in HCC.MethodsThe modified resistant hepatocyte model (MRHR), human biopsies and HCC cell lines (HepG2, Huh7 and SNU387) were used in this study. Gene and protein expression levels were evaluated in the MRHR by qRT-PCR, Western blot and immunohistochemistry analyses, and protein expression in the human biopsies was evaluated by immunohistochemistry. Protein expression in the HCC cell lines was evaluated by immunofluorescence and Western blot, also the migration and invasive abilities of Huh7 cells were evaluated using shRNA-mediated inhibition.ResultsOur results indicated that these genes and proteins were overexpressed in HCC. Moreover, when the expression of CLIC5 and PODXL was inhibited in Huh7 cells, we observed decreased migration and invasion.ConclusionThis study suggested that EZR, CLIC5 and PODXL could be biological markers to predict the prognosis of HCC and that these proteins participate in migration and invasion processes.
Highlights
Invasion is a critical step in the development of metastasis, which is characterized by changes in the expression of proteins related to the cytoskeleton, migration, and invasion [1]
It has been observed that chloride intracellular channel 5 (CLIC5) forms a complex with EZR and PODXL and that it is required for podocyte structure and function
This study suggested that EZR, Chloride intracellular channel 5 (CLIC5) and PODXL could be biological markers to predict the prognosis of hepatocellular carcinoma (HCC) and that these proteins participate in migration and invasion processes
Summary
Invasion is a critical step in the development of metastasis, which is characterized by changes in the expression of proteins related to the cytoskeleton, migration, and invasion [1]. Several cell membrane and cytoskeleton-related proteins are overexpressed in many types of cancers, including Ezrin (EZR) [2] and Podocalyxin (PODXL) [3], which are associated with morphogenesis and migration because they function as scaffold proteins between the cell membrane and actin cytoskeleton [4, 5]. We examined the expression of EZR, CLIC5 and PODXL at the mRNA and protein levels in a modified hepatocyte resistant model (MHRM) and in cases of human hepatocellular carcinoma (HCC). Ezrin (EZR) and Podocalyxin (PODXL) are proteins associated with invasion, migration and poor prognosis in various types of cancer. It has been observed that chloride intracellular channel 5 (CLIC5) forms a complex with EZR and PODXL and that it is required for podocyte structure and function. We evaluated the overexpression of EZR, PODXL and CLIC5 in HCC
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