Abstract
BackgroundTo better search for potential markers for hepatocellular carcinoma (HCC) invasion and metastasis, proteomic approach was applied to identify potential metastasis biomarkers associated with HCC.MethodsMembrane proteins were extracted from MHCC97L and HCCLM9 cells, with a similar genetic background and remarkably different metastasis potential, and compared by SDS-PAGE and identified by ESI-MS/MS. The results were further validated by western blot analysis, immunohistochemistry (IHC) of tumor tissues from HCCLM9- and MHCC97L-nude mice, and clinical specimens.ResultsMembrane proteins were extracted from MHCC97L and HCCLM9 cell and compared by SDS-PAGE analyses. A total of 14 differentially expressed proteins were identified by ESI-MS/MS. Coronin-1C, a promising candidate, was found to be overexpressed in HCCLM9 cells as compared with MHCC97L cells, and validated by western blot and IHC from both nude mice tumor tissues and clinical specimens. Coronin-1C level showed an abrupt upsurge when pulmonary metastasis occurred. Increasing coronin-1C expression was found in liver cancer tissues of HCCLM9-nude mice with spontaneous pulmonary metastasis. IHC study on human HCC specimens revealed that more patients in the higher coronin-1C group had overt larger tumor and more advanced stage.ConclusionsCoronin-1C could be a candidate biomarker to predict HCC invasive behavior.
Highlights
To better search for potential markers for hepatocellular carcinoma (HCC) invasion and metastasis, proteomic approach was applied to identify potential metastasis biomarkers associated with HCC
For a better insight into the characteristic of HCC metastasis, the stepwise metastatic human HCC cells MHCC97L and HCCLM9, with low and high metastatic potentials, were established via repeated in vivo selection and characterized by a similar genetic background but with significant differences in spontaneous metastasis behavior [10,11,12], providing appropriate model systems for comparative study on the molecular events correlated with HCC metastasis [13,14,15]
Differential expression of coronin-1C between HCCLM9 and MHCC97L cell strains as identified by ESI-MS/MS Membrane proteins were extracted from MHCC97L and HCCLM9 HCC cells and compared by SDS-PAGE analyses [Fig. 1A]
Summary
To better search for potential markers for hepatocellular carcinoma (HCC) invasion and metastasis, proteomic approach was applied to identify potential metastasis biomarkers associated with HCC. Most cases occur in Asia and sub-Saharan Africa [2,3], the incidence is expected to double over the 10 to 20 years in the West, possibly due to the increased HCV infection [4]. While curative therapies are possible if the lesion remains early and localized, almost 70% of resected cases recurred within 5 years [5]. The composition and characteristics of membrane proteins of tumor cells are modified during malignant transformation and make them likely candidates for cancer biomarkers [19].
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