Disruption of endocytic trafficking protein Rab7 impairs invasiveness of cholangiocarcinoma cells
| Autor: | Sopit Wongkham, Nantana Suwandittakul, Onrapak Reamtong, Santi Maneewatchararangsri, Maleerat Sutherat, Urai Chaisri, Poom Adisakwattana, Pattamaporn Molee |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cancer Research information science Biology Immunofluorescence Cholangiocarcinoma 03 medical and health sciences 0302 clinical medicine Downregulation and upregulation Cell Line Tumor parasitic diseases Genetics medicine Humans Neoplasm Invasiveness cardiovascular diseases Matrigel Gene knockdown medicine.diagnostic_test fungi rab7 GTP-Binding Proteins General Medicine Prognosis Subcellular localization Blot 030104 developmental biology Bile Duct Neoplasms Oncology rab GTP-Binding Proteins Cell culture Cytoplasm 030220 oncology & carcinogenesis cardiovascular system Cancer research |
| Zdroj: | Cancer Biomarkers. 20:255-266 |
| ISSN: | 1875-8592 1574-0153 |
| DOI: | 10.3233/cbm-170030 |
| Popis: | Background Alterations and mutations of endo-lysosomal trafficking proteins have been associated with cancer progression. Identification and characterization of endo-lysosomal trafficking proteins in invasive cholangiocarcinoma (CCA) cells may benefit prognosis and drug design for CCA. Objective To identify and characterize endo-lysosomal trafficking proteins in invasive CCA. Methods A lysosomal-enriched fraction was isolated from a TNF-α induced invasive CCA cell line (KKU-100) and uninduced control cells and protein identification was performed with nano-LC MS/MS. Novel lysosomal proteins that were upregulated in invasive CCA cells were validated by real-time RT-PCR. We selected Rab7 for further studies of protein level using western blotting and subcellular localization using immunofluorescence. The role of Rab7 in CCA invasion was determined by siRNA gene knockdown and matrigel transwell assay. Results Rab7 mRNA and protein were upregulated in invasive CCA cells compared with non-treated controls. Immunofluorescence studies demonstrated that Rab7 was expressed predominantly in invasive CCA cells and was localized in the cytoplasm and lysosomes. Suppression of Rab7 translation significantly inhibited TNF-α-induced cell invasion compared to non-treated control (p= 0.044). Conclusions Overexpression of Rab7 in CCA cells was associated with cell invasion, supporting Rab7 as a novel candidate for the development of diagnostic and therapeutic strategies for CCA. |
| Databáze: | OpenAIRE |
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