Polo-like kinase 1, a new therapeutic target in hepatocellular carcinoma
| Autor: | Wei Chuen Mok, Seng Gee Lim, Shanthi Wasser, Theresa M.C. Tan |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Carcinoma Hepatocellular Blotting Western Fluorescent Antibody Technique Mice Nude Apoptosis Cell Cycle Proteins Biology Protein Serine-Threonine Kinases Real-Time Polymerase Chain Reaction Transfection Amino Acid Chloromethyl Ketones Mice Proto-Oncogene Proteins Gene expression Animals Humans Aged Cell Proliferation Aged 80 and over Gene knockdown TUNEL assay Endodeoxyribonucleases Cell growth Caspase 3 Reverse Transcriptase Polymerase Chain Reaction Liver Neoplasms Gastroenterology General Medicine Genetic Therapy Hep G2 Cells Middle Aged Molecular biology Caspase Inhibitors Xenograft Model Antitumor Assays digestive system diseases Tumor Burden enzymes and coenzymes (carbohydrates) Terminal deoxynucleotidyl transferase Tumor progression Cell culture Gene Knockdown Techniques Cancer research Original Article Female RNA Interference biological phenomena cell phenomena and immunity |
| Popis: | AIM: To investigate the role of polo-like kinase 1 (PLK1) as a therapeutic target for hepatocellular carcinoma (HCC). METHODS: PLK1 gene expression was evaluated in HCC tissue and HCC cell lines. Gene knockdown with short-interfering RNA (siRNA) was used to study PLK1 gene and protein expression using real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, and cell proliferation using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2(4-sulfophenyl)-2H-tetrazolium (MTS) and bromodeoxyuridine (BrdU) assays. Apoptosis was evaluated using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and caspase-inhibition assay. Huh-7 cells were transplanted into nude mice and co-cultured with PLK1 siRNA or control siRNA, and tumor progression was compared with controls. RESULTS: RT-PCR showed that PLK1 was overexpressed 12-fold in tumor samples compared with controls, and also was overexpressed in Huh-7 cells. siRNA against PLK1 showed a reduction in PLK1 gene and protein expression of up to 96% in Huh-7 cells, and a reduction in cell proliferation by 68% and 92% in MTS and BrdU cell proliferation assays, respectively. There was a 3-fold increase in apoptosis events, and TUNEL staining and caspase-3 assays suggested that this was caspase-independent. The pan-caspase inhibitor Z-VAD-FMK was unable to rescue the apoptotic cells. Immnofluorescence co-localized endonuclease-G to fragmented chromosomes, implicating it in apoptosis. Huh-7 cells transplanted subcutaneously into nude mice showed tumor regression in siPLK1-treated mice, but not in controls. CONCLUSION: Knockdown of PLK1 overexpression in HCC was shown to be a potential therapeutic target, leading to apoptosis through the endonuclease-G pathway. |
| Databáze: | OpenAIRE |
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