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Identification of differentially expressed proteins in tumour necrosis factor-alpha-resistant and -sensitive rat
Yasuhiro Kuramitsu1, Xiulian Zhang, Yufeng Wang
1Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan. climates@yamaguchi-u.ac.jp
Background:
Our earlier studies reported that ONO-4007, a synthetic lipid A analogue with low endotoxic activity, had shown much effect on tumour necrosis factor (TNF)-alpha-sensitive rat hepatoma cells, even though it had no effect on TNF-alpha-resistant cells.
Materials And Methods:
To find biomarkers which relate to the sensitivity of cancer cells to TNF-alpha, proteomic differential display analysis for TNF-alpha-resistant cKDH-8/11 and -sensitive KDH-8/YK rat hepatoma cell lines was carried out using two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry.
Results:
Two-DE analysis showed 32 spots, whose expression was different between cKDH-8/11 cells and KDH-8/YK cells. Of these, 12 were up-regulated and 20 were down-regulated in cKDH-8/11 cells compared to KDH-8/YK cells. The up-regulated proteins include transitional endoplasmic reticulum ATPase, 78 kDa glucose-regulated protein (GRP78), heat-shock cognate 71 kDa protein (HSC71) and protein disulfide-isomerase A6. The down-regulated proteins included alpha-enolase, aldose reductase, glutathione reductase, annexin A1, glutamate dehydrogenase 1 and dihydrolipoyl dehydrogenase.
Conclusion:
These findings suggest that these differentially regulated proteins could be factors responsible for the resistance of cKDH-8/11 cells to TNF-alpha-induced cell death.
Insights
Researchers identified protein biomarkers linked to cancer cell resistance to tumor necrosis factor-alpha (TNF-α). Differential proteomic analysis revealed specific proteins that may explain why some cancer cells survive TNF-α treatment.
Area of Science:
- Cancer Biology
- Proteomics
- Cellular Signaling
Background:
- Previous research indicated ONO-4007, a lipid A analog, affects tumor necrosis factor-alpha (TNF-α)-sensitive hepatoma cells but not resistant ones.
- This highlights a need to understand the molecular mechanisms of TNF-α resistance in cancer.
Purpose of the Study:
- To identify protein biomarkers associated with cancer cell sensitivity or resistance to TNF-α.
- To elucidate the proteomic differences between TNF-α-sensitive and TNF-α-resistant rat hepatoma cell lines.
Main Methods:
- Proteomic differential display analysis was performed on TNF-α-resistant (cKDH-8/11) and TNF-α-sensitive (KDH-8/YK) rat hepatoma cell lines.
- Techniques included two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Main Results:
- Differential 2-DE analysis identified 32 protein spots with altered expression between the two cell lines.
- Twelve proteins were upregulated in resistant cells, including GRP78 and HSC71.
- Twenty proteins were downregulated in resistant cells, including alpha-enolase and annexin A1.
Conclusions:
- The identified differentially regulated proteins are potential biomarkers for TNF-α resistance.
- These proteins may play a role in the resistance of cKDH-8/11 cells to TNF-α-induced apoptosis.
