Effects of tetrazanbigen on the protein expression in human hepatocellular carcinoma cell line QGY-7701

Abstract

Tetrazanbigen (TNBG) is a novel synthetic antitumor drug with significant antitumor effects on common solid tumors in vitro and in vivo. It may lead to death of cancer cells through a tumor-associated lipoidosis mechanism, and result in lipid droplets (LDs) accumulation at the cytoplasm. In this study, the effects of TNBG on protein expression in human hepatocellular carcinoma cell line QGY-7701 were studied for elucidating its antitumor mechanism. The proteins extracted from TNBG-treated human hepatocellular carcinoma cell line QGY-7701 were analyzed and compared with control cells by two-dimensional gel electrophoresis. The differential proteins were identified by matrix-associated laser desorption ionization time-of-flight mass (MALDI-TOF-MS) spectrometry. Two proteins of interest, the levels of which were significantly increased in TNBG-treated cells, were further characterized by Western blot analysis. The results showed a total of 846±23 spots in control cells and 853±30 spots in TNBG-treated cells. Twenty-six up-regulated or down-regulated proteins were found by analyzing differential proteomic 2-DE map. Eleven of them were identified by mass spectrometry. They were protein disulfide-isomerase precursor, 94 kD glucose-regulated protein, heat shock protein (HSP) 90-alpha, ATP-citrate lyase, HMG-CoA reductase, glucose-6-phosphate 1-dehydrogenase, very-long-chain specific acyl-CoA dehydrogenase, squalene synthetase, sterol regulatory element-binding protein 1, fructose-bisphosphate aldolase A, and peroxiredoxin-1. These up-regulated or down-regulated proteins are mostly related to lipid metabolism. The TNBG antitumor mechanism is probably to influence tumor lipid metabolism, resulting in accumulation of LDs in tumor cells.

Keywords

tetrazanbigen / antitumor / lipid metabolism / proteomics

References

[1] Yu Y. Preparation and use of a kind of azagonane compound. Chinese patent (Chinese), 1996. CN1124251

[2] LiL.J., YuY., TangW.X., et al. . Effects of TNBG on the proliferation and apoptosis of human hepatocellular carcinoma cell line QGY-7701. J Sichuan Univ (Med Sci) (Chinese), 2005, 36: 372-374

[3] LiL.J., YangX.L., YuanY.H., et al. . Changes of gene expression of human hepatocellular carcinoma cell line QGY-7701 induced by tetrazanbigen. Acta Academiae Medicinae Militaris Tertiae (Chinese), 2006, 28: 151-153

[4] HwangY.Y., LiM.D.. Protein differentially expressed in response to nicotine in five rat brain regions: Identification using a 2-DE/MS-based proteomics approach. Proteomics, 2006, 6: 3138-3153

[5] JiangD., YingW., LuY., et al. . Identification of metastasis-associated proteins by proteomic analysis and functional exploration of interleukin-18 in metastasis. Proteomics, 2003, 3: 724-737

[6] RamagliL.S., RodriguezL.V.. Quantitation of microgram amounts of protein in two-dimensional polyacrylamide gel electrophoresis sample buffe. Electrophoresis, 1985, 6: 559-563

[7] BergquistJ., GobomJ., BlombergA., et al. . Identification of nuclei associated proteins by 2D-gel electrophoresis and mass pectrometry. J Neurosci Meth, 2001, 109: 3-11

[8] NeuhoffV., AroldN., TaubeD., et al. . Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitity using Coomassie brilliant blue G-250 and R-250. Electrophoresis, 1988, 9: 255-262

[9] GernerC., FrohweinU., GotzmannJ., et al. . The Fas-induced apoptosis analyzed by high throughput proteome analysis. J Biol Chem, 2000, 275(50): 39018-39026

[10] PageM.J., AmessB., RohlffC., et al. . Proteomics: A major new technology for the drug discovery process. Drug Discov Today, 1999, 4: 55-62

[11] de CromR., van HaperenR., JanssensR., et al. . Grp96/Grp94 is a putative high density lipoprotein-binding protein in liver. Biochim Biophys Acta, 1999, 1437(3): 378-392

[12] LanH., RabagliaM.E., SchuelerK.L., et al. . Distinguishing covariation from causation in diabetes:a lesson from the protein disulfide isomerase mRNA abundance trait. Diabetes, 2004, 53: 240-244

[13] WetterauJ.R., CombsK.A., McleanL. e. a1.. Protein disulfide isomerase appears necessary to maintain the catalytically active structure of the microsomal triglyceride transfer protein. Biochemistry, 1991, 30(40): 9728-9735

[14] EberleD., HegartyB., BossardP. e. a1.. SREBP transcription factors: master regulators of lipid homeostasis. Biochimie, 2004, 86(11): 839-848

[15] CcombsG., WestropG., SuehanP. e. a1.. The Amitechondriate eukaryote triehomonas vaginalis contains a divergent thioredoxin-1inked peroxiredoxin antioxidant system. J Bio Chem, 2004, 279(7): 5249-5256

[16] BeyerN.H., RoepstorfP., HammerK. e. a1.. Protcome analysis of the purine stimulon from Lactecaceus lactis. Proteomics, 2003, 3(5): 786-797

[17] LiB.H., MaX.F., WangY., et al. . Structure-activity relationship of polyphenols that inhibit Fatty Acid Synthase. J Biochem (Tokyo), 2005, 138: 679-685

[18] ScagliaN., CavigliaJ.M., IgalR.A.. High stearoyl-CoA desaturase protein and activity levels in simian virus 40 transformed-human lung fibroblasts. Biochim Biophys Acta, 2005, 1687: 141-151

[19] GoldsteinJ.L., BrownM.S.. Regulation of the mevalonate pathway. Nature, 1990, 343: 425-430

[20] JakobisiakM., GolabJ.. Potential antitumor effects of statins (Review). Int J Oncol, 2003, 23: 1055-1069