SELDI-TOF-MS Technology In Analyzing The Trace Protein Chart In Nude Mouse Transplanted Tumor Model Of Liver Cancer

Objective: To build the nude mouse transplanted tumor modelsof human liver cancer by using Bel-7402human liver cancer cells suspension tovaccinate age of3weeks’ nude mouse in hypoderm of left shoulder and back,and observe the relationship between the tumor size and growth time,measuring and analyzing the differentially expressed proteins of nude mouse indifferent times after the animal modle is built successfully by usingSELDI-TOF-MS technology, analyzing the expression pattern of these traceproteins, aimed at finding new diagnostic markers of the primary carcinoma ofliver in early stage. Methods: Recovering Bel-7402human liver cancer cells,choosing the cells in logarithmic growth phase to configure cells suspension of1×107/ml after subcultured3-5times, vaccinating each nude mouse of the3experimental groups in left shoulder and back’s hypoderm by0.1ml,vaccinating nude mouse of control group with equivalent serum free medium atthe same place, taking close observation in the growth of tumor of theexperimental animals, measuring and recording the tumor size of nude mouseevery two weeks. Collecting all the nude mice’s blood samples by excisingtheir eye balls, collect0.5ml blood samples each mouse of control group and3experimental groups separately at20d、40d、60d after the animal model is builtsuccessfully, then put nude mouse to death, separate the tumor tissue and sendit to the department of pathology to do the biopsy. The blood samples of nudemouse should be centrifuged for5min at4℃with3000rpm at once toseparate the blood serum, split charging and mark numbers on the serumsamples, then keep them at-80℃. Analyzing the trace protein in serum samplesof nude mouse with SELDI-TOF-MS technology, collecting and analyzing thedata of the protein spectra we get with the software Ciphergen ProteinChip3.0 and Ciphergen Biomaker Wizard3.1, screening out the differentially expressedproteins that associated with liver cancer, searching and paralleling them in theprotein database. Results: The animal model is built successfully after15-20d,the tumor formation rate is100%. There are5differentially expressed proteins(P<0.01) that associated with liver cancer and changed regularly we get, whoserelative molecular mass are2016Da、3309Da、3442Da、3745Da、2747Da,the first four trace proteins’ expression are gradually increased in theexperimental group nude mouse, and the last race protein’s expression isgradually attenuated in the experimental group nude mouse. Searching these5trace proteins in Swiss-Prot protein database, the preliminary results are Protein2A、Potassium channel toxin alpha-KTx20.1、Serine protease inhibitor2、Neurotoxin P2and LeuA leader peptide. Conclusion:1.The method that buildanimal tumor model by vaccinating nude mouse with Bel-7402human livercancer cells suspension in hypoderm has advantages such as easy to operate、time is short、tumor formation rate is high, provides an effective method for thescientific research of liver cancer.2. Analyzing the differentially expressedproteins of nude mouse with SELDI-TOF-MS technology, probably can findserum markers for diagnosis of liver cancer, open up a new way for the study ofearly diagnosis of liver cancer.