| Chemotherapy has been the important part in cancer therapy resently,and with the appliance of chemotherapeutants, the multidrug resistanceenhances increasingly. The occurrence of multidrug resistance of cancercells becomes the main factor that hinders the success of chemotherapy.Proteomics has been widespreadly implanted into every field of life scienceand medicine as an important part of post-genomics era research. Thedevelopment of theory and technology in proteomics has provided newideas and research fields for tumor multiple drug resistance. Thetwo-dimensional gel electrophoresis technology, established by O'Farrell in1975, has still been the vital technology in proteomics research dependingon its excellent resolving power. However, it has bad repetition for itsvarious processes and multifarious influences. The pH gradient formed bycarrier ampholytes drifts easily and with the occurrence of immobilized pHgradient technology, this defect has been conquered. The IPG alsoimproves the 2-DE's repetition and resolving power infinitely.Two-dimensional gel electrophoresis was used with immobilized pHgradient to display and compare the differential expression ofmultidrug-resistant proteins in gastric cancer cells SGC7901 andSGC7901/ADR, in order to find the new multidrug-resistant protein andmake the foundation for further studying the gastric carcinoma multidrugresistance mechanism. Methods: The total proteins of SGC7901 and SGC7901/ADR wereextracted by chemistry lysis method and Bradford method was used todetected their concentration. The total proteins of the two cell lines wereseparated by immobilized pH-gradient-based 2-DE and the spots ofproteins were visualized by silver staining. The differentially expressedproteins were analyzed using ImagingMaster 2D Melanie 5.0 software. Results: 1.the protein extracting rate 106 can be extracted 191ug~230ug protein averagely. 2.the feature of the 2-DE image 381 and 354 protein spots were identified in the 2-DE gel of SGC7901and SGC7901/ADR individually and the matched spots were 259. Theseprotein spots are within the range of pI3.3~9.8 and 14.4kD~133.0kD andthere were 233 spots range in pI4~8 and 30kD~97kD in SGC7901 cells,while there were 186 spots in that area in SGC7901/ADR cells. Nineproteins were seen to be unique high in one region or the others (4 inSGC7901/ADR cells, 5 in SGC7901 cells). There were 85 over expressionspots in the SGC7901/ADR cells, and there were 46 protein spots(>2,<5fold) ranged in pI4.0~9.8, 14.4kD~89.4kD, 20 protein spots(>5,<10 fold)ranged in pI3.3~9.5, 14.6kD~65.6kD, 19 protein spots(>10 fold) ranged inpI4.9~9.8, 14.3kD~95.4kD. There were 75 over expression spots in theSGC7901 cells, and there were 44 protein spots(>2,<5 fold) ranged inpI4.1~9.7, 16.7kD~126.3kD, 17 protein spots(>5,<10 fold) ranged inpI3.4~9.5, 14.8kD~64.9kD, 14 protein spots(>10 fold) ranged in pI5.3~9.2,22.3kD~65.4kD. Conclusion: The whole-cell proteins of gastric cancer cells SGC7901 andSGC7901/ADR could be well separated and displayed by two-dimensionalelectrophoresis. The results suggest that these differential proteins berelated to the Adriamycin–resistant mechanism in human gastric cancer cellline SGC7901/ADR.
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