Proteome Research of Human Lung Giant Cell Carcinoma by Two-dimensional Gel Electrophoresis and Computer Image Analysis

To optimize the two\|dimensional gel electrophoresis(2\|DE)and computer assisted image analysis for the proteome research,and to compare the proteomic difference associated with tumor metastasis,the two cell sublines C,D of PLA\|801,derived from a human lung giant cell carcinoma,were used as models.The rehydrated IPG strips containing protein samples were focuserphoresis chamber.After IEF,the equilibrated strips were transferred to the vertical SDS gel.The silver stained gels were scanned by the densitometer GS\|710,which generated the digitalized images.The computer analysis including automatic detection and quantification of protein spots,as well as alignment,matching between the different 2\|DE gels,was carried out by PDQuest software.The results demonstrated that:(1)In\|gel sample application on the IPGphor electrophoresis chamber results in more sample loading and higher resolution.(2)The IPG strips with narrow pH range got better resolution than those with wide pH range do.(3)Image analysis made it possible to globally discriminate the difference of protein profile between two sublines PLA\|801\|C and D.