Quantitive Detection Of Mdrl Gene And Investigation Of MDR Correlated Genes

Object To study the method of quantifying mdrl transcripts using the LightCycler PCR and its clinical application. To explore the molecular mechanism of multdrug resistance through the differential expression genes between the multdrug resistant/non-resistant ALL patiens and verify effect of some gene which function is definitude in drug resistace.Methods Appling fluorescence-quantitative reverse transcription polymerase chain reaction (RT-PCR) method of detecting mdrl gene was established and mdrl gene's expression in case of 30 leukemia and 8 healthy persons' peripheral blood were detested. Six acute lymphocyte leukemia patients were parted in resistant group and non-resistant group(three patients in each group) according mdrl gene's expression and clinic. The differentially expressed genes between the two groups were detected with BioStar 4096 cDNA microarray and were analyzed by GenePixPro 3.0 software.Cyclin B1 gene which overexpressed in resistant group was choosed,and we designed the primers according as sequence of Cyclin Bl gene and -actin gene in Genbank, Cyclin Bl gene expression level of 14 drug resistant and 13 drug non-resistant acute leukemia(AL) patients from peripheral blood were deteced by semi-quantitative reverse transcription-polymerase chain reaction.Results The standard curve of the method of detecting mdrl gene is good (The linear correlation coefficient is 1.00) . There is significant difference between the durg resistance group and the durg non-resistance group (p<0.01). Healthy persons as well contain low level mdrl transcripts. Two repeated experiment show that there were 150 differential expression genes between the multdrug resistant/non-resistant ALL patiens with cDNA microarray ,83 genes were lowexpressed and 67 genes were overexpressed,which involed in a good many aspect of cell life cycle ,for example cellgrowth,cell cycle ,cell aptosis ,ion transportion and communication conduction of cell. Semi-quantitative results of Cyclin Bl gene show that cyclin Bl gene were over expressed in drug resistant group than drug non-resistant group.Conclusion Our results indicate that fluorescence reverse transcription-polymerase chain reaction (RT-PCR) is a rapid ^ special and reliable method for quantitation of mdrl gene in clinical tumor samples, It provides a worthy parameter for clinical doctor using the medicine, preventing the MDR occurrence, monitoring conversion of the MDR . cDNA microarray has parallel and high flux aspect and offer a new means for investigation molecular pathology mechanisms of the multdrug resistant. Genes associated with cell cycle and apoptosis were abnoramlly expressed may be one of molecular pathology mechanisms of the multdrug resistant. Cyclin Bl gene can be as a maker of drug resistant of AL ,its prognostic value was suggested in AL.