| Objective To investigate the difference of glucosylceramide synthase (GCS) mRNA expression between drug-sensitive and drug-resistant human leukemia. To study the relationship between GCS and multidrug resistance in human leukemia and explore the molecular mechanism of multidrug resistance induced by GCS.Methods The clinical-dignosed leukemia peripheral blood samples were collected. After separated the plasma, the monocytes were separated by Ficoll and total RNA was abstracted. We designed the primers according as the sequence of GCS,mdr1,β-actin,bax,bcl-2 gene in Genbank. Semi-quantitative reverse transcriptional-polymerase chain reaction (RT-PCR) was employed to analyze the expression of GCS, mdr1, bcl-2 and bax mRNA in drug-sensitive and drug-resistant human leukemia. The relativity between GCS and mdr1, and between GCS and bcl-2 was analyzed separately. In additionally, ELISA was used to detect the expression of bcl-2 protein from the plasma of samples.Results 1. The comparative optical density ratios of the amplified GCS straps in the drug-resistant group are significantly higher than the drug-sensitive group (P<0.05) 2. There was the positive correlation both the expression of GCS gene and mdr1 gene in leukemia samples (P<0.01,γ= 0.66) . 3. The comparative optical density ratios of the amplified bcl-2 straps in the drug-resistant group are significantly higher than the drug-sensitive group (P<0.05), The ratio of bcl-2/bax in the drug-resistant group was higher than the drug-sensitive group(P<0.05).4. There was the positive linear correlation both the expression of GCS gene and bcl-2 gene in leukemia samples (P<0.01,γ=0.69) .5. ELISA results showed the bcl-2 protein concentration in the drug-resistant group was obviously higher than the drug-sensitive group (P<0.05) and the normal control group (P<0.01). There was positive correlation between bcl-2 protein concentration and GCS comparative optical density (γ=0.2 ) .
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