Effect Of Gemcitabine On Granulocyte Colony-stimulating Factor Receptor And Bcr/abl Mrna About Chronic Myelogenous Leukemia

Objective:With the advance of molecular biology,the nucleoside analogues have become a group of antitumor agents which have been used widely in the treatment of hematological malignancies.Gemcitabine(GEM) is an anti-metabolism anticancer drug.Gemcitabine is not easy deaminated due to its special structure which hydrogens at the 2'-carbon are replaced by geminal fluorine atoms.Simultaneously,Gemcitabine has more water-solubility because of hydrochloric acid's addition at pyrimidine position.As a new kind of deoxycytidine analogue,Gemcitabine has strong cytotoxicity and unique self-enhancement mechanism.At present,it has been widely applied in the treatment of patients with solid tumors.It's therapeutic effect was identificated and side reactions could be tolerated.So the FDA approved gemcitabine as the first line chemotherapeutic drug in the treatment of non-small cell lung cancer, pancreatic cancer and metastatic breast cancer.In overseas,the research of gemcitabine in treatment of leukemia is still in the stage of fundamental researches and clinical trials.In domestic,gemcitabine in treatment of refractory and relapsed lymphoma have been reported.But the reports on gemcitabine in the treatment of leukemia are rare.Graunlocyte colony-stimulating factor(G-CSF)is considerable hemopoietic growth factor.By combining together the effector cell surface acceptor (G-CSF receptor,G-GSFR),G-CSF can regulate granulocytic multiplication and differentiation.In this experiment,G-GSFR and bcr/abl mRNA of the CML cells were observed after cells treated with gemcitabine.Furthermore,the possibility of gemcitabine to treat leukemia was discussed.Methods:1.Twenty-three cases of CML chronic phase,eight cases of CML blast crisis and ten cases of normal control were involved in the study.We obtained the marrow 6ml from these cases.The bone marrow mononucleus cell(MNC) liquid were made to cultured for 48h in different concentration of GEM(0μg/ml,10μg/ml).2.The mRNA expressions of BCR/ABL was assessed by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR).The expressions of G-CSFR were detected by flow cytometry.3.The data were analyzed using software SPSS 10.0.The numeration data were analyzed with the x~2 test.Data were presented as mean±standardifference. The multi-samples mean values were compared by variance analysis.Correlation test:analysis of Pearson correlation.P values of less than 0.05 was considerd statistecally significant.Results:1.After the MNC were treated with 0μg/ml GEM for 48h,the expression rates of G-CSFR were(45.42±6.52)%,(32.58±5.68)%,and(58.56±5.54)%in CML chronic phase,CML blastic phase and normal control respectively.After the MNC were treated with 10μg/ml GEM for 48h,the expression rates of G-CSFR were (50.72±8.57)%,(36.32±4.25)%,and(59.42±7.62)%in CML chronic phase, CML blastic phase and normal control respectively.After cultured with 10μg/ml gemcitabin for 48h,the expression rates of G-CSFR in CML chronic phase and CML blastic phase were significantly different than that with 0μg/ml GEM for 48h (P＜0.05).2.After the MNC cultured with 0μg/ml GEM for 48h,the expression of BCR/ABL mRNA cells growth were 0.60±0.10 and 0.63±0.11 in CML chronic phase and CML blastic phase.After the MNC cultured with 10μg/ml GEM for 48h,the expression of BCR/ABL mRNA cells growth were 0.59±0.15 and 0.60±0.13 in CML chronic phase and CML blastic phase.There was no significant difference between 0μg/ml GEM group and 10μg/ml GEM group(P＞0.05).There is a negative relationship between mRNA expreesion of BCR/ABL,and the expression rates of G-CSFR of CML chronic phase and CML blastic phase in bone marrow.Conclusions:1.The expression rates of G-CSFR of CML chronic phase and CML blastic phase are significantly lower than that of normal control.2.The expression rates of G-CSFR of CML chronic phase are significantly higher than that of CML blastic phase.3.There is a negative relationship between mRNA expreesion of BCR/ABL,and the expression rates of G-CSFR of CML chronic phase and CML blastic phase in bone marrow.4.The expression rates of G-CSFR in CML chronic phase and CML blastic phase were raised by gemcitabin in vitro culture.