ABSTRACT
<p><b>OBJECTIVE</b>To establish a multidrug-resistant hepatocellular carcinoma cell line BEL-7402/5-deoxy-5-fluorouridine (5'-DFUR) and study the mechanisms of the drug resistance.</p><p><b>METHODS</b>BEL-7402/5'-DFUR cell line was induced by pulse therapy combined with continuous stepwise exposure to 5'-DFUR in vitro. The multidrug resistance of BEL-7402/5'-DFUR cell line to the antitumor agents was evaluated by MTT assay. The distribution of the cell cycle, the expressions of P-gp, bcl-2 and GST-pi were detected by flow cytometry.</p><p><b>RESULTS</b>The established BEL-7402/5'-DFUR cell line was resistant to multiple antitumor agents, with IC(50) of 5'-DFUR 12.9 times higher than that of the parental cell line 7204. The BEL-7402/5'-DFUR cells in S phase decreased while those in G(1) and G(2) phase increased, with significantly increased expressions of P-gp and bcl-2 but stable expression of GST-pi.</p><p><b>CONCLUSION</b>Compared with its parent cell line BEL-7402, the multidrug resistant cell line BEL-7402/5'-DFUR has a 12.9-fold increase in IC(50) of 5'-DFUR with decreased drug accumulation and altered cell cycle distribution. The multi-drug resistance of this cell line is closely related to the overexpression of P-gp and bcl-2.</p>
Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Antimetabolites, Antineoplastic , Pharmacology , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Line, Tumor , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Floxuridine , Pharmacology , Glutathione Transferase , Metabolism , Liver Neoplasms , Metabolism , Pathology , Proto-Oncogene Proteins c-bcl-2 , MetabolismABSTRACT
In order to investigate the clinical efficacy of non-myeloablative allogeneic stem cell transplantation (allo-NST) and related technology in patients with hematologic malignancies, twenty-six cases of hematological malignancies (10 AL, 14 CML, 2 MM patients) received NST following conditioning regimens with fludara + cyclophosphamide + ATG (14 cases) and busulfan or melphalan + cyclophosphamide + ATG (12 cases), G-CSF (600 micro g/d) or G-CSF (300 micro g/d) + GM-CSF (300 micro g/d) were used for mobilizing peripheral blood stem cell. A combination of cyclosporine A (CsA) and methotrexate (MTX) was administered for GVHD prophylaxis. Patients will be eligible for donor lymphocyte infusion (DLI) or donor stem cell infusion (DSI) given in graded increments according to the chimeric formation and clinical reaction. Generally the dose of the first infusion was 1 x 10(7)/kg at 4th week post-transplantation. The engraftment analysis included the detection of microsatellite short tandem repeats (STRs), Bcr/Abl fusion gene, Philadelphia chromosome, HLA-locus analysis, sex chromosome and ABO blood type or blood subtype. The results showed that 22 patients (84.62%) were engrafted, among which 18 patients were full donor chimerism (FDC) up to now. Acute GVHD occurred in 3/26 cases (11.54%). Chronic GVHD was diagnosed in 6 of 26 (23.07%) evaluable patients. The incidence of infection and hemorrhage was low and slight. It is concluded that allo-NST is a safe and effective therapeutic method for hematologic malignancies, but the related technology such as selection of indication, conditioning regimen and transplantation immunotherapy should be studied further.