Effect of bleomycin and cisplatin on the expression profile of SRA1, a novel member of pre-mRNA splicing factors, in HL-60 human promyelocytic leukemia cells.
Recently, a new member of the human SR (Ser/Arg-rich) superfamily of pre-mRNA splicing factors, SRA1 (SR-A1), has been discovered and cloned by members of our group, the gene for which was found to be overexpessed in a series of human tumors. In the present study, we investigated the significance of alterations at the mRNA expression levels of the SRA1 gene after treatment of HL-60 human promyelocytic leukemia cells with the anticancer drugs cisplatin and bleomycin. The kinetics of apoptosis and cell toxicity were investigated by DNA laddering and the MTT and trypan blue assays, respectively. Total RNA was extracted and cDNA was prepared by reverse transcription. The splicing-related genes SRA1 and SC35, as well as the apoptosis-related gene BCL2 (Bcl-2), were amplified by PCR using gene-specific primers. The results showed that mRNA levels of SRA1 were up-regulated upon treatment with the antibiotic bleomycin, whereas they were down-regulated by treatment of HL-60 human promyelocytic leukemia cells with cisplatin. Our results support the hypothesis that mRNA expression analysis of SRA1 may serve as a new prospective molecular marker, playing an important role in chemotherapy outcome in human leukemia.
Katsarou
Maria E
ME
National Center for Scientific Research Demokritos, IPC, GR-15310 Athens, Greece.
Thomadaki
Hellinida
H
Katsaros
Nikos
N
Scorilas
Andreas
A
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001761:Bleomycin / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D053938:DNA Fragmentation / Q000187:drug effects; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics*; D012322:RNA Precursors / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000235:genetics*; D000068103:Serine-Arginine Splicing Factors; D013997:Time Factors
D000970:Antineoplastic Agents; D009687:Nuclear Proteins; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D001761:Bleomycin; D000068103:Serine-Arginine Splicing Factors; D002945:Cisplatin