Τίτλος:
mRNA expression analysis of a variety of apoptosis-related genes,
including the novel gene of the BCL2-family, BCL2L12, in HL-60 leukemia
cells after treatment with carboplatin and doxorubicin
Περίληψη:
Apoptosis is a type of programmed cell death involved in many crucial
biological processes. It represents the basic mechanism for the action
of chemotherapeutic agents, such as doxorubicin and carboplatin. Both
are able to cause cell death through the induction of apoptosis in the
human leukemic cell line HL-60. We investigated the possible alterations
in the expression of apoptosis-related genes, including the novel
BCL2L12 gene, which was recently cloned in our group. The kinetics of
apoptosis induction and cell toxicity was investigated by DNA laddering
and by the MTT method, respectively. Total RNA was extracted and cDNA
was prepared by reverse transcription. BCL2, BAX, FAS, caspase-9,
caspase-3 and BCL2L12 were amplified by PCR. Overexpression of FAS,
BCL2L12 and caspase-3 was observed after treatment of HL-60 cells for 3
or 6 In with carboplatin, while their expression was decreased after a
12-h treatment, demonstrating that these genes may take part in the
early stages of apoptosis. Overexpression of the same genes was also
observed after 6 h of treatment with doxorubicin (concomitantly with DNA
laddering). In the case of carboplatin-induced apoptosis we detected
down-regulation of BAX, BCL2 and caspase-9, whereas in the case of
doxorubicin, BAX and BCL2 remained at control levels and caspase-9 was
increased.
Συγγραφείς:
Floros, KV
Thomadaki, H
Katsaros, N
Talieri, M
Scorilas,
A