| Summary: | 碩士 === 國立師範大學 === 生物學研究所 === 82 === Inhibitory factors secreted by P388D1 macrophage-like cell
line exerted both inhibitory effect and cytotoxic effect on MOPC-
315 plasmacytoma. The inhibitory factors were distinctfrom the de
fined cytotoxic factors, such as neutral protease,interleukin-1,
interleukin-6, tumor necrosis factor andarginase. In this study,
the inhibitory factors in P388D1 supernatant were purified using
DEAE-Sephacel ion-exchange chromatography. The major proteins
were separated into five groups. Proteins in peak 4 and peak 5
showed inhibitory effect on the growth and IgA secretion of MOPC-
315. SDS-PAGE analysis of the five groups of proteins showed that
peak 4 contained 102.3-KDa and 17.9-KDa proteins which had been
observed by others using Sephacryl S-300 chromatography analysis
in previous report. Peak 5 consisted of a low molecular weight pr
otein which may be the monomer component of high molecular weight
inhibitory factor reported by other. Furthermore, we demonstrated
that plasmacytoma inhibitory factors induced the programmed cell
death (apoptosis) of MOPC-315 cell. In the early stage of apoptos
is,the endonucleases were activated and the DNA molecules were cl
eaved into fragments that showed mutiples of 180-200 base pairs.
DNA fragments could be visulized on gel electrophoresis and the
amount of fragmented DNA was dose dependent. The DPA reaction mod
ified from Burton provided an additional evidence ofapoptosis.
The result of DPA assay showed that P388D1 supernatant generated
DNA fragmentation in adose-dependentmanner and the amount of DNA
fragments reached aplateau at 72 hours. Peak 4 and peak 5 obtain-
ing from DEAE-Sephacel also increase the amount of DNA fragments.
In addition, the fractions with higher inhibition activity genera
ted more DNA fragmentation.
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