Plasmacytoma inhibitory factors secreted by marcophage induced apoptosis

• Main Authors: Kang-Sing Wang, 王康馨
• Other Authors: Jerming tseng
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/37474211992396092979

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.