Apoptosis-Mediated Cytotoxicity of Prodigiosin-Like Red Pigment Produced by .GAMMA.-Proteobacterium and Its Multiple Bioactivities

Abstract

A bacterial strain (MS-02-063), which produces red pigment, was discovered from our library established with more than 20000 microorganisms isolated from coastal area of Nagasaki prefecture, Japan. Comparison of 16S rDNA gene sequence of strain MS-02-063 with sequences from GenBank demonstrated that this strain belongs to the g-proteobacterium species. Recent our study has demonstrated that the highly purified red pigment showed potent antimicrobial activity against pathogenic microorganisms such as Staphylococcus aureus and dermatophytes. 1)Prodigiosin (PG) analogues belonging to a family of natural red pigments are characterized by a common pyrrolydipyrrolylmethene skeleton structure with a C-4 methoxy group, and several PG-like pigments have been isolated from certain microorganisms 2-4) including some genera of actinomycetes, Pseudoalteromonas denitrificans, and Serratia marcescens. Representative members of PG analogues are prodigiosin, undecylprodigiosin, cycloprodigiosin hydrochloride, metacycloprodigiosin, and nonylprodigiosin. 5) Some of PG analogues have been reported to have several biological activities such as immunosuppressive, antimalarial, antimicrobial and cytotoxic activities. [6][7][8][9][10][11] Although the exact cytotoxic mechanisms of PG family members are not fully clarified yet, apoptotic mechanism has been proposed for some PG analogues. For instance, it has been reported that cycloprodigiosin hydrochloride induces apoptosis in cancer cells both in vitro and in vivo, with high efficiency in liver cancer, breast cancer cell lines, and colon cancer cells, [12][13][14] with no apparent toxicity to normal cells. 14) PG produced by S. marcescens also induces apoptosis in haematopoietic and gastrointestinal cancer cell lines, but no marked toxicity to non-malignant cell lines. 7,15,16) Considering the high selectivity of PG family members to cancer cells, these pigments may be promising candidates for new anticancer drugs with unique action mechanism. Regarding the structure-activity relationship, it has been reported that a nitrogen-containing heterocyclic A-pyrrole ring with the lone-pair nitrogen electrons in conjugation with the tricyclic frame and a C-4 alkoxy are important for the biological activities of PG analogues. 17)Although there are extensive studies on the cytotoxicities of PG analogues produced by S. marcescens and P. denitrificans, 4,16) only limited information on the bioactivities of PG analogues produced by g-proteobacterium is available. In this study, we examined the cytotoxic effects of PG analogue (PG-L-1), produced by g-proteobacterium (strain MS-02-063) on various cultured cell lines. Since U937 cells showed the highest susceptibility to PG-L-1 cytotoxicity, we conducted detailed analysis of cytotoxic mechanism of PG-L-1 in U937 cells especially in terms of apoptotic cell death. To search for new aspect of biochemical activity of PG-L-1, we also examined the effect of PG-L-1 on the O 2 Ϫ generation by 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated U937 cells. ...

Effects of sulfated fucan, ascophyllan, from the brown Alga Ascophyllum nodosum on various cell lines: A comparative study on ascophyllan and fucoidan

“…The colony forming efficiency of the all cell lines used was between 90% and 95%. The cytotoxicity of the polysaccharides was also assessed using the Alamar blue assay, as described previously (19). In brief, adherent cells (2500-20,000 cells/well) were cultured in 96-well plates varying concentrations of polysaccharides in the growth medium for 48 h. Then, Alamar blue reagent was added to the cells at a final concentration of 10%.…”

Mode of action of an antialgal agent produced by a marine gammaproteobacterium against Chattonella marina

“…In prelimi-nary studies, PG-L-1 led to the suppression of activation of NADPH oxidase in a macrophage cell line (data not shown). In addition, since no scavenging effect of PG-L-1 on ROS produced by hypoxanthine/xanthine oxidase system was observed (Nakashima et al 2005c), PG-L-1 may directly act on NADPH oxidaselike enzyme in C. marina. Detailed analyses of the effects of PG-L-1 on both NADPH oxidase in C. marina and macrophage cell lines are now in progress.…”

“…We suggested that the cytotoxic mechanism of PG-L-1 is the induction of apoptotic cell death through activation of p38 mitogen-activated protein kinase in human myeloid leukemia (U937) cells (Nakashima et al 2005c). Since PG-L-1 showed cytostatic activity against Chattonella marina, the cytotoxic mechanism of action of PG-L-1 against this raphidophyte may be different from that of mammalian cells.…”

“…Strain MS-02-063 was phylogenetically closely related to gammaproteobacterium MBIC 3957; however, there were differences in the physiological and biochemical properties (Nakashima et al 2005b). The red pigment produced by strain MS-02-063 had potent antifungal, antibacterial, and in vitro antitumor activities (Nakashima et al 2005a(Nakashima et al , 2005b(Nakashima et al , 2005c. Our chemical structural and biochemical analysis revealed that this red pigment belongs to the prodigiosin family, and we named it PG-L-1.…”

Pengaruh Media terhadap Produksi Prodigiosin Isolat Bakteri Entomopatogen Serratia marcescens Asal Wereng Batang Cokelat

“…Nilai ekonomis dan komersialisasi prodigiosin bersifat multifungsi, yaitu sebagai antibakteri, antifungi, antiprotozoal (Dhanasekaran et al, 2009), bersifat sitotoksik (Nakashima et al, 2005), antitumor (Castro, 1967;Pérez-Tomás et al, 2003), antimalaria, antidiabetes, antioksidan, obat-obatan, antiinflamasi non steroid, serta dapat juga menjadi pewarna untuk sutera dan wool (Khanafari et al, 2006). Produksi prodigiosin sangat bervariasi antarspesies dan strain Serratia, serta dipengaruhi oleh komposisi media dan waktu inkubasi.…”