Molt 4B Cells Research Articles

A lymphocyte-generated fragment of vasoactive intestinal peptide with VPAC1 agonist activity and VPAC2 antagonist effects.

Vasoactive intestinal peptide receptors 1 (VPAC1) and 2 (VPAC2) have been identified in humans. Cell lines expressing only VPAC1 (HT-29) or VPAC2 (Molt-4b) were identified using real-time reverse transcriptase polymerase chain reaction. Vasoactive intestinal peptide (VIP) and related peptides, VIP-6-28, VIP4-28, and VIP10-28, previously isolated from cultures of human leukocytes, were evaluated for their ability to bind to VPAC1 and VPAC2 and to increase the levels of cAMP in HT-29 and Molt-4b cells. VIP bound to membranes of HT-29 colon carcinoma cells and Molt-4b lymphoblasts with high affinity (KD = 1.6 +/- 0.2 and 1.7 +/- 0.9 nM, respectively). VIP4-28 also demonstrated high-affinity binding (KD = 1.7 +/- 0.2 and 1.7 +/- 0.7 nM in HT-29 and Molt-4b, respectively). VIP and VIP4-28 are potent VPAC1 agonists, inducing maximal 200- and 400-fold increases in cAMP, respectively. VIP demonstrated weak VPAC2 agonist activity, inducing a maximal 14-fold increase in cAMP. VIP4-28 had no VPAC2 agonist activity but demonstrated potent VPAC2 antagonist activity. VIP4-28 inhibited VPAC2-mediated increases in cAMP in Molt-4b cells up to 95%, but had no antagonistic effect on VPAC1. Lymphoblasts did not hydrolyze VIP4-28 to a form with VPAC1 antagonist activity. VIP4-28 thus is a lymphocyte-generated VIP fragment with potent agonist activity for VPAC1 and potent antagonist activity for VPAC2.

Cyanidin 3-O-β-D-glucoside isolated from skin of black Glycine max and other anthocyanins isolated from skin of red grape induce apoptosis in human lymphoid leukemia Molt 4B cells

Cyanidin 3-O-beta-D-glucoside (CG) was purified from black bean seed coat and other anthocyanins were prepared from red grape skin. These anthocyanins were identified by Mass, and 1H- and 13C-NMR. The effects of four anthocyanins on cell viability in human leukemia Molt 4B cells were investigated. The anthocyanins displayed strong growth inhibitory effects against human leukemia Molt 4B cells. Morphological changes showing apoptotic bodies were observed in the Molt 4B cells treated with these anthocyanins. The fragmentations by these anthocyanins of DNA to oligonucleosomal-sized fragments, that is a characteristic of apoptosis, were observed to be concentration-dependent. N-acetyl-L-cysteine, an antioxidant, suppressed the fragmentation of DNA by these anthocyanins. These findings suggest that growth inhibition of Molt 4B cells by these anthocyanins result from the induction of apoptosis by these compounds and that active oxygen is involved in the induction of apoptosis in the Molt 4B cells.

Diol- and triol-types of phytol induce apoptosis in lymphoid leukemia Molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to diol- and triol-types of phytol which were synthesized and identified by Mass, and 1H- and 13C-NMR, led to both growth inhibition and induction of programmed cell death (apoptosis). Morphological changes showing apoptotic bodies were observed in the Molt 4B cells treated with diol- and triol-types of phytol. The fragmentations of DNA by the diol- and triol-types of phytol to oligonucleosomal-sized fragments, that is a characteristic of apoptosis, were observed to be both concentration- and time-dependent. These findings suggest that growth inhibition of Molt 4B cells by the diol- and triol-types of phytol results from the induction of apoptosis in the leukemic cells.

Specific induction of apoptosis by 1,8-cineole in two human leukemia cell lines, but not a in human stomach cancer cell line

We have investigated the effects of 1,8-cineole [the main component of essential oil prepared from bay-leaves Laurus nobilis L.)] on DNA of human leukemia cell lines, Molt 4B, HL-60 and stomach cancer KATO III cells. Specific induction of apoptosis by 1,8-cineole was observed in human leukemia Molt 4B and HL-60 cells, but not in human stomach cancer KATO III cells. Morphological changes showing apoptotic bodies were observed in the human leukemia HL-60 cells treated with 1,8-cineole. The fragmentations of DNA by cineole to oligonucleosomal-sized fragments that is a characteristic of apoptosis were concentration- and time-dependent in Molt 4B and HL-60 cells, but not in KATO III cells. The present study shows that the suppression growth by 1,8-cineole in the leukemia cell lines results from the induction of apoptosis by this compound.

Selective induction of apoptosis by ar-turmerone isolated from turmeric (Curcuma longa L) in two human leukemia cell lines, but not in human stomach cancer cell line

We have investigated the effects of ar-turmerone isolated from turmeric (Curcuma longa L) on DNA of human leukemia cell lines, Molt 4B, HL-60 and stomach cancer KATO III cells. It was found that selective induction of apoptosis by ar-turmerone was observed in human leukemia Molt 4B and HL-60 cells, but not in human stomach cancer KATO III cells. Morphological changes showing apoptotic bodies were observed in the human HL-60 and Molt 4B cells treated with ar-turmerone. The fragmentation of DNA by ar-turmerone to oligonucleosomal-sized fragments that is a characteristic of apoptosis was observed to be concentration- and time-dependent in Molt 4B and HL-60 cells, but not in KATO III cells. The data of the present study show that the suppression by ar-turmerone of growth of these leukemia cell lines results from the induction of apoptosis by this compound.

Sesaminol from sesame seed induces apoptosis in human lymphoid leukemia Molt 4B cells

The exposure of human lymphoid leukemia Molt 4B cells to sesaminol, a component of sesame oil led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with sesaminol. The fragmentation of DNA by sesaminol to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition of Molt 4B cells by sesaminol results from the induction of apoptosis in the cells.

Sesamolin from sesame seed inhibits proliferation by inducing apoptosis in human lymphoid leukemia Molt 4B cells

The exposure of human lymphoid leukemia Molt 4B cells to sesamolin, a component of sesame seed led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with sesamolin. The fragmentation of DNA by sesamolin to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition of Molt 4B cells by sesamolin results from the induction of apoptosis in the cells.

Sodium 1-monolinolenin isolated from Italian ryegrass (Lolium multiflorum Lam) induces apoptosis in human lymphoid leukemia Molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to sodium 1-monolinolenin (SML) which was isolated from the leaves of Italian ryegrass (Lolium multiflorum Lam) and identified by Mass, and (1)H- and (13)C-NMR, led to both growth inhibition and induction of programmed cell death (apoptosis). Morphological change showing apoptotic bodies was observed in the Molt 4B cells treated with SML. The fragmentation by SML of DNA to oligonucleosomal-sized fragments, that is a characteristic of apoptosis, was observed to be both concentration- and time-dependent. These findings suggest that growth inhibition by SML of Molt 4B cells results from the induction of apoptosis in the cells.

Enin Isolated from Skin of Red Grape(Vitris vinifera) Induces Apoptosis in Human Lymphoid Leukemia Molt 4B Cells.

We have investigated the effect of extract prepared from skin of red grape (Vitris vinifera) on the growth of human lymphoid leukemia Molt 4B cells. It was found that the extract obtained by extraction with butanol suppressed the growth of Molt 4B cells. Fraction IV obtained by separation with HPLC inhibited the growth of the Molt 4B cells by 86% at the concentration of 600 μM. We determined the structure of compound contained in Fraction IV and identified it as enin, a kind of anthocyanin, by 1H- and 13C-NMR. The enin-treated cells clearly exhibited morphological changes, indicating the blebbing and chromatin condensation which are characteristic of apoptosis. Characteristic oligonucleosomal-sized DNA fragments were observed in the enin-treated cells. These findings suggest that the suppression by enin of growth of Molt 4B cells results from the induction of apoptosis by this compound.

Pheophorbide a, a moiety of chlorophyll a, induces apoptosis in human lymphoid leukemia molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to pheophorbide a (PPB a), a moiety of chlorophyll a, led both to growth inhibition and induction of programmed cell death (apoptosis). The growth inhibition by PPB a was much stronger than that by chlorophyll a. Morphological change showing apoptotic bodies was observed in the Molt 4B cells treated with PPB a. The fragmentation by PPB a of DNA to oligonucleosomal-sized fragments, that are characteristics of apoptosis, was observed to be concentration- and time-dependent. These findings suggest that growth inhibition by PPB a of Molt 4B cells results from the induction of apoptosis and that PPB a, moiety of chlorophyll a, is essential for exertion of antitumor and apoptosis-inducing activity in these cells.

Sesamin and episesamin induce apoptosis in human lymphoid leukemia Molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to sesamin and episesamin which were isolated from unroasted sesame seed oil and identified by MS, and 1H and 13C-NMR, led to both growth inhibition and the induction of programmed cell death (apoptosis). Morphological change showing apoptotic bodies was observed in the Molt 4B cells treated with sesamin and episesamin. The fragmentations by sesamin and episesamin of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis were observed to be concentration-dependent, respectively. Moreover, the amount of the DNA fragments in the sesamin-treated cells was increased from 2 days, while that in the episesamin-treated cells was elevated at 3 days after addition of the compounds. These findings suggest that growth inhibitions by sesamin and episesamin of Molt 4B cells result from the induction of apoptosis in the cells.

Sesamol Induces Apoptosis in Human Lymphoid Leukemia Molt 4B Cells.

The exposure of human lymphoid leukemia Molt 4B cells to sesamol, a component of sesame oil led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with sesamol. The fragmentation of DNA by sesamol to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. On the other hand, in the sesamol-treated normal human lymphocytes morphological change and DNA fragmentation were not observed at all. These findings suggest that growth inhibition of Molt 4B cells by sesamol results from the induction of apoptosis in the cells.

Phytol induces programmed cell death in human lymphoid leukemia Molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to phytol which was isolated from Lolium multiflorum Lam and identified by MS, and 1H- and 13C-NMR, led to both growth inhibition and the induction of programmed cell death (apoptosis). Morphological change showing apoptotic bodies was observed in the cells treated with phytol. The fragmentation by phytol of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition by phytol of Molt 4B cells results from the induction of apoptosis in the cells.

Effect of Eucommiol Isolated from Aucuba(Aucuba japonica Thunb) on the Growth of Human Lymphoid Leukemia Cells.

We have investigated the effect of crude extract prepared from leaves of Japanese aucuba (Aucuba japonica Thunb) on the growth of human lymphoid leukemia Molt 4B cells. The extract suppressed the growth of these cells by 60% at the concentration of 200 μg/ml. Fraction III obtained by separation with HW40S gel chromatography inhibited the growth of the cells by 95% at the same concentration. We analyzed the structure of the compound contained in Fraction III, and identified it as eucommiol by NMR and MS. The antiproliferative effect of eucommiol on the growth of Molt 4B cells was examined, and it was found to moderately suppress the proliferation of Molt 4B cells by 41% at the concentration of 200 μg/ml.

Honokiol induces apoptosis in human lymphoid leukemia Molt 4B cells.

The exposure of human lymphoid leukemia Molt 4B cells to honokiol led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with honokiol. The fragmentation by honokiol of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition by honokiol of Molt 4B cells results from the induction of apoptosis in the cells.

Effect of Oleanolic and Ursolic Acids Isolated from Loquat (Eriobotrya) on the Growth of Human Lymphoid Leukemia Cells.

We have identified two triterpene carboxylic acids of oleanolic acid and ursolic acid isolated from the leaves of loquat (Eriobotrya) by NMR spectroscopy. We have investigated the effects of oleanolic acid and ursolic acid on the growth of lymphoid leukemia Molt 4B cells and obtained results in which both compounds inhibited the proliferation of Molt 4B cells. In an in vitro experiment, ursolic acid inhibited ornithine decarboxylase (ODC) activity by 25.9% and 39.7% at the concentrations of 1 mM and 2 mM, respectively, while oleanolic acid inhibited ODC activity to a lesser extent than did ursolic acid. Both compounds slightly suppressed S-adenosylmethionine decarboxylase (AdoMetDC) activity. Polyamine contents were decreased in these compound-treated cells. These findings demonstrate that growth suppression of human Iymphoid Molt 4B cells by ursolic acid and oleanolic acid may result from the depletion of polyamines caused by the inhibition of ODC and AdoMetDC activities.

Inhibitory effects of persimmon (Diospyros kaki) extract and related polyphenol compounds on growth of human lymphoid leukemia cells.

We have investigated the effects of persimmon (Diospyros kaki) extract (PS) and related polyphenol compounds such as catechin (C), epicatechin (EC), epicatechingallate (ECG), epigallocatechin (EGC), and epigallocatechingallate (EGCG) on the growth of human lymphoid leukemia Molt 4B cells. We found that PS, ECG, EGC, and EGCG strongly inhibited the growth of the cells in a dose-dependent manner, while C and EC inhibited the growth of the cells only moderately. Ornithine decarboxylase (ODC), a rate-limiting enzyme of polyamine biosynthesis, was inhibited by 10-20% by these polyphenol compounds. The morphology of the Molt 4B cells indicated severe damage 3 days after treatment with PS, ECG, EGC, and EGCG. Irregular shape of the cells and DNA fragmentation were observed in PS, ECG, EGC, or EGCG-treated cells. These results suggest that PS, ECG, EGC, and EGCG induce apoptosis (programmed cell death) of Molt 4B cells.

Polyamine oxidase activity and growth in human cancer cells

lnterconversion of the polyamines, spermine + spermidine + putrescine, requires the N 1 -spermidine/spermine acetyltransferase (N1-SAT) and polyamine oxidase (PA01 enzymes. This process is used, in conjunction with biosynthesis and transport, to maintain the polyamine balance of the cell. N1-SAT is the rate-limiting step in the pathway and provides N1 -acetylpolyamine derivatives, the prefered substrate for PA0 [ll. PA0 contains a tightly bound FAD prosthetic group and acts on the secondary amine group of spermidine and spermine yielding spermidine and putrescine, respectively, along with 3-acetamidopropanal and hydrogen peroxide 121. Polyamine concentrations in cancerous tissue are high in comparison to normal tissue of the same type. High polyamine concentrations are also associated with rapidly dividing cells, suggesting alterations in either polyamine transport and/or polyamine metabolism are strongly associated with cell growth 131. As well as being substrates for PA0 acetylpolyamines have been suggested to be involved in excretion of polyamines 141. In this study we examined PA0 activity in a number of human tumour cell types and its relationship to cell growth. T47-D (breast carcinoma), G402 (renal leiomyoblastoma), and H T l l 5 (colonic carcinoma) cells were grown as monolayer cultures at a seeding density of 2 .5~104 celllcm2 (T47-D and G402) and 2.8~104 cell/cm2 (HT115). MOLT4-B (T leukaemia) cells were grown in suspension at 3x1 05 cells/ml. Culture medium was DMEM supplemented with 10% HS (HT1151, 10% FCS (G4021, 5% FCS & 1 % insulin (T47-D), or in RPMl with 10% FCS (MOLT4-B). Medium was changed every 48h where required. Cells were harvested at the appropriate growth condition for each cell type and the cells homogenised in Tris-HCI pH7.4 + 0.1 % Triton-X-l OO. PA0 activity was determined by a modification of the method of Table 1. PA0 activity in human tumour cells

Inhibitory effects of novel festuclavine derivatives on nucleoside uptake and incorporation into DNA and RNA in human lymphoid leukemia Molt 4B cells

Effects of festuclavine derivatives on nucleoside uptake by human lymphoid leukemia Molt 4B cells and on incorporation into TCA-insoluble materials in the cells were examined. The uptake and incorporation of uridine or thymidine were suppressed by festuclavine (EN01), 13-bromo-1-cyclopropylmethyl-festuclavine (EN02), 1-(4-chlorobenzenesulfonyl)festuclavine (EN03) and 1-cyclopentyl festuclavine (EN04) at 10–50 μM. Among these compounds, EN02 was most effective and at 50 μM it completely suppressed cellular uptake of the nucleosides and their incorporation into TCA-insoluble materials inhibiting the cellular proliferation. EN03 and EN04 moderately inhibited the transport and incorporation of the nucleosides in dose-dependent manners, while the mother compound EN01 had the least inhibitory effect. These findings indicated that alkylation at the indole nitrogen in combination with bromination at C-13 of the festuclavine molecule strengthened its inhibitory action on nucleoside uptake to a remarkable extent. The inhibition curves of nucleoside incorporation into TCA-insoluble materials showed quite similar dose-dependence to those of the inhibition curves for cellular nucleoside transport. These results suggest that the inhibitions of DNA and RNA syntheses by the festuclavine derivatives are due to the depressed transport of nucleosides into the leukemia cells.

Inhibition by polyamines of methylthiopropylamine-induced ornithine decarboxylase in human lymphoid leukemia Molt 4B cells

Methylthiopropylamine (MTPA), an inhibitor of spermidine synthase, markedly induced ornithine decarboxylase (ODC) activity (about 30-fold of the basal level) in human lymphoid leukemia Molt 4B cells. This induction was blocked by the addition of spermidine, spermine or putrescine simultaneously with MTPA. Inhibition by spermidine or spermine of the MTPA-induced ODC activity was larger than that by putrescine. The increase of ODC activity by MTPA led to the large increase of cellular putrescine content. This increase of putrescine content was abolished drastically by the simultaneous addition of spermidine or spermine. The increase of ODC activity was almost completely blocked by the addition of cycloheximide or actinomycin D. This finding suggested that the increase of ODC activity was not due to activation of ODC preformed in Molt 4B cells. The ODC induction by MTPA was dose-dependently blocked by adding the calcium channel blockers (verapamil and nifedipine) or protein kinase C inhibitors (1-(5-isoquinolinesulfonyl)-2-methylpiperazine and palmitoyl carnithine). These results suggested that calcium and protein kinase C (PKC) were involved in MTPA-associated induction of ODC.