Uptake Of Adriamycin Research Articles

Cytocidal Activity of a Synthetic Isoprenoid, N‐Solanesyl‐N, N′‐bis(3,4‐dimethoxy‐benzyl)ethylenediamine, and Its Potentiation of Antitumor Drugs against Multidrug‐resistant and Sensitive Cells in vitro

A synthetic isoprenoid, N‐solanesyl‐N, N′‐bis(3,4‐dimethoxybenzyl)ethylenediamine (SDB‐ethylene‐diamine), inhibited the colony formation of multidrug‐resistant mutant cell lines derived from Chinese hamster V79 (V79/ADM) and human hepatoma PLC/PRF/5 (PLC/COL) cells to a greater extent than that of the parental cells. When combined with other clinically useful antitumor agents, it potentiated the cytotoxic activity of almost all kinds of drugs tested including adriamycin (ADM), actinomycin D, vincristine, cytosine arabinoside, and 5‐fluorouracil (5‐FU), and the potentiation ratios were higher against V79/ADM cells than against V79/S cells. Among the antitumor agents tested, the activities of bleomycin‐group antibiotics were more strongly enhanced by SDB‐ethylenediamine and the potentiation was higher in the parental cells than in V79/ADM cells. SDB‐ethylenediamine enhanced the uptake of ADM and daunorubicin into V79/ADM and its parental cells, but it did not increase the uptake of 5‐FU or peplomycin, indicating that different mechanisms operate for potentiation in the cases of the latter drugs, i.e., not simply an increase of intracellular drug uptake. Two fragments of SDB‐ethylenediamine, solanesol (polyprenoid moiety) and the diamine component (verapamil‐like moiety), showed neither cytotoxic activity nor potentiator activity, even if they were mixed together, indicating that the steric conformation of intact SDB‐ethylenediamine molecule is important for these two activities.

Sensitivity to thermochemotherapy of AKR lymphoma and B16 melanoma variants of malignancy

Drug resistance, which so often accompanies tumor progression, has been shown to be related to changes in membrane properties which may result in decreased drug accumulation in the tumor cell. A correlation between sensitivity to thermochemotherapy and degree of malignancy was found in the AKR lymphoma system. Hyperthermia increased adriamycin (ADR) uptake and concomitantly its cytotoxicity to AKR lymphoma cells. Moreover, these effects were more pronounced on a variant of high malignancy (HM) than on a low malignancy (LM) one. Fluorescent microscopy, as well as cytofluorometry, indicated that lymphoma cells treated by ADR at 43 degrees C were more permeable to the cytotoxic agent than those exposed to the chemotherapeutic substance at 37 degrees C. Cytofluorometry indicated the presence of a minor cell subpopulation with low ADR uptake in the HM variant, not found in the LM one. Fluorocytometry also showed that the temperature-dependent increased ADR uptake was more marked in the HM than in the LM variant, explaining the differential effect of thermochemotherapy on the two lymphoma variants. However, correlation between degree of malignancy and sensitivity to thermochemotherapy is not a general feature. In contrast to the results obtained in the AKR lymphoma system, in the B16 melanoma the low malignancy variant, F1, was more markedly affected by the combined treatment than the F10 variant. The increased cytotoxic effect of ADR by supranormal temperatures in the F1 variant was shown to be due to an augmented drug uptake. The results suggest that drug resistance in late stages of tumor progression can be overcome by an agent acting on the cell membrane. However, the data also indicate the necessity of assaying cancer treatment modalities, including those designed to circumvent drug resistance, on various tumor system models.

長鎖下飽和脂肪族化合物による抗癌剤の耐性克服とその機構

We have attempted the overcoming of anticancer drug resistance in murine leukemia by unsaturated aliphatic compouns and investigated the drug up-take into cells using adriamycin(ADM)resistant mouse P 388 leukemia(P 388/ADM). When 150 μM unsaturated fatty acids such as oleic acid(OA, C18 : 1 Δ9), linoleic acid(LA, C18 : 2 Δ9-12)and α-linolenic acid(α-LNA, C18 : 3 Δ9-12-15)were added along with ADM to the P 388/ADM or ADM sensitive P 388 (P 388/S)culture in vitro, ADM resistance was greatly reduced, and especially α-LNA completely overcomed the resistance. In addi-tion, α-LNA showed a stronger its own killing activity to P 388/ADM than OA and LA. These fatty acids also potentiated the cytotoxicity of ADM against P 388/S. The uptake of ADM was increased by these fatty acids, however, large dif-ferences of enhancing rate at 3h were observed between P 388/S(2-7 fold)and P 388/ADM(16-30 fold). These findings suggest that unsaturated fatty acids can overcome ADM resistance in P 388 leuke-mia by enhancing the drug uptake and its own cell killing activity.

HPLC and Flow Cytometric Analyses of Uptake of Adriamycin and Menogaril by Monolayers and Multicell Spheroids

We have used both HPLC and flow cytometry to measure and compare the uptake of two anthracyclines, menogaril (MEN) and Adriamycin (ADR), in V79 Chinese hamster lung fibroblasts grown as monolayers and as 650 microns multicell spheroids. In order to compare intracellular drug accumulation in spheroid cells measured by the two methods, we converted mean channel fluorescence of the flow cytometer to drug uptake expressed as ng/10(6) cells by using a standard curve. The standard curve related the flow cytometric mean channel fluorescence, of monolayer cells exposed to either drug, to the intracellular drug accumulation determined by HPLC. This standard curve was then used to convert the mean channel fluorescence of cells from drug-exposed spheroids to ng/10(6) cells. Our results show that equal intracellular drug accumulation (determined by HPLC) in spheroids and monolayers does not result in equal cellular fluorescence emission (determined by flow cytometry) by these 2 cell populations. For example, monolayer cells with an intracellular MEN accumulation of 650 ng/10(6) cells, emit 40 units of fluorescence as measured by flow cytometry. However, spheroid cells with the same intracellular accumulation emit about 80 units of fluorescence. This results in the intracellular MEN uptake in spheroids measured by flow cytometry being as much as 2- to 3-fold higher than that measured by HPLC. Intracellular ADR accumulation measured by flow cytometry was also higher than that obtained by HPLC. In spite of the quantitative difference between the two methods, qualitatively both methods gave similar results. Thus, both techniques showed that at equal drug concentration in medium drug uptake in monolayers was much greater than in spheroids.(ABSTRACT TRUNCATED AT 250 WORDS)

ラット膀胱における Adriamycin の経膀胱的吸収と消失に関する蛍光顕微鏡的研究

The absorption and disappearance of adriamycin (ADM) through the bladder epithelium in the rat was investigated using a fluorescence microscope. In the absorption study in vitro, ADM permeated through the epithelium into the lamina propria or the inner layer of the muscle within 15 minutes after instillation, showing no further infiltration into the deeper part of the bladder wall. In addition, fluorescence histological evidence for the uptake of ADM by the endothelial cells of the blood vessels in the lamina propria suggested a systemic diffusion of ADM to the whole blood by blood circulation. These data coincided well with the absorption study in vivo. On the other hand, ADM once absorbed in the bladder wall disappeared from the lamina propria or the inner layer of the muscle and was confined only in the epithelium at 60 min after the excretion of ADM. The intensity of ADM fluorescence then decreased gradually with time elapsed. No ADM fluorescence was recognized in the bladder tissue 48 hr later. In the absorption study with the BBN induced rat bladder tumor, ADM permeated into all the cells of papillary tumor within 5 minutes of instillation. The fluorescence histological method used in the present study is harmless and may readily be applicable to the pharmacodynamic study of ADM in human bladder tumors. This method proves to be a promising approach to the development of an ideal instillation therapy effective on bladder tumor cells without side-effects.

Visualization of adriamycin uptake and internalization by receptor-mediated endocytosis in mast cells. an ultrastructural study using adriamycin-colloidal gold complexes

Visualization of adriamycin uptake and internalization by receptor-mediated endocytosis in mast cells. an ultrastructural study using adriamycin-colloidal gold complexes

Influence of interferon on adriamycin uptake of cultured tumor cells

Effect of human interferon beta (IFN beta) or interferon gamma (IFN gamma) on cellular uptake of adriamycin (ADM) was measured in 3 in vitro-cultured human tumor cell lines. Pre-incubation of tumor cells with 100 IU/ml of IFN beta or gamma for 6 hr resulted in an increased intracellular ADM concentration, but ADM efflux was not affected by the IFNs. The increase in cellular uptake of ADM was in agreement with both DNA and protein synthesis inhibition. 3H thymidine (3H-TdR) incorporation was remarkably inhibited by ADM with pre-treatment of 100 IU/ml of each IFN, and an additive effect of ADM and IFN on cell growth inhibition could be observed. Similar additive inhibition against tumor-cell proliferation in nude mice, which received combined treatment (twice/week) of IFN gamma (5,000 IU/mouse) and ADM (5-20 micrograms/mouse), was also observed. These results indicate that IFN increases cellular influx of ADM, and antiproliferative activity against tumor cells is enhanced by a combination of IFN and ADM.

Effect of phosphatidylserine on adriamycin uptake and histamine release in rat peritoneal mast cells

Effect of phosphatidylserine on adriamycin uptake and histamine release in rat peritoneal mast cells

Interaction of adriamycin with rat and mouse mast cells: Histamine release and cellular uptake

In the present study, we evaluated in mouse and rat mast cells if adriamycin uptake is related to histamine release. In addition, the uptake of the antineoplastic drug and histamine release were studied in other normal and neoplastic cell lines. The effect of sodium cromoglycate was also examined. Adriamycin induced histamine release from mixed or purified mast cells of the mouse and rat. This exocytotic response was quantitatively related to intracellular concentrations of adriamycin. Significant differences in adriamycin uptake were observed between mast cells and other normal and neoplastic cells. When peritoneal mast cells were treated with sodium cromoglycate, histamine release and adriamycin uptake were significantly reduced. In contrast, incubation of other cells with cromoglycate only slightly reduced the cellular concentration of the antineoplastic drug.

Effects of adriamycin and etoposide on the replication of adenovirus 5 in sensitive and resistant human tumour cells

Adenovirus is a potential probe for identifying and understanding drug sensitivity in primary, nonproliferating cultures of human normal and tumour cells but the scope and limitations of such an approach first need to be evaluated in established cell lines. For this purpose we have identified an ovarian tumour cell line (CI-80-13S) with natural resistance to adriamycin, etoposide and crosslinking agents compared with other human tumour lines. Resistance to adriamycin correlated poorly with resistance to etoposide in these cell lines ( r = 0.05). Adenovirus replication in drug-treated cells (viral capacity) was found to be differentially inhibited in sensitive cells when the drug was administered to cells simultaneously with infection (adriamycin) or 20 hr after infection (etoposide). Viral capacity could not be inhibited by more than 90% in sensitive cells. In contrast, no such plateau was exhibited in the dose-responses of cell survival or inhibition of cellular DNA synthesis, both of which distinguished sensitive from resistant cells. Adenovirus was not inactivated by preincubation with high doses of adriamycin or etoposide, thus confirming that no functionally-relevant damage is directly induced by these agents in DNA. Uptake of adriamycin and etoposide was similar in sensitive and resistant cells and both agents blocked cells in the G2 phase of the cell cycle. Protein-linked DNA was induced in sensitive cells. The results indicate that (a) these drugs have two dose-dependent effects in cells, one of which does not inhibit replication of adenovirus; and (b) inhibition of adenovirus replication could in principle be used to predict sensitivity to adriamycin and etoposide.

Ehrlich腹水癌細胞の薬剤耐性細胞における温熱によるAdriamycinの効果増強とその修飾

Ehrlich腹水癌細胞の薬剤耐性細胞における温熱によるAdriamycinの効果増強とその修飾

Modulation of adriamycin uptake by lonidamine in ehrlich ascites tumor cells

The effect of Lonidamine, 1-(2,4 dichlorobenzyl)-1-H-indazol-3-carboxylic acid, on the uptake of Adriamycin by Ehrlich ascites tumor cells has been investigated. The uptake of Adriamycin is greatly stimulated by Lonidamine and the increase depends on the energy sources of the cell. In the presence of glucose the intracellular drug content is remarkably lower than that in its absence. This difference lies in the mechanism by which Lonidamine enhances the uptake of Adriamycin. The Adriamycin efflux is via an active transport process and, in the presence of glucose, both aerobic glycolysis and oxidative phosphorylation contribute to ATP synthesis. Although Lonidamine inhibits both these pathways, there is still sufficient ATP to extrude a certain amount of Adriamycin. The elevated intracellular concentration of Adriamycin depends not only on the Lonidamine-inhibited outward transport but also on higher membrane permeability which allows a low concentration of Adriamycin (18 μM) to interfere also with the oxidative metabolism of Ehrlich ascites tumor cells.

Effect of sodium cromoglycate on adriamycin uptake in mast cells and other cell types

Effect of sodium cromoglycate on adriamycin uptake in mast cells and other cell types

Organ distribution of Adriamycin� after intravesical instillation with or without Tween 80 in the rat

The absorption of Adriamycin (ADM) into the systemic circulation and into different organs including the urinary bladder was investigated in the rat after intravesical instillation of ADM with or without a surface active detergent, Tween 80. The postinstillation plasma concentration of ADM increased significantly with increasing dose of the drug. Even though the leakage of ADM into the systemic circulation and into extravesical organs in general was slight a significant increase was observed after addition of Tween 80. The uptake of ADM into the urinary bladder wall was also significantly enhanced by Tween 80 thus the role of this agent in conjunction with intravesical chemotherapy should be further investigated.

Membrane transport changes in an adriamycin-resistant murine leukemia cell line and in its sensitive parental cell line.

Multidrug resistance in cancer chemotherapy occurs when cells develop resistance towards structurally and functionally unrelated drugs. It is speculated that alteration of some fundamental process(es) in the cells leads to the development of multidrug resistance. The sodium pump activity of murine leukemia cell lines P388/S (sensitive) and P388/ADR (resistant) was measured and found to be different in the two cell lines. The rate of sodium pumping, i.e., the ouabain-sensitive rubidium uptake, was consistently lower in the resistant cells compared to their parental controls. Uptake of adriamycin was lower in the resistant cells. Depolarizing the cells with potassium chloride or by inhibiting the pump with ouabain increased the adriamycin uptake in the sensitive cells but not in the resistant cells. Adriamycin did not have any acute effects on the sodium pump activity. It is concluded that the development of drug resistance in cell line P388 is associated with a decrease in sodium pump activity and a lack of depolarization-induced adriamycin uptake; these processes may be causally linked via alterations in cytosolic calcium concentration.

Improved Sulfatide-Containing Liposomes as Carriers of Adriamyein

Various molar ratios of the constituents of sulfatide-containing liposomes (egg phosphatidylcholine, cholesterol, and sulfatide) were examined with respect to liposomal efficiency to entrap adriamycin (ADM) and to the tissue distribution. Among the ratios of the constituents examined, 5:4:1 was the best in terms of entrapment efficiency for both multilamellar vesicles (MLV) and small unilamellar vesicles (SUV). Tissue distribution studies of liposome-entrapped ADM showed that the blood clearance and cardiac uptake of ADM were significantly reduced when the drug was entrapped in the above liposomes, especially in the MLV. Incorporation of ADM into ovarian tumors transplanted into nude mice was increased when the drug was entrapped in the SUV.

The relationship between membrane permeability to adriamycin and adriamycin cytotoxicity in CHO cells at elevated temperatures

The cytotoxicity of adriamycin at elevated temperatures was studied in CHO cells in vitro using a clonogenic assay. There is a significant increase in the cytotoxicity of adriamycin at temperatures which are not lethal to mammalian cells (38°C–42°C). The uptake of [ 14C]adriamycin was studied in similar conditions to determine whether the thermal enhancement of adriamycin cytotoxicity was due to an increase in membrane permeability. Although measurable increases in adriamycin uptake occur in the elevated but non-lethal temperature range, they are insufficient to explain the observed increase in cytotoxicity.

The effect of exposure to elevated temperatures on membrane permeability to Adriamycin in Chinese hamster ovary cells in vitro

The effect of environmental temperature on plasma membrane permeability to Adriamycin was studied in Chinese hamster ovary (CHO) cells in vitro using flow cytometry. Initial rates of uptake increased steadily between 23°C and 47°C and there was a greater than 2-fold increase in permeability between 37°C and 47°C. The increase in permeability with increasing temperature was greater than expected based on the model of passive drug influx. Adriamycin uptake was also measured at 37°C following previous exposure of the cells to elevated temperatures. Twenty-minute preexposures to temperatures above 41°C caused a significant decrease in membrane permeability, which fell to approximately 60% of control levels after exposure to 45°C. Longer-periods of pre-exposure to temperatures as low as 40°C were also shown to decrease membrane permeability to Adriamycin subsequently measured at 37°C. The state of decreased permeability to Adriamycin induced by hyperthermia persisted for at least 2 h. The thermally induced decrease in membrane permeability to Adriamycin is of potential importance to the design of optimal schedules for thermochemotherapy.

Modification of cellular efflux and cytotoxicity of adriamycin by biscoclaulin alkaloid in vitro

The intracellular uptake, retention and cytotoxicity of adriamycin (ADR) combined with a biscoclaulin alkaloid, cepharanthine, were investigated by flow cytometry in NIH 3T3 cells. Cepharantine suppressed the efflux of ADR in a similar fashion to verapamil. The intracellular uptake and retention of ADR were increased gradually by 0.1 to 1 μg/ml of cepharanthine and reached a plateau at > 1μg/ml. Cepharanthine, which had no toxic action on survival, increased intracellular ADR uptake by about 20% for 1 h incubation at 37 °C, and increased cellular ADR retention after incubation in an ADR-free medium for 4 h from 15% to 75%. The cytotoxicity of ADR was enhanced 5-fold in the cells pre- and co-incubated with cepharanthine. When cepharanthine was present in the medium before, during and for colony formation (10 days) after incubation with ADR, the cytotoxicity increased to about 300-fold. Furthermore, an increase in intracellular uptake of ADR was induced by an elevated temperature of 43 °C, and the efflux of ADR was inhibited by cepharanthine. A high level of intracellular ADR was maintained during the treatment. These results suggest a possible novel use of cepharanthine to improve the drug sensitivity of tumors resistant to ADR.

The effect of the non-ionic surfactant brij 30 on the cytotoxicity of adriamycin in monolayer, spheroid and clonogenic culture systems

The effects of a non-ionic polyoxyethylated lauryl ether surfactant (Brij 30) on monolayer uptake and spheroid penetration of adriamycin have been studied. Co-incubation of adriamycin with Brij 30 increases intracellular adriamycin levels by 2–3-fold. Although, in the concentrations used, Brij 30 alone is not cytotoxic, adriamycin and Brij 30 mixtures are significantly more cytotoxic (monlayer id 90 = 0.6 μg/ml; disaggregated spheroid id 30 = 1.9 μg/ml) and induce significantly longer spheroid growth delay than adriamycin alone (monolayer id 90 = 2.1 μg/ml; disaggregated spheroid id 50 = 3.3 μg/ml). Adriamycin is equally cytotoxic to mouse normal granulocytes and chronic myeloid leukaemic (M1 cell line) cells in agar clonogenic cultures. The addition of Brij 30 appears to enhance preferentially the activity of adriamycin against these tumour cells relative to the normal granulocytes.