Biomedical subjects

M M Ip

Publications and source records attributed to M M Ip.

At least 55 records · Page 3Linked to original sources

PubMed1991

The truncated glucocorticoid receptor in the P1798 mouse lymphosarcoma is associated with resistance to glucocorticoid lysis but not to other glucocorticoid-induced functions.

Glucocorticoid receptors in lines of the P1798 mouse lymphosarcoma either sensitive or resistant to glucocorticoid-induced lysis have been characterized and their functional significance determined. The glucocorticoid receptor from the cortisol-sensitive tumor is an Mr approximately 98,000 protein with a Stokes radius of 7.4 nm in the oligomeric, non-DNA-binding state and 5.6 nm in the transformed, DNA-binding state. This receptor binds glucocorticoid and reacts with the BUGR-2 monoclonal antibody. In contrast, two abnormal receptor species were identified in the cortisol-resistant tumor. One is an Mr approximately 98,000 non-steroid-binding but immunologically reactive protein. The other is an Mr approximately 45,000 species which contains both steroid- and DNA-binding sites but exhibits little or no reactivity with BUGR-2, suggesting that its NH2 terminus is truncated in a region within or adjacent to the BUGR epitope. This species had Stokes radii of 5.8 and 3.5 nm in nontransformed and transformed states, respectively. In both tumor lines, glucocorticoids stimulated the activities of glutamine synthetase and 5'-nucleotidase and the synthesis of glucocortin. However, glucocorticoid-induced tumor regression occurred only in the cortisol-sensitive tumor. Additionally, the glucocorticoid inducibility of a specific protein in the sensitive, but not in the resistant, tumor was demonstrated, as well as the presence of a protein specific to the resistant line. Taken together, these results suggest that the truncated glucocorticoid receptor in the P1798 lymphosarcoma is functional, although possibly in a more restricted gene-specific manner, and that the lysis defect, while possibly resulting from a truncated receptor, may also result from the inability of glucocorticoids to induce a critical protein in the pathway of programmed cell death and/or from the presence of a protein which inhibits the lytic response.

5'-Nucleotidase↗

PubMed1991

Mammary organoids from immature virgin rats undergo ductal and alveolar morphogenesis when grown within a reconstituted basement membrane.

We have recently described a primary culture system which allows for extensive proliferation and functional differentiation of immature mammary epithelial cells. Herein, these findings are extended to demonstrate that a distinct pattern of ductal and alveolar morphogenesis can be induced within the mammary organoids isolated from virgin female rats and cultured within an Engelbreth-Holm-Swarm sarcoma-derived reconstituted basement membrane under defined serum-free conditions. The lobular and multilobular organoids that emerged resemble the alveoli of the mammary gland in gross form, multicellular architecture, and cytologic and functional differentiation, while the ductal organoids expressed characteristics typical of mammary gland ducts in vivo. The epithelial cells within the alveolar- and duct-like organoids displayed the capability of secreting two morphologically distinct milk products, casein and lipid, into the luminal compartment. The expression of histiotypic morphogenesis and mammary-specific functional differentiation by the cultured mammary organoids proceeded in the absence of a morphologically distinct basal lamina. We illustrate that development highly reminiscent of that which naturally occurs in the mammary gland in vivo can be induced and supported in vitro under defined serum-free conditions. In addition, the methodologies are available to simultaneously monitor mammary organoid morphogenesis, growth, and functional differentiation. This system should serve as a unique model in which the regulation of branching morphogenesis, development, gene expression, and transformation can be examined.

Animals↗

PubMed1991

New site-directed polyclonal antibody maps N-terminus of occluded region of the non-transformed glucocorticoid receptor oligomer to within BUGR epitope.

Using a 17-mer synthetic peptide for immunization, a polyclonal antibody (WS933) directed against amino acid residues 395-411 of the mouse glucocorticoid receptor (GCR) has been raised and used to probe the significance of this region in forming the receptor oligomer and to localize the truncation site of the mutant GCR of the P1798 lymphosarcoma. This region of the receptor, which encompasses the BUGR epitope, is amino-terminal of and immediately adjacent to the DNA-binding domain. The polyclonal antibody WS933 reacted with both native and denatured forms of the wild-type mouse GCR as judged by its ability to shift the transformed receptor peak on Sephacryl S300 columns, to immunoadsorb the receptor to protein A Sepharose, and by immunoblot analysis where it identified the 98 kDa receptor protein in the cortisol-sensitive line of the P1798 mouse lymphosarcoma. WS933 also reacted with rat and rabbit GCR, but not human GCR. These characteristics were shared by the BUGR-2 monoclonal antibody. Unexpectedly, there were two highly significant differences between WS933 and BUGR-2. The first was the ability of WS933 to bind to the mutant 45 kDa GCR of the cortisol-resistant P1798 lymphosarcoma as judged by its capability of shifting the receptor peak on Sephacryl S300 columns. BUGR-2, in contrast, was unable to shift this mutant receptor peak. Secondly, WS933 was unable to react with the non-DNA-binding form of the wild-type (or mutant) GCR, whereas BUGR-2 could react with the non-DNA-binding form of the wild-type GCR. The first observation suggests that the truncation site of the mutant receptor may lie within a portion of the BUGR domain. Additionally, the second observation implies that at least part of the region lying within amino acid residues 395-411 of the mouse GCR is occluded in the receptor oligomer and that this site only becomes available upon transformation of the GCR to the DNA-binding form. This data provides the first mapping of the amino-terminus of the occluded region of the non-transformed receptor, and suggests that WS933 will be a useful probe for characterizing mutant as well as wild type glucocorticoid receptors.

Amino Acid Sequence↗

PubMed1990

Primary culture of normal rat mammary epithelial cells within a basement membrane matrix. I. Regulation of proliferation by hormones and growth factors.

A serum-free primary culture system has been developed which allows for three-dimensional growth and differentiation of normal rat mammary epithelial cells (RMECs) within an extracellular matrix preparation. RMECs were isolated from mammary glands of immature 50- to 60-d-old rats and the organoids embedded within a reconstituted basement membrane matrix prepared from the Engelbreth-Holm-Swarm sarcoma. Cells grown in a serum-free media consisting of phenol red-free Dulbecco's modified Eagle's medium-F12 culture medium containing 10 micrograms/ml insulin, 1 microgram/ml prolactin, 1 microgram/ml progesterone, 1 microgram/ml hydrocortisone, 10 ng/ml epidermal growth factor (EGF), 1 mg/ml fatty-acid-free bovine serum albumin (BSA), 5 micrograms/ml transferrin, and 5 microM ascorbic acid proliferated extensively (15- to 20-fold increase in cell number as quantitated using the MTT dye assay) over a 2- to 3-wk culture period and remained viable for months in culture. Several types of colonies were observed including the alveolarlike budding cluster which predominates at later times in culture, units with no or various degrees of ductal-like projections, stellate colonies, and two- and three-dimensional web units. Optimal proliferation required insulin, prolactin, progesterone, EGF, and bovine serum albumin. Hydrocortisone was not required for proliferation, but the colonies developing in its absence were morphologically altered, with a high frequency of colonies that formed an extensively branched network with many fine projections. Cell proliferation was also dependent on substratum, with significantly less growth and development occurring in RMECs grown within a type I collagen gel matrix compared to RMECs grown within the reconstituted basement membrane. In conjunction with other studies demonstrating extensive differentiation as well as proliferation, it is concluded that this model should prove to be an important tool to study the hormonal regulation of the growth and development of rat mammary cells.

Animals↗

PubMed1990

Primary culture of normal rat mammary epithelial cells within a basement membrane matrix. II. Functional differentiation under serum-free conditions.

A serum-free primary culture system is described which allows normal rat mammary epithelial cells (RMECs) embedded within a reconstituted basement membrane to undergo extensive growth and functional differentiation as detected by synthesis and secretion of the milk products casein and lipid. RMECs isolated from mammary glands of immature virgin rats were seeded within an extracellular matrix preparation derived from the Engelbreth-Holm-Swarm sarcoma and cultured in a serum-free medium consisting of Dulbecco's modified Eagle's medium-F12 containing insulin, prolactin, progesterone, hydrocortisone, epidermal growth factor, bovine serum albumin, transferrin, and ascorbic acid. Casein synthesis and secretion were documented at the electron microscopic level as well as by an enzyme-linked immunosorbent assay (ELISA) assay using a polyclonal antibody against total rat caseins. Numerous secretory vesicles with casein micelles were noted near the apical surface of the RMECs, and secreted casein was observed in the lumen. These ultrastructural data were confirmed by the ELISA assay which showed that microgram amounts of casein per well were synthesized by the RMECs and that the amount of casein increased with time in culture. Using immunoblot analysis it was demonstrated that the full complement of casein proteins was synthesized. In addition to casein protein, beta-casein mRNA levels were shown to increase with time. Synthesized lipid was detected at both the light and electron microscopic levels. Phase contrast photomicrographs demonstrated extensive intracellular lipid accumulation within the ductal and lobuloalveolarlike colonies, and at the electron micrograph level, lipid droplets were predominantly localized near the apical surface of the RMECs. The lipid nature of these droplets was verified by oil red O staining. Results from this study demonstrate that RMECs from immature virgin rats proliferate extensively and rapidly develop the capacity to synthesize and secrete casein and lipid when grown within a reconstituted basement membrane under defined serum-free conditions. This unique system should thus serve as an excellent model in which the regulation of mammary development and gene expression can be investigated.

Animals↗

PubMed1989

Short-term primary culture of rat prostate tumor epithelial cells on reconstituted basement membrane as a useful in vitro model to assess drug action on prostatic cancer: efficacy of the thiazolidinedione derivative CGP 19984.

A model system has been developed that permits short-term culture of rat R3327 prostate adenocarcinoma epithelial cells on a reconstituted basement membrane. Growth of prostate tumor cells under these conditions resulted in an enriched epithelial cell population that exhibited an eightfold increase in cell number in 10 days. This model system was used to test the efficacy of the thiazolidinedione derivative CGP 19984, a drug that inhibits luteinizing hormone secretion in vivo. At concentrations ranging between 1 and 25 micrograms/ml, CGP 19984 inhibited growth of the prostate tumor epithelial cells in a dose-dependent manner. The results thus demonstrate a direct effect of CGP 19984, which complements its indirect antitumor action in vivo, and suggest that this drug might be an effective agent for treatment of prostatic cancer. Moreover, growth of prostate tumor epithelial cells on a reconstituted basement membrane provides a useful system for in vitro testing of drugs for prostate cancer.

Adenocarcinoma↗

PubMed1989

Isolation and characterization of cortisol-sensitive and -resistant P1798 mouse lymphosarcoma cell lines.

Three cell lines have been isolated and characterized from the P1798 mouse lymphosarcoma. One line, derived from a glucocorticoid-resistant tumor, was glucocorticoid-resistant in vitro. The other two cell lines, derived from glucocorticoid-sensitive and -resistant parental tumors, respectively, were shown to be glucocorticoid-sensitive in vitro. The glucocorticoid receptor from all three cell lines bound glucocorticoid with similar affinity and capacity. However, based on Sephacryl S-300 gel filtration, the glucocorticoid receptor from the resistant cell line was smaller than that of the two sensitive cell lines. Moreover, the glucocorticoid receptor from the resistant cell line accumulated to a greater extent in the nucleus. This resistant cell line thus resembles the nti variant of the S49 lymphoma cell line. All three cell lines were tumorigenic and metastatic when reimplanted into mice, contained the normal mouse diploid complement of 40 chromosomes and exhibited the same responsiveness to cortisol in vivo as they did in vitro. It is concluded that the ready passage of these cell lines in vitro or in vivo and the presence of the small receptor in the resistant line should make them excellent model systems for the study of glucocorticoid resistance.

Animals↗

PubMed1989

A comparison of the effects of the prostaglandin synthesis inhibitors indomethacin and carprofen on 7,12-dimethylbenz[a]anthracene-induced mammary tumorigenesis in rats fed different amounts of essential fatty acid.

The effects of the cyclooxygenase inhibitors indomethacin and carprofen on the enhancement of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis by dietary linoleate have been compared in female Sprague-Dawley rats. Indomethacin and carprofen, 0.004% and 0.02% (w/w) in the diet, respectively, were fed to rats receiving 20% fat diets containing 0.5, 4 or 12% linoleate starting 7 days after administration of 5 mg DMBA i.g. Indomethacin was shown to have a marked inhibitory effect on mammary tumorigenesis in rats fed the 4 and 12% linoleate diets, but did not alter tumorigenesis in rats fed the 0.5% linoleate diet. In contrast, carprofen was not inhibitory in any of these dietary groups, or in a separate experiment in which a 5% fat--3% linoleate diet was fed. The effect of each drug on prostaglandin E2 (PGE2) levels in normal mammary glands enriched in epithelial cells after a 3-week pretreatment with 17 beta-estradiol and progesterone was also investigated. Carprofen was shown to reduce PGE2 levels to a similar or greater extent than indomethacin at each level of linoleate in the diet. These data demonstrate that a reduction in PGE2 synthesis in the mammary epithelium does not correlate with inhibition of mammary tumorigenesis, and that other factors, including possible alterations in other products of the arachidonic acid cascade, are responsible for this inhibitory effect.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1989

Exercise training and the differential prolactin response in male and female rats.

Adult male and female Sprague-Dawley rats were trained on a horizontal treadmill for 0, 1, 3, 5, or 7 days/wk for 10 wk. Speed and duration were progressively increased over 5 wk to a maximum of 20 m/min for 1 h. Between weeks 9 and 10 of training, animals were placed on the nonmoving treadmill, and blood (500 microliters) was sampled via chronic venous cannulas 30 min before, 0, 10, 20, 30, 45, and 60 min during exercise, and 15, 30, 60, 90, and 120 min after exercise. In another study, resting animals in the various groups were injected with thyrotropin-releasing hormone (TRH; 2 micrograms/kg for males and 0.4 microgram/kg for females) to determine pituitary prolactin responsiveness. In males, exercise induced a significant increase in plasma prolactin levels, with the greatest increase observed in the least trained and the smallest increase in the most highly trained animals. Female rats displayed the opposite trend with the greatest increase in prolactin secretion observed in the highest trained and the smallest increase observed in the least trained animals. TRH induced similar increases in plasma prolactin in all male groups, whereas TRH-induced prolactin release was greatest in the highest trained and smallest in the least trained females. The reduced prolactin response in highly trained males may reflect their acclimation to repetitive exercise stress, whereas the enhanced response in the highly trained female rats appears to result from increased pituitary sensitivity to prolactin-releasing factors.

Animals↗

PubMed1988

Combined therapy with 5-azacytidine and hydrocortisone in glucocorticoid-sensitive and -resistant mouse P1798 lymphosarcoma.

Experiments were undertaken to test the hypothesis that 5-azacytidine (AZA) treatment might make the glucocorticoid-resistant line of the mouse P1798 lymphosarcoma sensitive to cortisol. A modest (30%), though significant, effect in making cells cortisol sensitive was observed. Moreover, AZA enhanced tumor growth inhibition by cortisol in the corresponding sensitive line. The AZA alone was cytostatic, but it had no effect on DNA synthesis. The drug did not alter cytoplasmic glucocorticoid receptor levels or affinity, but nuclear levels were increased in the sensitive tumor. Results suggest that glucocorticoid resistance may be partially reversible and that the effect of AZA may be at the gene-expression level.

Animals↗

PubMed1988

Effects of the thiazolidinedione derivative CGP 19984 on growth and endocrine function of the MtT-W10 transplantable mammosomatotropic pituitary tumor in female rats.

The thiazolidinedione derivative CGP 19984 has previously been shown to suppress the growth of hormone-dependent mammary and prostatic tumors, primarily by reducing gonadotropin and subsequently gonadal steroid secretion. The present study examines the effects of CGP 19984 on the growth and hormone secretion of the autonomous, but estrogen-responsive, MtT-W10 mammosomatotropic transplantable rat pituitary tumor. Intact tumor-bearing Wistar/Furth female rats were administered vehicle or 25, 100, or 250 mg/kg CGP 19984 p.o., 5 x week for 4 weeks. CGP 19984 was found to significantly reduce MtT-W10 tumor growth and weight and reduce prolactin and growth hormone (GH) secretion in a dose-responsive manner. A similar study in ovariectomized rats also showed that CGP 19984 treatment suppressed MtT-W10 pituitary tumor growth, weight and hormone secretion in a dose-responsive manner, suggesting a direct inhibitory action of this drug on the tumor. In a third study, bromocryptine (CB-154; 5 mg/kg) and CGP 19984 (50 mg/kg) were both found to be effective in suppressing growth of the MtT-W10 tumor in intact female rats. However, rats treated with CGP 19984 alone had reduced serum and tumor GH and prolactin concentrations, while rats treated with CB-154 alone had reduced serum and tumor prolactin, but no change in GH concentrations. These results suggest that CGP 19984 effectively inhibits growth and hormone secretion of the autonomous MtT-W10 pituitary tumor by apparently suppressing both somatotropic and lactotropic cell populations within the tumor. Furthermore, these findings indicate that CGP 19984 may be an effective alternative to CB-154 in the clinical treatment of prolactin-producing adenomas, as well as other types of pituitary adenomas.

Animals↗

PubMed1987

Characterization of estrogen-binding proteins in variants of a rat mammary tumor.

Estrogen-binding proteins have been characterized in variants of the MTW-9B rat mammary tumor in an attempt to determine the functional significance of the low affinity cytosolic estrogen binder. In vivo selection of tumor variants was carried out by transplant of the tumor into intact and castrated male and female Wistar-Furth rats for four or five successive transplant generations. Tumors that developed in each of the four lines were taken for retransplant into intact and castrated male and female rats and all recipient tumor groups were compared for high and low affinity estrogen-binding proteins using isoelectric focusing analysis. No alterations in the isoelectric focusing profiles were observed in tumors that developed after repeated passage in ovariectomized female or castrated male rats when compared to the profile of the estrogen-binding proteins of the parent tumor carried routinely in intact female rats. However, a tumor variant containing only the low affinity, more basic estrogen-binding protein resulted from repeated passage of the MTW-9B mammary tumor in male rats. The high affinity estrogen receptor was absent in this variant and could not be induced by retransplant of the tumor into intact female hosts. Growth of the parent tumor and each of the variants was shown to be ovarian independent, suggesting that the presence of the low affinity estrogen-binding protein is not predictive of estrogen responsiveness. This suggestion is further supported by the observation that estrogen-stimulated progesterone receptor synthesis could be demonstrated in the parent tumor which demonstrated both binders, but not in the tumor variant developed in male rats which contained only the low affinity estrogen-binding protein. Progesterone receptor synthesis in this latter tumor appeared to be constitutive. Studies are continuing in an attempt to identify a role for the cytosolic low affinity estrogen-binding protein.

Animals↗

PubMed1987

Role of prolactin in modulating the effects of tamoxifen on growth of the Dunning R3327 rat prostate adenocarcinoma.

Tamoxifen (TAM) has previously been shown to inhibit growth of the Dunning R3327 rat prostate adenocarcinoma and to elevate serum prolactin levels. The purpose of this study was to determine the role of prolactin in modulating the effects of tamoxifen on growth of the R3327 prostatic adenocarcinoma. Intact and castrated Copenhagen-Fischer male rats bearing the Dunning R3327 rat prostatic tumor were divided into groups and injected sc five times per week for 16 weeks as follows: vehicle; TAM (0.5 mg/kg); haloperidol (HALO; 0.5 mg/kg); bromocriptine (CB-154; 5 mg/kg); TAM plus HALO; or TAM plus CB-154. In both intact and castrated rats, agents that either raised (HALO) or lowered (CB-154) serum prolactin had little effect on prostatic tumor growth when administered singly. In intact rats, average tumor diameter in vehicle-treated controls increased 421% 16 weeks after the start of the experiment, and treatment with TAM or TAM plus HALO reduced this tumor growth by approximately one-half. Interestingly, CB-154 administered in combination with TAM completely blocked TAM inhibition of tumor growth in intact rats. In contrast to these results in intact rats, average tumor diameter increased 129% in TAM- and 118% in TAM plus HALO-treated castrated rats and was significantly greater than the characteristic retardation of tumor growth (49% increase) that occurred in the vehicle-treated castrate controls. In addition, combined treatment of TAM plus CB-154 in castrate rats resulted in an even greater increase (188%) in average tumor diameter. The inhibitory effect of TAM on R3327 prostatic tumor growth in intact rats appears to be an indirect effect resulting from its ability to reduce serum testosterone levels. In contrast, the stimulatory effect of TAM in castrate rats appears to result directly from an estrogen-like action, which can directly enhance prostatic tumor growth in the presence of low levels of circulating androgens; this stimulatory effect of TAM is more pronounced when prolactin levels are suppressed by CB-154. Clearly, castration alone is more effective than TAM therapy alone or in combination with castration in the retardation of the growth of the androgen-dependent R3327 prostatic tumor in rats.

Adenocarcinoma↗

PubMed1987

Characterization and hormonal regulation of estrogen binding proteins in the MTW-9B transplantable rat mammary tumor.

A two-component estrogen (E2)-binding system has been characterized in the E2-independent MTW-9B rat mammary tumor by Scatchard analysis, sucrose gradient analysis, and isoelectric focusing. One cytosol receptor protein (type I) conforms to the classical estrogen receptor with a high affinity (Kd = 0.45 nM) for E2 and limited binding capacity [maximum binding (Bmax) = 53 fmol/mg protein]. The second component (type II) demonstrates a high number of sites (Bmax = 164 fmol/mg protein) and low E2-binding affinity (Kd = 22.3 nM). The type I and type II E2-binding components were shown to sediment on sucrose gradients at 9.6S and 4.5S, respectively, and to focus at isoelectric points of 6.6 and 8.0, respectively. The addition of 0.4 M KCl to the homogenization buffer converted the high affinity receptor species to a form that cosedimented and cofocused with the low affinity E2-binding protein. When tumors were grown in intact male rats, the ratio of the type II to the type I protein, as assessed by sucrose gradient analysis, increased 4.5-fold relative to that in tumors from intact females. Concomitantly, the Kd values of the type I and type II proteins were increased 9- and 2-fold, respectively, and the Bmax of the type I protein was decreased. No changes in the ratios of the E2-binding proteins were observed in tumors grown in ovariectomized female or castrated male rats; however, the Kd for both proteins was increased in tumors from the latter group. Relative to that in intact females, tumor growth was retarded in intact male rats, but was unaffected by ovariectomy or castration. These studies demonstrate that the presence of the low affinity E2-binding protein does not necessarily predict E2 responsiveness. While the role of the type II cytosolic protein has not yet been established, it is possible that it could act as a reservoir for E2, which is required to activate specific biochemical functions such as progesterone receptor synthesis. Alternatively, it could be a nonactivated, perhaps norphosphorylated, form of the E2 receptor which binds E2 with only a very low affinity.

Animals↗

PubMed1987

Characterization of natural cytotoxic effector cells isolated from rat liver.

Nonparenchymal liver cells (NPC) from normal untreated female Wistar/Furth rats were tested for natural cytotoxicity in a 4-hour 51Cr release assay against the murine lymphoma YAC-1, the murine mastocytoma P815, and the syngeneic rat mammary carcinoma TMT-081 tumor cell lines. NPC exerted strong cytotoxicity against all three target cells. In contrast, fresh spleen cells displayed cytotoxicity only against YAC-1, although after culture for 24 h at 37 degrees C cytotoxicity was displayed against all three target cells. Fresh spleen cells contained 2-15% large granular lymphocytes (LGL) as assessed by Giemsa staining whereas NPC contained 10-23% LGL and 10-25% Kupffer cells. Centrifugal elutriation produced fractions that were increased in one or the other of the cell types. More cytotoxic activity was observed in the fraction containing more LGL. The cytolytic activity of fresh spleen cells could be eliminated by either in vivo or in vitro treatment with anti-asialo-GM1 antiserum. On the other hand, the cytolytic activity of NPC was resistant to in vivo treatment, but was partially sensitive to in vitro treatment. Furthermore, the activity of cultured spleen cells was also partially sensitive to in vitro treatment. NPC and cultured spleen cells also were more resistant to suppression by prostaglandin E2 and nordihydroguaiaretic acid than fresh spleen cells. We conclude that LGL is mainly responsible for natural cytotoxicity of NPC and that some effector cells in NPC may be highly activated.

Animals↗

PubMed1986

Effect of dietary polyunsaturated fat and 7,12-dimethylbenz(a)-anthracene on rat splenic natural killer cells and prostaglandin E synthesis.

Dietary polyunsaturated fat has been shown to stimulate mammary tumorigenesis induced in rats by 7,12-dimethylbenz(a)anthracene (DMBA). Studies were undertaken to investigate the effect of polyunsaturated fat and DMBA on splenic natural killer (NK) activity and prostaglandin E (PGE) synthesis. In a first experiment, splenic NK activity at 33, 55, 75, and 110 days of age was measured in Sprague-Dawley rats fed 0.5% low fat (LF), 5% normal fat (NF), or 20% high fat (HF) corn oil diets from 23 days of age. At 55 days of age, half of the rats from the 75 and 110 day age groups were given 5 mg DMBA. Ten days after the initiation of the diets splenic NK activity against YAC-1 lymphoma was decreased from 50% cytotoxicity in rats fed NF diet to 21% cytotoxicity in rats fed HF diet, but was not affected by LF feeding. No difference in NK activity was observed among the groups at the later time periods. DMBA had no effect on NK activity at 20 or 55 days after its administration. In a second experiment, where DMBA (15 mg/rat) was given to half of the rats at 50 days of age and NF or HF diets were started 3 days later, NK activity was 35% in rats fed NF diet and 21% in rats fed HF diet, 5 days after the diets were started. No difference in NK activity in rats fed either diet was observed at later time periods. DMBA decreased both NK activity and spleen cellularity transiently. In both experiments, PGE synthesis by spleen cells cultured for 18 h was not affected by dietary fat intake, but was slightly increased 3 days after DMBA administration. Results from these experiments suggest that the stimulation of DMBA-induced mammary tumorigenesis by polyunsaturated fat and by DMBA itself may possibly be mediated by a transient decrease in splenic NK cell activity.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1986

Regulation of rat natural killing. II. Inhibition of cytolysis and activation by inhibitors of lipoxygenase: possible role of leukotrienes.

Rat splenic natural killer (NK) cell activity against 51Cr-labeled YAC-1 or TMT-081 tumor cells can be augmented by culturing at 37 degrees C for 18 hr. Inhibitors of the lipoxygenase pathway of arachidonic acid metabolism, NDGA, alpha-phenanthroline, quercetin, ETYA, BW755C, esculetin, and timegadine, inhibited this NK activation and also inhibited NK cytotoxicity when added directly to the NK assay. However, there was a partial loss of sensitivity of activated NK cells to suppression by NDGA, BW755C, and esculetin. Indomethacin failed to reverse the inhibition of NK activation caused by NDGA. However, LTB4 and LTC4 (0.01 microgram/ml) were able to reverse the inhibitory effect of NDGA on NK activation. Furthermore, spleen cells cultured for 18 hr synthesized detectable amount of LTC4 in their supernatants. NDGA inhibited the LTC4 synthesis in a dose-dependent manner. These data therefore suggest that leukotrienes are responsible for NK activation, and lipoxygenase activity is essential for NK cytolytic activity.

5,8,11,14-Eicosatetraynoic Acid↗

PubMed1986

Regulation of human natural killing. IV. Role of lipoxygenase in regulation of natural killing activity.

In this study we demonstrated that natural killer (NK) cell lysis by human peripheral blood nonadherent (NA) cells against K562 target cells was rapidly inhibited by four agents that inhibit the lipoxygenase pathway of arachidonic acid metabolism, nordihydroguaiaretic acid (NDGA), U-60257, alpha-phenanthroline, and esculetin. However, human NK cells activated by interferons (IFN) or poly I:C were partially resistant to suppression by NDGA and U-60257. Pretreatment of the NA cells with the four lipoxygenase inhibitors at 37 degrees C for 18 h led to suppression of NK activity. The inhibition of NK activity by NDGA was not reversed by aspirin at a concentration that inhibits PGE2 synthesis. Thus, suppression of NK activity by NDGA was not mediated by the effects on PGE2 synthesis. However, the inhibition of endogenous NK activity by NDGA, U-60257, alpha-phenanthroline, or esculetin was partially reversed by IFN or poly I:C. These results suggest that products of lipoxygenation are required for maintenance of human NK activity.

Arachidonate 12-Lipoxygenase↗