Vascular endothelial growth factor immunoneutralization in combination with cisplatin reduces EAC tumor growth

In the present study, we evaluated the effects of a neutralizing anti-Vascular Endothelial Growth Factor (VEGF) polyclonal antibody on murine EAC tumor growth both in vitro and in vivo. Furthermore, we investigated if in the presence of effective VEGF blockade, a conventional chemotherapeutic drug Cisplatin could be effective, and if so would there be an additive effect of the combination regimen. An in vitro cell proliferation assay using MTT kit showed that VEGF antibody alone inhibited proliferation of EAC cells significantly in all the three time intervals (p<0.05). But cisplatin treatment in combination with VEGF antibody resulted in highly significant inhibition (p<0.001) of cell proliferation. Apoptosis assay by FACS analysis showed that VEGF antibody-cisplatin combination treatment induced apoptosis in cultured EAC cells. Intraperitoneal administration of VEGF antibody (100 mug/dose) and cisplatin (0.5 mg/kg/dose) combination was observed to be more effective in reducing tumor burden and increasing life span when compared to VEGF antibody or cisplatin treatment alone in EAC solid tumor bearing mice. In EAC ascites tumor model, all the three types of treatment inhibited tumor burden and increased life span, but the inhibition was less compared to EAC solid tumor bearing mice. VEGF antibody singly and in combination with cisplatin reduced neoangiogenesis and vascular hyperpermeability. However, it is clear from the results that the combination treatment had no additive effect in reducing vascular hyperpermeability. Serum VEGF was not reduced significantly after treatment in EAC ascites tumor bearing mice, whereas in EAC solid tumor bearing mice it was reduced significantly after treatment. The results clearly show that though alone cisplatin showed antitumor efficacy but it had no significant inhibitory effect on neoangiogenesis and vascular hyperpermeability. Thus the present study suggests that anti-VEGF agent can be combined with traditional treatment modalities to ensure more effectiveness.     

Upregulation by glucocorticoids of responses to eosinopoietic cytokines in bone-marrow from normal and allergic mice

Since the production of eosinopoietic cytokines (GM-CSF, IL-3, IL-5) is inhibited by glucocorticoids, while responsiveness to these cytokines is enhanced in bone-marrow of allergic mice, we studied the ability of glucocorticoids to modulate murine bone-marrow eosinopoiesis. Progenitor (semi-solid) and/or precursor (liquid) cultures were established from bone-marrow of: (a) normal mice; (b) ovalbumin-sensitized and challenged mice or (c) dexamethasone (1-5 mg kg(-1)) injected mice. Cultures were established with GM-CSF (2 ng ml(-1)) or IL-5 (1 ng ml(-1)), respectively, alone or associated with dexamethasone, hydrocortisone or corticosterone. Total myeloid colony numbers, frequency and size of eosinophil colonies, and numbers of eosinophil-peroxidase-positive cells were determined at day 7. In BALB/c mice, dexamethasone (10(-7) M) increased GM-CSF-stimulated myeloid colony formation (P = 0.01), as well as the frequency (P=0.01) and size (P<0.01) of eosinophil colonies. Dexamethasone (10(-7) M) alone had no effect. Dexamethasone (10(-7)-10(-10) M) increased (P<0.002) eosinophil precursor responses to IL-5. Potentiation by dexamethasone was still detectable: (a) on low density, immature, nonadherent BALB/c bone-marrow cells, (b) on bone-marrow from other strains, and (c) on cells from allergic mice. Hydrocortisone and corticosterone had similar effects. Dexamethasone administered in vivo, 24 h before bone-marrow harvest, increased subsequent progenitor responses to GM-CSF (P = 0.001) and precursor responses to IL-5 (P<0.001). These effects were blocked by RU 486 (20 mg kg(-1), orally, 2 h before dexamethasone, or added in vitro at 10 microM, P<0.001). Glucocorticoids, acting in vivo or in vitro, through glucocorticoid receptors, enhance bone-marrow eosinopoiesis in na?ve and allergic mice.     

Trichosanthes dioica root extract induces tumor proliferation and attenuation of antioxidant system in albino mice bearing Ehrlich ascites carcinoma

Trichosanthes dioica Roxb. (Cucurbitaceae), called pointed gourd in English, is a dioecious climber grown widely in the Indian subcontinent. The present study assessed the influence of treatment of hydroalcoholic extract of Trichosanthes dioica root (TDA) on Ehrlich ascites carcinoma (EAC) in Swiss albino mice with effects on antioxidant systems. Twenty-four hours after intraperitoneal inoculation of tumor (EAC) cells in mice, TDA was administered at 25 and 50 mg/kg for 8 consecutive days. On the 9(th) day, half of the mice were sacrificed for estimation of tumor proliferation, hematological, and hepatic antioxidative parameters. The rest were kept for assessment of survival parameters. TDA exhibited dose dependent and significant increase in tumor weight, tumor volume, packed cell volume and viable cells and reduced non-viable cells and life span of EAC bearing animals. Hematological parameters were significantly worsened in TDA-treated mice. TDA treatment significantly aggravated the hepatic antioxidative parameters. The present study demonstrated that T. dioica root possessed tumor promoting activity in EAC bearing albino mice, plausibly mediated by attenuation of endogenous antioxidant systems.     

Dexamethasone exacerbates cisplatin‐induced muscle atrophy

Dexamethasone for antiemetic therapy is typically administered with anticancer drugs such as cisplatin. We previously reported that cisplatin upregulates the muscle‐specific E3 ubiquitin ligase genes, namely muscle ring‐finger protein 1 (MuRF1) and atrophy gene‐1 (atrogin‐1), and promotes muscle atrophy in mice. It is well known that dexamethasone causes upregulation of MuRF1 and Atrogin‐1 expression in skeletal muscles. Although it is speculated that a combination of dexamethasone and cisplatin worsens muscle atrophy, there are no reports based on research. We thereby investigated the effects of cisplatin and dexamethasone, alone or in combination, on the expression of MuRF1 and Atrogin‐1 in murine skeletal muscles and C2C12 myotubes. Mice were intraperitoneally injected with cisplatin or the vehicle control once daily for 4 days. Dexamethasone or the vehicle control was subcutaneously administered 30 minutes prior to the administration of cisplatin. Dexamethasone enhanced MuRF1 and Atrogin‐1 gene expression upregulated by cisplatin in murine quadriceps muscles and C2C12 myotubes. Cisplatin‐caused upregulation of myostatin and downregulation of IGF‐1 gene expression were also enhanced by co‐administration of dexamethasone in murine quadriceps muscles and C2C12 myotubes. This study shows that the combination treatment of cisplatin and dexamethasone exacerbated muscle atrophy in mice. Therefore, this treatment regimen might exacerbate muscle atrophy in cancer patients.     

Nerve Growth Factor from Cobra Venom Inhibits the Growth of Ehrlich Tumor in Mice

The effects of nerve growth factor (NGF) from cobra venom (cvNGF) on growth of Ehrlich ascites carcinoma (EAC) cells inoculated subcutaneously in mice have been studied. The carcinoma growth slows down, but does not stop, during a course of cvNGF injections and restores after the course has been discontinued. The maximal anti-tumor effect has been observed at a dose of 8 nmoles cvNGF/kg body weight. cvNGF does not impact on lifespan of mice with grafted EAC cells. K252a, a tyrosine kinase inhibitor, attenuates the anti-tumor effect of cvNGF indicating the involvement of TrkA receptors in the process. cvNGF has induced also increase in body weight of the experimental animals. In overall, cvNGF shows the anti-tumor and weight-increasing effects which are opposite to those described for mammalian NGF (mNGF). However in experiments on breast cancer cell line MCF-7 cvNGF showed the same proliferative effects as mNGF and had no cytotoxic action on tumor cells in vitro. These data suggest that cvNGF slows down EAC growth via an indirect mechanism in which TrkA receptors are involved.     

地塞米松脂质体的制备及其对乳腺癌作用的体内外研究

目的制备地塞米松脂质体,探讨地塞米松对乳腺癌4T1细胞的生长抑制作用及对荷瘤鼠的抗肿瘤药效。方法采用薄膜分散–超声法,以粒径和多分散指数(PDI)为指标进行单因素实验考察了大豆磷脂(SPC)与甲氧基聚乙二醇磷脂(DSPE-mPEG2000)的质量比、SPC与地塞米松的质量比、超声时间对地塞米松脂质体粒径的影响从而筛选得到最佳处方和最佳工艺条件。采用MTT法比较地塞米松注射液和地塞米松脂质体对4T1细胞的作用。建立4T1 BAL B/c荷瘤小鼠模型,研究地塞米松脂质体对4T1荷瘤小鼠的体内抗肿瘤作用。结果当SPC与DSPE-mPEG2000质量比为5∶1、SPC与地塞米松质量比为50∶3、超声时间为20 min时制备得到的脂质体粒径最小,粒径分布最窄,室温放置15 d稳定,于生理介质中稳定。MTT测定结果显示地塞米松注射液和脂质体对4T1细胞生长抑制作用均较弱,但在4T1荷瘤鼠的体内实验中,在5mg/kg的给药剂量下,地塞米松脂质体的抑瘤率却高达78.9%,显著高于地塞米松注射液(33.4%,P<0.05)和8mg/kg紫杉醇注射液(55%,P<0.05)。结论制备的地塞米松脂质体放置于生理介质中均能稳定存在,能口服能静脉给药。地塞米松脂质体对4T1荷瘤小鼠肿瘤生长有较强的抑制作用,但体外对4T1细胞抑制抑制作用并不强,推测地塞米松脂质体是通过调节肿瘤微环境来抑制肿瘤生长。     

Dexamethasone reduces morphine-induced Straub reaction in mice

This study examined the effect of dexamethasone on morphine-induced straub reaction in mice. When morphine was administered in doses of 7.5, 15 and 30 mg kg(-1) intraperitoneally, a dose-dependent straub reaction was produced. Dexamethasone per-se (0.1-10 mg kg(-1) i.p.) did not modify the tail of control mice. Pre-treatment with dexamethasone 120 min before morphine injection caused a dose-dependent reduction of straub reaction. Cycloheximide (15 mg kg(-1) i.p.) administered 2 h before morphine did not change morphine-induced straub reaction, but was able to prevent the effects of dexamethasone on morphine-induced straub reaction. The glucocorticoid receptor antagonist RU-38486 (15 mg kg(-1) i.p.) did not affect morphine-induced straub reaction, whereas it was able to block the effects of dexamethasone on morphine-induced straub reaction. Results of this study indicate that dexamethasone reduced morphine-mediated straub reaction in mice, indicating a further important functional interaction between dexamethasone and the opioid system. The ability of cycloheximide and RU-38486 to block dexamethasone's effects indicates that the steroid's interference with morphine-mediated straub reaction involves a protein-synthesis-dependent mechanism via glucocorticoid receptors.     

Dexamethasone Reduced Clonidine-induced Hypoactivity in Mice

Reduced clonidine anti-nociception in mice given low doses of dexamethasone has encouraged us to investigate the effects of dexamethasone pretreatment on locomotor hypoactivity, another example of clonidine-induced behaviour in mice. Dexamethasone administered intraperitoneally (0.1, 1.0, 10 mg kg^?1) 30 min before clonidine reduced clonidine-induced locomotor hypoactivity in the activity cage to an extent which was dose-dependent. Dexamethasone administered centrally (10 ng/mouse) 30 min before clonidine was also able to reduce clonidine-induced locomotor hypoactivity. Cycloheximide administered at a dose of 10 mg kg^?1 2 h before clonidine did not change the effects of clonidine but was able to prevent the effects of dexamethasone on clonidine-induced hypoactivity. The glucocorticoid receptor antagonist RU38486 administered centrally at the dose of 1 ng/mouse did not change the effects of clonidine, whereas it was able to block the effects of dexamethasone on clonidine-induced locomotor hypoactivity. These results suggest that the effects of dexamethasone on clonidine-induced locomotor hypoactivity depend on the stimulating effects that dexamethasone exerts on the protein synthesis via the glucocorticoid receptor in the brain.     

Clonidine-induced antinociception and locomotor hypoactivity are reduced by dexamethasone in mice

The effects of dexamethasone pretreatment on clonidine-induced antinociception and locomotor hypoactivity were investigated in mice. In the hot-plate and the tail-flick tests, dexamethasone administered intraperitoneally at a dose of 1 mg kg(-1), 30 or 60 min before clonidine, reduced clonidine antinociception in both tests and reduced clonidine-induced locomotor hypoactivity in the activity cage. When administered 15 min before clonidine, dexamethasone had no effect on clonidine antinociception. A higher dexamethasone dose (10 mg kg(-1)) induced the same effects observed at a dose of 1 mg kg(-1) in the hot-plate and the tail-flick tests, but the former dose had a stronger effect on locomotor hypoactivity. Dexamethasone (10 ng/mouse) administered intracerebroventricularly 30 min before clonidine was also able to reduce both clonidine-induced antinociception and locomotor hypoactivity. The protein synthesis inhibitor, cycloheximide, administered intraperitoneally at the dose of 10 mg kg(-1), 2 h before clonidine, was able to prevent dexamethasone effects on clonidine-induced antinociception. The glucocorticoid receptor antagonist RU-38486, administered intracerebroventricularly at the dose of 1 ng/mouse, was also able to block dexamethasone effects on clonidine-induced antinociception and locomotor hypoactivity, whereas both cycloheximide and RU-38486 per se did not influence pain sensitivity or locomotor activity. These results suggest that the dexamethasone effects on clonidine-induced antinociception and locomotor hypoactivity depend on the stimulating effects that dexamethasone exert, on the protein synthesis via the glucocorticoid receptor in the brain.     

女贞子多糖抗肿瘤作用研究

目的探讨女贞子多糖(LLP)的抗肿瘤作用及机制。方法观察LLP对小鼠肉瘤(S180)、小鼠肝癌(H22)的抑制作用;采用MTT法观察LLP对人肝癌细胞(SMMC-7721)增殖的影响;ConA刺激T-淋巴细胞法研究LLP的免疫刺激作用。结果LLP对S180、H22实体瘤均有抑制作用(P<0.05);对SMMC-7721肿瘤细胞无直接杀伤作用,可提高Co-nA对T淋巴细胞刺激的转换率,提高由淋巴细胞YAC-1所致NK细胞的活性。结论LLP具有抗实体肿瘤的作用,LLP抗实体瘤的作用与其提高机体免疫,改善机体免疫能力而抑制肿瘤细胞生长有关。     

Antitumor and antioxidant activity of Polyalthia longifolia stem bark ethanol extract

In the present study, the ethanol extract of stem bark of Polyalthia longifolia Benth. and Hook (Annonaceae) was screened for its in vitro and in vivo antitumor activity. In vitro cytotoxicity of P. longifolia extract was assessed in murine cancer cells and in human cancer cells by Trypan blue exclusion assay and MTT assay, respectively. P. longifolia extract showed concentration-dependent cytotoxicity in Ehrlich’s ascites carcinoma (EAC) and Dalton’s ascites lymphoma (DLA) cells with IC₅₀ values of 45.77 and 52.52?µg/mL, respectively. In the MTT assay, the IC₅₀ values of P. longifolia extract against HeLa and MCF-7 cells were 25.24 and 50.49 µg/mL, respectively. In vivo antitumor activity against Ehrlich’s ascites tumor and Dalton’s solid tumor models was assessed by administering 50 and 100?mg/kg of P. longifolia extract, i.p., for 7 consecutive days. P. longifolia extract, at a dose of 100?mg/kg, significantly enhanced mean survival time (MST) and marginally improved hematological parameters when compared to EAC control mice. And the same dose significantly reduced the tumor volume as compared to control DLA inoculated mice. Positive control, cisplatin (3.5?mg/kg, i.p., single dose), significantly enhanced MST and improved hematological parameters when compared to EAC and significantly reduced the tumor volume when compared to DLA control. In vitro antioxidant potential of P. longifolia extract was also determined owing to the role of reactive oxygen species in tumor initiation and progression. P. longifolia extract scavenged DPPH radicals, reduced ferric ions and inhibited lipid peroxidation with IC₅₀ values of 18.14, 155.41 and 73.33 µg/mL, respectively.     

乙烷硒啉与顺铂或氟尿嘧啶联合应用对胃癌BGC-823的治疗作用

目的观察乙烷硒啉与顺铂或氟尿嘧啶联合应用对胃癌BGC-823的体内、外抗肿瘤作用。方法MTT法测定5、10、20、40μmol·L~(-1)乙烷硒啉单药及固定其浓度为5μmol·L~(-1)与顺铂或氟尿嘧啶联合时对BGC-823的生长抑制作用。建立裸鼠移植瘤模型,采用乙烷硒啉、顺铂、氟尿嘧啶单药及乙烷硒啉与后两者联合的不同给药方案,观察抗肿瘤作用。结果乙烷硒啉对BGC-823有明显增殖抑制作用,24、48、72h的IC_(50)值分别是30.23、19.70和11.67μmol·L~(-1),联合后效果明显增强(P<0.05)。体内实验,联合组肿瘤增长率明显降低,与单药组有非常显著差异(P<0.01)。结论乙烷硒啉单药表现明显抗肿瘤作用,与顺铂或氟尿嘧啶联合后可以增效。     

Anti-tumor effect of gallic acid on LL-2 lung cancer cells transplanted in mice.

We previously reported that gallic acid (3,4,5-trihydroxybenzoic acid), a naturally occurring plant phenol, can induce apoptosis in four kinds of human lung cancer cell lines in vitro. The present study further investigated the in vivo anti-tumor effects of orally administered gallic acid. Gallic acid reduced cell viability of LL-2 mouse lung cancer cells in vitro dose dependently, with a 50% inhibitory concentration (IC50) value of around 200 microM. C57Black mice were transplanted with LL-2 cells, and administered gallic acid (1 mg/ml in drinking water, ad libitum) and/or cisplatin (4 mg/kg i.p. injection, once a week). The average weight of the transplanted tumors, obtained at 29 days after transplantation, in the mice of control, gallic acid-treated cisplatin-treated and cisplatin plus gallic acid-treated groups was 4.02, 3.65, 3.19 and 1.72 g, respectively. The average tumor weight of the mice treated with cisplatin combined with gallic acid was significantly smaller than that of the control group (p<0.05). The amount of apoptotic cells in the tumor tissues of mice treated with gallic acid and/or cisplatin was significantly higher than those of the control mice. Combination of gallic acid and cisplatin increased the tumor cell apoptosis compared with the treatment with cisplatin alone. The present findings suggest that the combination of gallic acid with an anti-cancer drug, including cisplatin, may be an effective protocol for lung cancer therapy.     

Adrenal corticosteroids as antiemetics during cancer chemotherapy

Adrenal corticosteroids were first reported in 1979 to have antiemetic effects during cancer chemotherapy. Since then considerable numbers of trials have been conducted to evaluate their activity alone and in combination with other agents. The majority of the research has centered on dexamethasone, although other corticosteroids have been studied. Dexamethasone as a single agent is superior to placebo and appears to be more effective than standard doses of prochlorperazine when administered with highly emetic agents. Dexamethasone is comparable to metoclopramide against moderately emetogenic agents and low-dose cisplatin, but less effective than metoclopramide against highly emetic agents or high-dose cisplatin. Dexamethasone improves the activity of prochlorperazine and metoclopramide and may reduce some of the side effects associated with the latter. Current trials continue to explore the role of corticosteroids alone and in combination with antiemetics.     

硫酸壳聚糖体内抗肿瘤作用的实验研究

目的：研究硫酸壳聚糖的体内抗肿瘤作用。方法：用高、低（200、100mg/kg）两个剂量的硫酸壳聚糖分别腹腔注射治疗肉瘤180（S180）小鼠和艾氏腹水癌（EAC）小鼠10d,然后测定其抑瘤率、重要器官的内脏指数和生命延长率,同时设生理盐水组（空白对照组）和氟尿嘧啶组（阳性对照组）进行比较。结果：硫酸壳聚糖高、低剂量组和氟尿嘧啶组的抑瘤率分别为38.67%、30.19%和43.27%,3组的生命延长率分别为65.38%、69.23%和54.93%。和生理盐水组、氟尿嘧啶组相比,硫酸壳聚糖高、低剂量组的S180小鼠的胸腺指数均有明显增加（P〈0.05）。结论：硫酸壳聚糖能有效抑制S180小鼠肿瘤的生长和延长EAC小鼠的生存时间,其作用机制可能与其提高机体的免疫力有关。     

The therapeutic efficacy of angiostatin against weakly- and highly-immunogenic 3LL tumors

BACKGROUND: Antiagiogenesis represents a promising approach to cancer therapy. We have previously demonstrated that the antitumor effects of endostatin, one of the most potent angiostatic agents, could be enhanced when combined with immunotherapy. Our current study evaluated whether anti-tumor immune response could also potentiate the therapuetic efficacy of another antiangiogenic drug, angiostatin. METHODS AND RESULTS: Using Matrigel assay, we showed that our preparation of recombinant angiostatin possessed potent anti-angiogenic activity in vivo. The antitumor effects of recombinant angiostatin were tested against weakly-immunogenic 3LL Lewis lung carcinoma versus its highly immunogenic variant 3LL-C75. We showed that angiostatin inhibited the growth of 3LL-C75 more potently than that of 3LL tumor, suggesting that the host's immune response potentiates the antitumor effects of angiostatin. This conclusion was further supported by the finding that the antitumor activity of angiostatin against 3LL-C75 tumor was lower in immunodeficient nude mice in comparison with immunocompetent mice. Immunization of C57BL/6 mice with 3LL-C75 cells stimulated the antitumor immunity and inhibited the growth of parental 3LL tumor. Angiostatin treatment of immunized mice further enhanced the antitumor effect of tumor vaccination. CONCLUSION: Antitumor immune response could complement the therapeutic efficacy of angiostatin.     

Augmented antitumor effects of combination therapy with VEGF antibody and cisplatin on murine B16F10 melanoma cells

Our previous studies with EAC tumor model demonstrated that a VEGF polyclonal antibody combined with cisplatin inhibited tumor growth. Here we report the antitumor effect of VEGF antibody plus cisplatin on a murine metastatic tumor model specially emphasizing its effect on different angiogenic parameters both in vitro and in vivo. Mouse B16F10 melanoma cells were cultured in vitro in DMEM media containing 10% FBS, nonessential amino acids and antibiotics in a 5% CO(2) incubator at 37 degrees C and the effect of VEGF antibody singly and in combination with cisplatin on this cell was assessed by MTT assay, matrigel invasion study and MMP-9 expression study in vitro. In vivo studies were performed by two tumor models viz B16F10 solid tumor model and B16LuF10 lung tumor model. The mice treated with VEGF antibody (PAb) alone, cisplatin alone and combination of VEGF antibody and cisplatin on alternative days from the next day of tumor transplantation. Antitumor as well as antiangiogenic efficacy was monitored by measuring tumor burden, survivability, MVD measurement, serum NO value measurement and bcl-2 expression study. It was observed that administration of combined therapy with VEGF antibody and cisplatin augmented antitumor activity in B16F10 melanoma models than the either agents alone. Thus our experiments show a successful VEGF antibody based combination therapy with cisplatin and suggests that the enhancement of antitumor activity could be explained by a concomitant effect on both endothelial and tumor cell compartment.     

Antiemetic effect of dexamethasone on cisplatin-induced early and delayed emesis in the pigeon

We investigated the ability of dexamethasone to attenuate cisplatin (4 mg/kg, i.v.)-induced early and delayed emesis. These appear within the first 8-h period (early phase) and between 8 and 48 h (delayed phase), respectively, after cisplatin administration in the pigeon. Dexamethasone (0.1 and 1 mg/kg, i.m.) reduced significantly the number of emetic responses to cisplatin by 56% and 82% (P<0.05), respectively, in the early phase, and by 41% and 66% (P<0.05), respectively, in the delayed phase. Dexamethasone (1 and 10 microg/kg, i.c.v.) reduced the number of emetic responses by 66% and 91% (P<0.05), respectively, in the early phase, and by 56% and 87% (P<0.05), respectively, in the delayed phase. Indomethacin (10 mg/kg, i.m.) did not suppress cisplatin-induced early and delayed emesis. Dexamethasone (1 mg/kg, i.m.) did not affect the content of platinum in the medulla oblongata after cisplatin administration. The above results suggest that dexamethasone has antiemetic effects on both the early and delayed emetic responses to cisplatin in pigeons, partially via its central site of action, and that the antiemetic mechanism of dexamethasone is related to factors other than its inhibition of prostanoid synthesis or its membrane stabilizing effect which reduces influx of cisplatin into the medulla oblongata.     

Glucocorticoid hormones prevent the induction of gamma-glutamyl transpeptidase by ethanol in a rat hepatoma cell line

The increase in serum gamma-glutamyl transpeptidase (GGT) is a well known marker of chronic alcoholism in man. We have previously shown that ethanol (180 mM) induces GGT activity 2-3-fold in the C2 rat hepatoma cell line. In this study, we have analyzed the interaction of ethanol with steroid hormones and drugs in this well defined cell culture system. Dexamethasone (100 nM), a synthetic glucocorticoid agonist, completely prevented the induction of GGT by ethanol, but had no effect when added alone. This inhibitory effect was also observed with other corticosteroids, but not with sex steroids; it was prevented by RU 486, a glucocorticoid antagonist. These observations suggest that dexamethasone acts through a high affinity glucocorticoid receptor. Conversely, ethanol did not interfere with the glucocorticoid induction of alanine aminotransferase in the same cell. We have analyzed the metabolism of ethanol in the C2 cells. These cells lack significant alcohol dehydrogenase activity as well as any cytochrome P-450 Alc immunoreactivity. Dexamethasone did not modify the disappearance of ethanol in the culture medium of those cells. We conclude that glucocorticoid hormones interact with ethanol at the cellular level, and that this interaction does not involve a modification of alcohol metabolism.     

Anti-tumor effect of 3-amino-N-substituted-pyrrolidine-2,5-dione-N-mustard hydrochloride.

The anti-tumor effect of 3-amino-N-substituted pyrrolidine-2,5-dione-N-mustard hydrochloride (PNM.HCl) against Ehrlich (ascites) carcinoma (EAC) was studied. A substantial increase in the survival of mice bearing EAC tumor was achieved following daily administration of PNM.HCl at subtoxic dosages. The therapeutic efficacy of PNM.HCl was maintained with changes in dosages and the schedules of administration. The effect of PNM.HCl when administered with conventional anti-cancer drugs at different time schedules against EAC was also studied. The results demonstrated an augmentation of anti-tumor activity in the case of certain anti-cancer drugs against EAC tumor, thereby suggesting a potential usefulness of PNM.HCl in multi-drug therapy.