ACTH 1-24 inhibits proliferation of adrenocortical tumors in vivo

Objectives: Although several lines of evidence suggest that the overall effects of the ACTH receptor, melanocortin 2 receptor (MC2-R), mediated signal transduction on adrenocortical growth and tumorigenesis are anti-proliferative, activation of MC2-R induces mitogens like jun, fos, and myc and activates the MAPK pathway. In vivo, potential effects of endogenous ACTH on adrenal tumori-genesis can not be separated from effects of other POMC derived peptides. Methods: Murine adrenocortical tumor cells that lack MC2-R expression (Y6pcDNA) and Y6 cells stablely transfected with MC2-R (Y6MC2-R) were generated. Presence of functional MC2-R was demonstrated by RT-PCR and Western blot using an antibody for phosphorylated CREB. As a syngenic tumor model, LaHeF1/J mice simultaneously received 107 Y6MC2-R and Y6pcDNA subcutaneously, giving rise to MC2-R positive and negative tumors within the same animal. Animals were treated for 3 weeks in groups of 12 according to the following schedule: group A, control animals receiving saline injection; group B, animals receiving 5.7 ng/injection of a slow release formula of ACTH 1-24 administered i.p. three times a week (aiming at a low physiologic dose); and group C, animals receiving 57 ng/injection of ACTH 1-24 (high physiological dose). Results: Twenty days of ACTH 1-24 treatment did not significantly affect corticosterone levels, endogenous ACTH levels or adrenal and thymus weight compared with saline injection. However, ACTH 1-24 treatment of group B and C mice significantly reduced tumor weight in MC2-R positive tumors in a dose dependent manner (P = 0.03), while no significant difference in tumor mass was observed in MC2-R negative tumors. PCNA and TUNEL staining, together with morphological characterization, demonstrated that these in vivo effects were due to reduced proliferation, while apoptosis and cellular hypertrophy within the tumor remained unchanged. Conclusion: MC2-R expression is associated with a less aggressive adrenal tumor phenotype and anti-proliferative effects can be amplified through stimulation with physiological doses of ACTH.

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Immunomodulatory activities of whey fractions in efferent prefemoral lymph of sheep

Journal of Dairy Research ◽

10.1017/s0022029900031757 ◽

1996 ◽

Vol 63 (2) ◽

pp. 257-267 ◽

Cited By ~ 12

Author(s):

Chun W. Wong ◽

Geoffrey O. Regester ◽

Geoffrey L. Francis ◽

Dennis L. Watson

Keyword(s):

Immune Responses ◽

Bovine Milk ◽

Inhibitory Effect ◽

Dependent Manner ◽

Milk Whey ◽

Significant Difference ◽

Lymphocyte Phenotypes ◽

Dose Dependent

SummaryStudies on the immunomodulatory activities of ruminant milk and colostral whey fractions were undertaken. By comparing with boiled colostral whey in a preliminary experiment, a putative heat-labile immunostimulatory factor for antibody responses was found to be present in ovine colostral whey. Studies were then undertaken in sheep in which the efferent prefemoral lymphatic ducts were cannulated bilaterally, and immune responses in the node were measured following subcutaneous injection in the flank fold of whey protein preparations of various purities. A significant sustained decline of efferent lymphocyte output was observed following injection with autologous crude milk whey or colostral whey preparations, but no changes were observed in interferon-gamma levels in lymph plasma. Two bovine milk whey fractions (lactoperoxidase and lactoferrin) of high purity were compared in bilaterally cannulated sheep. A transient decline over the first 6 h was seen in the efferent lymphocyte output and lymph flow rate after injection of both fractions. A significant difference was seen between the two fractions in interferongamma levels in lymph at 6 h after injection. However, no significant changes in the proportion of the various efferent lymphocyte phenotypes were seen following either treatment. Whereas both fractions showed a significant inhibitory effect in a dose-dependent manner on the proliferative response of T lymphocytes, but not B lymphocytes, to mitogenic stimulation in vitro, no similar changes were seen following in vivo stimulation with these two fractions.

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Changes in pituitary responsiveness to LH-releasing hormone after acute buserelin treatment: a time-course and dose–response study

Journal of Endocrinology ◽

10.1677/joe.0.1060027 ◽

1985 ◽

Vol 106 (1) ◽

pp. 27-30 ◽

Cited By ~ 1

Author(s):

J. D. Heather ◽

S. A. Whitehead

Keyword(s):

Time Course ◽

Dependent Manner ◽

Releasing Hormone ◽

Serum Lh ◽

Significant Difference ◽

Lh Release ◽

Lh Releasing Hormone ◽

In Vivo Effects ◽

Dose Response Study

ABSTRACT The acute in-vivo effects of a potent LH-releasing hormone (LHRH) agonist, buserelin, on LH secretion and pituitary responsiveness to LHRH have been investigated in oestrous rats. Doses of 50, 100 and 250 ng buserelin stimulated LH release in a dose-dependent manner, the peak serum LH concentrations being measured 1 h after the treatment. Thereafter LH levels fell rapidly between 1 and 6 h and by 18 h serum LH concentrations were similar in all groups of animals. Pituitary responsiveness to a challenge with 100 ng LHRH was potentiated by 50 or 100 ng buserelin injected 1 or 2 h before the LHRH challenge. In contrast, 250 ng buserelin completely abolished the LH response to LHRH when tested 1, 2 and 4 h after treatment, but by 6 h a small but attenuated response was observed. Four hours after treatment there was no significant difference in the responses when compared with the saline-treated controls. J. Endocr. (1985) 106, 27–30

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Effect of Single Dose In Vivo Propranolol Therapy on In Vitro Adhesion of Human SS RBC.

Blood ◽

10.1182/blood.v108.11.1234.1234 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1234-1234 ◽

Cited By ~ 1

Author(s):

Laura M. De Castro ◽

Jude C. Jonassaint ◽

Jennifer G. Johnson ◽

Milena Batchvarova ◽

Marilyn J. Telen

Keyword(s):

Safety Data ◽

Study Drug ◽

Flow Chamber ◽

In Vivo Studies ◽

Dependent Manner ◽

Number Of Patients ◽

Significant Difference ◽

Before And After

Abstract Sickle red blood cells (SS RBC) are abnormally adhesive to both endothelial cells (ECs) and components of the extracellular matrix (ECM). Epinephrine (epi) has been shown to elevate cAMP in SS RBC and increase adhesion of SS RBC to ECs in a protein kinase A-dependent manner. In vitro and in vivo studies performed in our lab have led to the hypothesis that adrenergic stimuli such as epi may initiate or exacerbate vaso-occlusion and thus contribute to the association of vaso-occlusive events with physiologic stress. We are conducting a prospective, dose-escalation pilot clinical study to investigate whether in vivo administration of one dose of propranolol either down-regulates baseline SS RBC adhesion in vitro or prevents its upregulation by epi. In addition, this study will provide additional safety data regarding the use of propranolol in normotensive patients with sickle cell disease (SCD). Figure Figure To date, we have completed the first two dose cohorts. 11 subjects (9 SS and 1 Sβ° thalassemia; 7 females, 3 males) have participated. No severe adverse events were noted. Cohorts 1 and 2 had mean pre-propranolol blood pressure (BP) of 116 (5.9 SD)/ 60.4 (3.98 SD) and 106.8 (4.68 SD)/ 58 (3.9 SD), respectively; this difference was not statistically significant. Minimal and asymptomatic changes in BP were noted in both cohorts after drug administration, with biphasic systolic and diastolic BP nadirs at 45 and 240 minutes. No clinically significant changes in heart rate were observed. Adhesion studies were performed using a graduated height flow chamber on the day of RBC collection. RBC adhesion to ECs was studied before and after epi stimulation and was measured at sheer stresses ranging from 1 to 3 dyne/cm2. Baseline adhesion measurements were validated by comparing percent (%) adhesion assayed at 2 different times within 7 days—at screening and before propranolol dose on the study drug day. We observed no significant difference in adhesion at the 2 different time points without propranolol. Comparison of % adhesion of epi-stimulated RBC to ECs before and 1 hour after propranolol showed that propranolol given in vivo significantly inhibited both non-stimulated and epi-stimulated SS RBC adhesion (p=0.04 and p=0.001, respectively). Lastly, comparison of SS RBC adhesion at both drug doses confirmed the drug-related inhibition of adhesion (p<0.004). We conclude that propranolol administered in vivo decreases SS RBC baseline adhesion to ECs and substantially abrogates epi-stimulated adhesion to ECs, as measured in vitro. Although we have thus far studied only a small number of patients and low propranolol doses, we expect to confirm these results with the 3rd cohort, in which a higher dose of propranolol will be used. If our findings continue to show that propranolol can decrease both SS RBC baseline and epi-stimulated adhesion to ECs, study of propranolol on a larger scale would be warranted in order to ascertain its safety and efficacy as an anti-adhesive therapy in SCD.

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Periplocin Extracted from Cortex Periplocae Induced Apoptosis of Gastric Cancer Cells via the ERK1/2-EGR1 Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000445555 ◽

2016 ◽

Vol 38 (5) ◽

pp. 1939-1951 ◽

Cited By ~ 20

Author(s):

Lei Li ◽

Lian-Mei Zhao ◽

Su-li Dai ◽

Wen-Xuan Cui ◽

Hui-Lai Lv ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Viability ◽

Cancer Cells ◽

Inhibitory Effect ◽

Tunel Staining ◽

Dependent Manner ◽

Gastric Cancer Cells ◽

Induced Apoptosis

Background/Aims: Periplocin is extracted from the traditional herbal medicine cortex periplocae, which has been reported to suppress the growth of cancer cells. However, little is known about its effect on gastric cancer cells. Methods: Gastric cancer cells were treated with periplocin, and cell viability was assessed using MTS assay. Flow cytometry and TUNEL staining were performed to evaluate apoptosis, and protein expression was examined by western blotting. Microarray analysis was used to screen for changes in related genes. Results: We found that periplocin had an inhibitory effect on gastric cancer cell viability in a dose-dependent manner. Periplocin inhibited cell viability via the ERK1/2-EGR1 pathway to induce apoptosis. Periplocin also inhibited the growth of tumor xenografts and induced apoptosis in vivo. Conclusion: Our results show that periplocin inhibits the proliferation of gastric cancer cells and induces apoptosis in vitro and in vivo, indicating its potential to be used as an antitumor drug.

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Zoledronic Acid Inhibits Osteoclastogenesis in Vitro and in a Mouse Model of Inflammatory Osteolysis

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348940311200907 ◽

2003 ◽

Vol 112 (9) ◽

pp. 780-786 ◽

Cited By ~ 16

Author(s):

Holger Sudhoff ◽

Brian T. Faddis ◽

Jae Y. Jung ◽

Henning Hildmann ◽

Jörg Ebmeyer ◽

...

Keyword(s):

Zoledronic Acid ◽

Bone Marrow Cells ◽

Mineral Apposition Rate ◽

Tunel Staining ◽

Tartrate Resistant Acid Phosphatase ◽

Mouse Bone Marrow ◽

Dependent Manner ◽

Dose Dependent

This study assessed effects of the bisphosphonate zoledronic acid (ZLNA) on osteoclastogenesis. To assess the effect of ZLNA on osteoclast formation in vitro, we cultured mouse bone marrow cells under conditions that promote osteoclastogenesis. Administered at concentrations from 10−6 to 10−9 mol/L, ZLNA led to a dose-dependent inhibition of osteoclastogenesis. Combined TUNEL staining and histochemical staining for tartrate-resistant acid phosphatase showed that ZLNA induced apoptosis in osteoclasts and monocytic precursor cells. To study the effects of ZLNA in vivo, we placed keratin particles onto the surface of the parietal bone of mice to induce localized inflammatory bone resorption. Three experimental groups received daily subcutaneous injections of ZLNA (1, 3, or 10 μg/kg body weight) from 4 days before surgery until 5 days after keratin implantation. The ZLNA significantly reduced osteoclast recruitment in a dose-dependent manner, but did not affect the degree of inflammation or the mineral apposition rate.

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Exosomal miR-1290 promotes angiogenesis in hepatocellular carcinoma via targeting SMEK1

10.21203/rs.2.13182/v1 ◽

2019 ◽

Author(s):

Qiong Wang ◽

Guanwen Wang ◽

Lianjie Niu ◽

Shaorong Zhao ◽

Jianjun Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Endothelial Cells ◽

Tumor Angiogenesis ◽

Molecular Mechanisms ◽

Target Genes ◽

Primary Liver Cancer ◽

Tumor Model ◽

Luciferase Reporter ◽

Dependent Manner

Abstract Abstract Background: Hepatocellular carcinoma (HCC), the most common primary liver cancer, rely on the formation of new blood vessel for growth and frequent intrahepatic and extrahepatic metastasis. Therefore, it is important to explore the underlying molecular mechanisms of tumor angiogenesis of HCC. Recently, microRNAs have been shown to modulate angiogenic processes by modulating the expression of critical angiogenic factors. However, the potential roles of tumor-derived exosomal microRNAs in regulating tumor angiogenesis remain to be elucidated. Methods: MiRNome sequencing was performed to uncover the miRNAs that are dysregulated in HCC patient serum-derived exosomes. Expression levels of miR-1290 in tissues and cells were determined by quantitative real-time PCR. The effect of mir-1290 on proliferation was evaluated by CCK-8 assay. The angiogenic ability of cells were determined by transwell, wound-healing, tube formation and matrigel plug assays. SMMC-7721 xenograft tumor model was established in NOD-SCID nude mice using miR-1290 and NC antagomirs to determin the angiogenic effect of mir-1290 in vivo. Target protein expression was determined by western blotting. Dual luciferase reporter assay was performed to confirm the action of miR-1290 on downstream target genes including SMEK1. Results are reported as means ± S.D. and differences were tested for significance using 2-sided Student’s t-test. Results: In this study, our miRNome sequencing demonstrated that miR-1290 was overexpressed in HCC patient serum-derived exosomes, and we found that delivery of miR-1290 into human endothelial cells enhanced their angiogenic ability. Our results further revealed that SMEK1 is a direct target of miR-1290 in endothelial cells. MiR-1290 exerted its pro-angiogenic function, at least in part, by inhibiting the VEGFR2 signaling pathway in a SMEK1-dependent manner. Conclusions: Collectively, our findings provide evidence that miR-1290 is overexpressed in HCC and promotes tumor angiogenesis via exosomal secretion, implicating its potential role as a therapeutic target for HCC.

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Epiregulin induces leptin secretion and energy expenditure in high-fat diet-fed mice

Journal of Endocrinology ◽

10.1530/joe-18-0289 ◽

2018 ◽

Vol 239 (3) ◽

pp. 377-388 ◽

Cited By ~ 1

Author(s):

Rumana Yasmeen ◽

Qiwen Shen ◽

Aejin Lee ◽

Jacob H Leung ◽

Devan Kowdley ◽

...

Keyword(s):

Energy Expenditure ◽

High Fat Diet ◽

Ex Vivo ◽

Mapk Pathway ◽

Treated Group ◽

Dependent Manner ◽

High Fat ◽

Fat Depots ◽

Expression Of Genes

Adipokine leptin regulates neuroendocrine circuits that control energy expenditure, thermogenesis and weight loss. However, canonic regulators of leptin secretion, such as insulin and malonyl CoA, do not support these processes. We hypothesize that epiregulin (EREG), a growth factor that is secreted from fibroblasts under thermogenic and cachexia conditions, induces leptin secretion associated with energy dissipation. The effects of EREG on leptin secretion were studied ex vivo, in the intra-abdominal white adipose tissue (iAb WAT) explants, as well as in vivo, in WT mice with diet-induced obesity (DIO) and in ob/ob mice. These mice were pair fed a high-fat diet and treated with intraperitoneal injections of EREG. EREG increased leptin production and secretion in a dose-dependent manner in iAb fat explants via the EGFR/MAPK pathway. After 2 weeks, the plasma leptin concentration was increased by 215% in the EREG-treated group compared to the control DIO group. EREG-treated DIO mice had an increased metabolic rate and core temperature during the active dark cycle and displayed cold-induced thermogenesis. EREG treatment reduced iAb fat mass, the major site of leptin protein production and secretion, but did not reduce the mass of the other fat depots. In the iAb fat, expression of genes supporting mitochondrial oxidation and thermogenesis was increased in EREG-treated mice vs control DIO mice. All metabolic and gene regulation effects of EREG treatment were abolished in leptin-deficient ob/ob mice. Our data revealed a new role of EREG in induction of leptin secretion leading to the energy expenditure state. EREG could be a potential target protein to regulate hypo- and hyperleptinemia, underlying metabolic and immune diseases.

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Role of GPX4-Mediated Ferroptosis in the Sensitivity of Triple Negative Breast Cancer Cells to Gefitinib

10.21203/rs.3.rs-70358/v1 ◽

2020 ◽

Author(s):

Xiang Song ◽

Xinzhao Wang ◽

Zhaoyun Liu ◽

Zhiyong Yu

Keyword(s):

Breast Cancer ◽

Cell Viability ◽

Triple Negative Breast Cancer ◽

Triple Negative ◽

Tumor Model ◽

Tunel Staining ◽

Ki 67 ◽

In Vivo Experiments ◽

Related Proteins

Abstract Background: Gefitinib exhibits antitumor activity in the patients with breast cancer, but the resistance to gefitinib in triple negative breast cancer (TNBC) is a new concern. Glutathione peroxidase 4 (GPX4) is a leading regulator of ferroptosis, which is of importance for the survival of TNBC cells. This study investigated GPX4-mediated ferroptosis in gefitinib sensitivity in TNBC.Methods: Gefitinib resistant TNBC cells MDA-MB-231/Gef and HS578T/Gef were constructed, and treated with lentivirus sh-GPX4 and ferroptosis inhibitor ferrostatin-1. GPX4 expression, cell viability and apoptosis were detected. Malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS) levels were evaluated. The levels of ferroptosis-related proteins ACSL4, PTGS2, NOX1 and FTH1 were detected. Subcutaneous tumor model was established in nude mice, and gefitinib was intraperitoneally injected. Apoptosis was detected by TUNEL staining and Ki-67 expression was detected by immunohistochemistry.Results: GPX4 was increased in gefitinib-resistant cells. After silencing GPX4, the inhibition rate of cell viability increased, the limitation of colony formation ability reduced, apoptosis rate increased, and the sensitivity of cells to gefitinib was improved. After silencing GPX4, MDA level and ROS production were significantly increased, while GSH level was decreased. Silencing GPX4 promoted ferroptosis. After inhibition of ferroptosis by ferrostatin-1, it revealed that inhibition of GPX4 promoted gefitinib sensitivity by promoting cell ferroptosis. In vivo experiments also showed that inhibition of GPX4 enhanced the anticancer effect of gefitinib through promoting ferroptosis.Conclusion: Inhibition of GPX4 stimulated ferroptosis and thus enhanced TNBC cell sensitivity to gefitinib.

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Association of bevacizumab-free interval with efficacy of anti-EGFR therapy in patients with metastatic colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.3_suppl.610 ◽

2014 ◽

Vol 32 (3_suppl) ◽

pp. 610-610

Author(s):

Azusa Komori ◽

Hiroya Taniguchi ◽

Yukiya Narita ◽

Shiori Uegaki ◽

Sohhei Nitta ◽

...

Keyword(s):

Colorectal Cancer ◽

Metastatic Colorectal Cancer ◽

Short Interval ◽

Progression Free Survival ◽

Patient Characteristics ◽

Free Interval ◽

Significant Difference ◽

Group A ◽

Group B

610 Background: Both bevacizumab (Bev) and anti-EGFR agents are sequentially used for metastatic colorectal cancer (mCRC). Some basic studies reported the interaction between Bev and anti-EGFR agents in vivo. Therefore we hypothesized the shorter Bev-free interval may lead to poor outcome of anti-EGFR therapy. The aim of this study is to examine the association of the interval between last Bev administration and initial anti-EGFR agents with efficacy of anti-EGFR therapy. Methods: We retrospectively reviewed consecutive mCRC patients who underwent combination therapy of anti-EGFR agents and irinotecan after failure of fluoropyrimidines, oxaliplatin, irinotecan, and Bev at a single institution. We divided patients in two groups (group A: the interval between Bev and anti-EGFR agents <6M, group B: ≥6M). Results: A total of 114 patients constituted the cohort of analysis. The median age was 63; 78 (68%) patients were male. Most patients (N=100, 88%) were treated with cetuximab, and 14 patients were panitumumab. Seventy-four patients were group A and 40 patients group B, respectively. There was no significant difference in patient characteristics. Response rate was 24.7% in group A, and 50.0% in group B (p=0.0072). The patients in group B have significantly longer progression free survival (4.2 vs. 6.6 M, HR 0.65, 95% CI 0.43- 0.98, p=0.042) and longer overall survival (11.6 vs. 14.3 M, HR 0.62, 95% CI 0.39- 0.97, p=0.038). Conclusions: The short interval (<6M) between last Bev and anti-EGFR agents may interfere with the efficacy of anti-EGFR therapy.

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Effectiveness of Antianxiety Drugs on Postoperative Pain Perception After Implant Placement: An In Vivo Study

Journal of Advanced Oral Research ◽

10.1177/2320206820981485 ◽

2021 ◽

Vol 12 (1) ◽

pp. 144-152

Author(s):

Minal Gopal Tulsani ◽

Dhanraj Ganapathy ◽

Divya Rupawat ◽

Sanjana Devi

Keyword(s):

Postoperative Pain ◽

Analysis Of Variance ◽

Pain Perception ◽

Anxiety Level ◽

In Vivo Study ◽

Implant Placement ◽

Significant Difference ◽

Group A ◽

Group B

Aim: To evaluate the effectiveness of midazolam and zolpidem on postoperative pain perception in patients undergoing implant placement. Materials and Methods: In the present in vivo study 60 patients undergoing implant placement were selected based on the inclusion criteria framed and were randomly allocated using sequentially numbered, opaque, and sealed envelope (SNOSE) method into 3 groups with 20 patients each after obtaining informed consent. Group A was the control group, Group B received midazolam 7.5 mg 30 minutes before the procedure. Group C received zolpidem 5 mg 30 minutes before the procedure. The anxiety level of patients was recorded using the Corah scale and postoperative pain was recorded after 2 hours of implant placement using the VAS scale. Statistical analysis was done using analysis of variance (ANOVA), one-way multivariate analysis of variance (one-way MANOVA), and then Tukey’s Honestly Significant Difference (HSD) test for comparison among groups at the 0.05 level of significance. Results: Group A had a mean anxiety level of 16 ± 1.451, Group B had a mean anxiety level of 11.2 ± 2.858, and Group C had a mean anxiety level of 13 ± 2.9019 and a statistically significant difference between the groups was observed ( P < .05). The mean for the postoperative pain perception for Group A was 6.8 ± 1.1965, for Group B was 3.8 ± 1.3611, and Group C was 5 ± 1.451 and a statistically significant difference between the groups was observed ( P < .05). Conclusion: This study concluded that both midazolam and zolpidem significantly reduced anxiety levels and postoperative pain in patients undergoing implant placement.

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