NF-κB signaling regulates cell-autonomous regulation of CXCL10 in breast cancer 4T1 cells

The chemokine CXCL10 and its receptor CXCR3 play a role in breast cancer metastasis to bone and osteoclast activation. However, the mechanism of CXCL10/CXCR3-induced intracellular signaling has not been fully investigated. To evaluate CXCL10-induced cellular events in the mouse breast cancer cell line 4T1, we developed a new synthetic CXCR3 antagonist JN-2. In this study, we observed that secretion of CXCL10 in the supernatant of 4T1 cells was gradually increased during cell growth. JN-2 inhibited basal and CXCL10-induced CXCL10 expression and cell motility in 4T1 cells. Treatment of 4T1 cells with CXCL10 increased the expression of P65, a subunit of the NF-κB pathway, via activation of the NF-κB transcriptional activity. Ectopic overexpression of P65 increased CXCL10 secretion and blunted JN-2-induced suppression of CXCL10 secretion, whereas overexpression of IκBα suppressed CXCL10 secretion. These results indicate that the CXCL10/CXCR3 axis creates a positive feedback loop through the canonical NF-κB signaling pathway in 4T1 cells. In addition, treatment of osteoblasts with conditioned medium from JN-2-treated 4T1 cells inhibited the expression of RANKL, a crucial cytokine for osteoclast differentiation, which resulted in an inhibitory effect on osteoclast differentiation in the co-culture system of bone marrow-derived macrophages and osteoblasts. Direct intrafemoral injection of 4T1 cells induced severe bone destruction; however, this effect was suppressed by the CXCR3 antagonist via downregulation of P65 expression in an animal model. Collectively, these results suggest that the CXCL10/CXCR3-mediated NF-κB signaling pathway plays a role in the control of autonomous regulation of CXCL10 and malignant tumor properties in breast cancer 4T1 cells.

alpha-Tocopheryl Succinate Inhibits Osteoclast Formation by Suppressing Receptor Activator of Nuclear Factor-kappaB Ligand (RANKL) Expression and Bone Resorption

OBJECTIVE: Osteoclasts are bone-resorbing multinucleated cells derived from the monocyte/macrophage lineage during normal and pathological bone turnover. Recently, several studies revealed that alpha-tocopheryl succinate (alphaTP-suc) have demonstrated potent anti-cancer activities in vitro and in vivo. However, the effects of alphaTP-suc on osteoclast formation and bone resorption remain unknown. Thus, in this study, we examined the effects of alphaTP-suc on osteoclast differentiation and bone resorbing activity in inflammatory bone loss model. METHODS: Osteoclast differentiation assay was performed by cocultures of mouse bone marrow cells and calvarial osteoblasts in culture media including interleukin-1 (IL-1). Osteoclasts were stained for tartrate-resistant acid phosphatase (TRAP). The level of receptor activator of nuclear factor-kappaB ligand (RANKL) mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). ICR mice were administered an intraperitoneal injections of alphaTP-suc or dimethyl sulfoxide (DMSO) 1 day before the implantation of a freeze-dried collagen sponge loaded with phosphate-buffered saline (PBS) or IL-1 over the calvariae and every other day for 7 days. The whole calvariae were obtained and analyzed by micro-computed tomography (CT) scanning, and stained for TRAP. RESULTS: alphaTP-suc inhibits osteoclast formation in cocultures stimulated by IL-1 and decreased the level of expression of RANKL mRNA in osteoblasts. In addition, administered intraperitoneal injections of alphaTP-suc prevented IL-1-mediated osteoclast formation and bone loss in vivo. CONCLUSION: Our findings suggest that alphaTP-suc may have therapeutic value for treating and preventing bone-resorptive diseases, such as osteoporosis.

Antinociceptive Effects of Intrathecal Melatonin on Formalin: and Thermal-induced Pain in Rats / 대한통증학회지

BACKGROUND: It has been known that melatonin is involved in the modulation of nociceptive transmission. However, the effect of melatonin administered spinally has not been examined. Therefore, we examined the effect of melatonin on the formalin-induced or thermal-induced nociception at the spinal level. METHODS: Intrathecal catheter was inserted into the subarachnoid space of male Sprague-Dawley rats. Pain was assessed by formalin test (induced by injection of 50microliter of a 5% formalin solution to the hindpaw) or Hot-Box test (induced by radiant heat application to the hindpaw). The effect of intrathecal melatonin was examined on flinching behavior in the formalin test or withdrawal response in Hot-Box test. RESULTS: Intrathecal melatonin produced a limited, but dose-dependent reduction of the flinching response during phase 1 and 2 in the formalin test. In addition, melatonin delivered at evening also decreased the flinching response in both phases of the formalin test. Melatonin restrictively increased the withdrawal latency in Hot-Box test. CONCLUSIONS: These results suggest that melatonin is active against the formalin- and thermal-induced nocicpetion at the spinal level, but the effect is limited.

Assessment for the Role of Serotonin Receptor Subtype 3 for the Analgesic Action of Morphine at the Spinal Level / 대한통증학회지

BACKGROUND: Serotonin 3 receptor is involved in the modulation of nociceptive transmission in the spinal cord. The serotonin 3 receptor antagonist has been used for the management of opioid-induced nausea and vomiting. The aim of this study was to examine whether the analgesic effect of morphine is antagonized by serotonin 3 receptor antagonists at the spinal level. METHODS: Rats were implanted with lumbar intrathecal catheters. For nociception, a formalin solution (5%, 50microliter) was injected into the hind paw of male Sprague-Dawley rats. To determine whether the effect of intrathecal morphine was mediated via serotonin 3 receptors, serotonin 3 receptor antagonists were intrathecally administered 10 min prior to the morphine delivery. Following the formalin injection, formalin-induced nociceptive behavior (flinching response) was observed for 60 min. RESULTS: Intrathecal morphine produced a dose-dependent suppression of the flinches in both phases during the formalin test. The analgesic action of morphine was not reversed by serotonin 3 receptor antagonists (LY-278, 584, ondansetron), which had little per se effect on the formalin-induced nociception. CONCLUSIONS: Spinal serotonin 3 receptors may not be involved in the analgesia of morphine on a nociceptive state evoked by a formalin stimulus.

Study for the Antinociceptive Effect and Toxicity of Chronic Intrathecal Infusion of Cannabinoids in Rats / 대한통증학회지

BACKGROUND: Cannabinoids have shown antinociceptive action. The aims of this study were to examine the effect of chronic infusion of a cannabinoids receptors agonist (WIN 55, 212-2) for thermal nociception at the spinal level, and to also observe the development of toxicity. METHODS: Male Sprague-Dawley rats were implanted with lumbar intrathecal catheters with the nociceptive response (withdrawal response latency) determined by exposing the plantar surface of the hindpaw to radiant heat. Initially, the effect of intrathecal WIN 55, 212-2 was evaluated followed by the change in the effect at 1, 2, 3 and 4 weeks after repeated infusion. Finally, the histopathological findings were assessed 1 and 4 weeks following the infusion of WIN 55, 212-2. RESULTS: Intrathecal WIN 55, 212-2 was found to produce a limited antinociception during the thermal test. %MPE of WIN 55, 212-2 at 1, 2, 3, and 4 weeks after infusion was not different from each other. No abnormal pathological findings were observed following a chronic intrathecal infusion of WIN 55, 212-2. CONCLUSIONS: WIN 55, 212-2, a cannabinoids receptors agonist, may be useful in the management of thermal nociception, without changing the effectiveness or causing the toxicity following a chronic infusion at the spinal level.

The Effect of Doxapram Hydrochloride on the Ventilation Responses during General Anesthesia with Volatile Agent by Laryngeal Mask Airway / 대한마취과학회지

BACKGROUND: Inhalation anesthetics are known to depress ventilatory response to hypercapnea. Doxapram hydrochloride is an analeptic drug, which acts as a respiratory stimulant via peripheral and central chemoreceptors. Although the postoperarive infusion of doxapram hydrochloride is known to attenuate the impairment of respiratory function, no report is available on respiratory response to this drug when applied during anesthesia. Therefore, the present study aimed to evaluate the effect of doxapram hydrochloride on respiratory function during anesthesia. METHODS: Sixty adult patients undergoing operation under spontaneous ventilation via laryngeal mask airway (LMA) were randomly categorized into 3 groups: A control group, which received 5% dextrous infusion, and two groups in which patients were infused with doxapram hydrochloride (0.5 or 2 mg/kg/hr) starting 15 min after commencement operation. Anesthesia was maintained with 1 MAC sevoflurane - 4 L N2O - 2 L O2 under spontaneous ventilation via LMA. Tidal volume (VT), respiratory rate (RR), and arterial carbon dioxide tension (PaCO2) were measured just before and 15 min after the induction of anesthesia, 15 min after the start of operation and 15, 30, 45, and 60 min after the start of doxapram hydrochloride infusion. RESULTS: Measured values of RR and PaCO2 were significantly elevated during anesthesia venous those measured just before the induction of anesthesia in all groups. VT was significantly reduced during anesthesia venous just before the induction of anesthesia in all groups. All percent changes of VT, RR and PaCO2 were similar all any measurement times, and showed no significant changes after the infusion of doxapram hydrochloride in all groups. CONCLUSIONS: Intraoperative doxapram hydrochloride treatment did not produce any significant respiratory response improvement during 1 MAC sevoflurane anesthesia.

Influence of Collapse and Re-ventilation of Lung on the Development of Pulmonary Edema / 대한구급학회지

BACKGROUND: This study was to clarify the influence of collapse and re-ventilation of lung on the development of pulmonary edema in rabbit. METHODS: Animals were randomly assigned to one of three groups: Sham group receiving two lung ventilation (n=14), Collapse group receiving collapse of right lung (n=14), Reventilation group receiving collapse of right lung for 3 hours followed by reventilation of collapsed right lung for 3 hours (n=14). The lung of rabbits were ventilated with 50% oxygen through the tracheostomy. Right main bronchus was secured by thoracotomy in all animal. Collapse and reventilation were performed using by bulldog forcep. Mean arterial pressure, heart rate, arterial oxygen tension (PaO2), peripheral blood leukocyte and platelet counts were recorded at 0, 1, 2, 3, 4, 5 and 6 hour after the start of experiment. The wet to dry (W/D) weight ratio of lung, lung injury score and leukocyte counts, percentage of polymorphonuclear leukocyte (PMNL), concentration of albumin, and interleukin-8 (IL-8) in bronchoalveolar lavage fluid (BALF) were measured 6 hour after the start of experiment in both lung. RESULTS: W/D weight ratio of lung, lung injury score and leukocyte counts, percentage of PMNL, concentration of albumin and IL-8 in BALF were significantly increased in both lung of reventilation group. And the degree of increases is more significant in right than left lung. CONCLUSIONS: These findings suggest that reventilation of collapsed lung causes the bilateral pulmonary edema in rabbit mainly by activating neutrophil and IL-8 responses, which may play a central role in non cardiogenic pulmonary edema.

A Case of Nonsecretory Multiple Myeloma / 대한혈액학회지

A case of IgA lambda nonsecretory multiple myeloma in a 66-year-old man was reported. Despite of the osteolytic lesions both protein electrophoresis and protein immunoelectrophoresis of serum and urine of the patient were normal. Bone marrow biopsy at iliac crest showed 8% plasma cells and aspiration cytology of the lesion of rib revealed 73% plasma cells. When examined by immunofluorescence with monospecific antisera the cytoplasm of the immature plasma cell showed predominantly the presence of IgA and lambda chains.

A Case of Recurred Primary Mediastinal Nonseminomatous Germ Cell Tumor Associated with Klinefelters Syndrome / 결핵

Primary mediastinal nonseminomatous germ cell tumor associated with Klinefelter's syndrome is a rare disorder. We experienced a case of recurred primary mediastinal nonseminomatous germ cell tumor developed in a 24-year-old patient with Klinefelter's syndrome. The patient had been treated with surgery and combination chemotherapy under the diagnosis of primary mediastinal nonseminomatous germ cell tumor before. A round mass was found on the right lower lung field in the chest X-ray during follow up. The patient was diagnosed as recurred primary nonseminomatous germ cell tumor and Klinefelter's syndrome through tumor markers, peripheral blood karyotyping, and other tests including hormonal assay and was treated with combination chemotherapy and surgery again. When the patient is diagnosed as primary mediastinal nonseminomatous germ cell tumor, Klinefelter's syndrome and hematologic malignancies should be considered to be associated diseases and vice versa.