Parasympathetic brainstem origin sites of triggered external and internal carotid vasodilation are not distributed topographically in the rat.

Cranial parasympathetic activation produces vasodilation in the head and neck region, but little is known about its central and peripheral mechanisms. This study was conducted to examine whether external and internal carotid-vasodilation origin sites triggered by chemical stimulation are distributed topographically in the parasympathetic brainstems of anesthetized rats, and to examine the effects of peripheral receptors on vasodilation. Microinjection of the neuromodulator candidate l-cysteine revealed that external and internal carotid vasodilation-triggering sites were distributed non-topographically along the full extent of the parasympathetic parvocellular reticular formation (PcRt). Intravenous injection of a muscarinic blocker and a nitric oxide synthase inhibitor abolished external carotid vasodilation, suggesting the peripheral involvement of muscarinic and nitric oxide receptors. Further work is needed to fully understand the PcRt mechanisms underlying timely and appropriate vasodilation to support various cranial functions.

An origin of carotid vasodilation extends along the full extent of the parasympathetic parvicellular reticular region in the rat brainstem.

During grooming in rats, cranial parasympathetic activation leads to increased carotid artery blood flow, but the brainstem origin of this vasodilation signal is poorly understood. The aim of this study was to map brainstem sites wherein chemical stimulation with l-cysteine, an ionotropic excitatory amino acid receptor activator, can trigger carotid vasodilation in anesthetized intact and superior cervical sympathectomized (SCD) rats. The right side of the brainstem was accessed ventrally; arterial blood pressure and right carotid artery flow resistance were monitored. Employing microinjections of l-cysteine into the ipsilateral brainstem, vasodilation triggering sites were identified dorsal and rostral to the rostral ventrolateral medulla (RVLM) presympathetic pressor area, forming a parasagittal plate-like response zone that varied from rat to rat. In SCD rats (N = 6), we observed a vasodilation zone in a region that included the RVLM pressor area, suggesting a parasympathetic origin. A similar zone was identified in intact rats (N = 8), except that it did not include part of the RVLM pressor area. Dye injected into vasodilator sites labelled the rostral part of the parvicellular reticular formation (PcRt), consistent with a cranial parasympathetic origin. Employing mapping with L-cysteine, a putative neuromodulator, this study provides the first functional demonstration of a carotid vasodilation area that extends along the full extent of the parasympathetic rostral PcRt region in rats.

Topographic carotid vasoconstriction in the rostral ventrolateral medulla of rats.

The vascular beds of various cranial tissues receive common carotid flow, which contributes to blood flow redistribution associated with animal behaviors such as grooming, but the medullary autonomic regulation of carotid flow resistance (CAR) is poorly understood. This study is the first to examine the response sites of CAR in the rat rostral ventrolateral medulla (RVLM) presympathetic area to chemical stimulation by the ionotropic excitatory amino acid receptors activator L-cysteine. Arterial blood pressure and CAR were monitored in anesthetized rats which had a cranial window constructed above the ventral medulla. Mapping of L-cysteine microinjection in eight rats showed carotid vasoconstriction in the caudal part alone within the RVLM pressor area, which included contributions from other vascular beds, indicating localized topographic carotid vasoconstriction. Additional testing was performed on four types of denervated rats. A similar response map was obtained in six rats that received minimal lesions during surgery as well as in 10 rats with severed internal or external carotid nerves. However, the remaining three minimally lesioned rats showed extensive vasoconstriction of the RVLM pressor area including the rostral part, indicating lack of a topographic response. The topographic response of most rats might be state-dependent. Seven rats with complete cervical denervation showed no carotid vasoconstrictor response in the RVLM pressor area, indicating cervical sympathetic mediation of the responses. The topographic carotid vasoconstriction in response to L-cysteine may suggest differential roles of presympathetic neurons in the rostral and caudal parts of the RVLM in sympathetic carotid flow regulation.

Muscle vasodilator response via potential adrenaline secretion to L-cysteine microinjected in rostral ventrolateral medulla of rats.

We previously found that the thiol amino acid L-cysteine microinjected into rat medullary autonomic areas produces changes in arterial blood pressure (AP) via ionotropic excitatory amino acid receptors (iEAAr), but its effect on vascular beds is still unknown. Rostral ventrolateral medulla (RVLM) pressor area includes adrenal and lumbar presympathetic neurons which activation could cause opposite muscle vascular responses: vasodilation versus vasoconstriction. However, there are few data on the vasodilator response in rats. Ionotropic EAAr activation alone with L-cysteine may be effective to differentiate RVLM sites for those opposite responses. To test it, muscle blood flow responses to L-cysteine were mapped in the ventrolateral medulla (VLM) of rats. In anesthetized rats with a cranial window above the VLM, hindquarter flow (HQF) and AP were monitored, providing hindquarter resistance (HQR) by dividing AP by HQF. L-Cysteine mapping in VLM including caudal depressor vasodilator area defined with L-glutamate showed HQR responses in parallel to AP responses, suggesting the importance of iEAAr in muscle vascular regulation. Microinjections of L-cysteine into RVLM succeeded to detect sites of slower muscle vasodilation and blockade of peripheral ß-adrenoceptors abolished this response, indicating potential adrenaline secretion. Although there was no functional topography, the iEAAr activation alone with L-cysteine can differentiate sites of muscle vasodilation from vasoconstriction in rat RVLM. The neuromodulator candidate L-cysteine is a useful tool when chemical stimulation of iEAAr is required.

Central cardiovascular actions of L-homocysteine microinjected into ventrolateral medullary autonomic areas of the rat.

Elevated L-homocysteine concentrations in the plasma and cerebrospinal fluid are related to cardiovascular and neuronal diseases, and could contribute to disease development. However, the central cardiovascular actions of L-homocysteine in two important autonomic regulating areas remain unknown: the rostral ventrolateral medulla (RVLM), including pre-sympathetic neurons, and the caudal ventrolateral medulla (CVLM), including interneurons projecting to pre-sympathetic neurons in the RVLM. Therefore, the aim of the current study was to examine the influence of L-homocysteine microinjected into the RVLM and CVLM areas on changes in arterial blood pressure (ABP) and heart rate (HR) of anesthetized rats, as well as the influence of ionotropic excitatory amino acid (iEAA) receptors on the central actions of L-homocysteine. L-Homocysteine solutions were microinjected into the RVLM and CVLM, which were defined according to pressor and depressor responses to L-glutamate microinjections, respectively. ABP and HR increased in the RVLM and decreased in the CVLM after microinjection with L-homocysteine, similar to L-glutamate, in a dose-dependent manner, suggesting mediation of EAA receptors. Prior microinjection of the N-methyl-D-aspartate (NMDA) iEAA receptor antagonist MK801, but not the non-NMDA receptor antagonist CNQX, abolished the observed responses to L-homocysteine in both the RVLM and CVLM. These results indicate the central cardiovascular actions of L-homocysteine via MK801-sensitive receptors of the medullary autonomic neurons in the rat RVLM and CVLM. It remains unknown if the central cardiovascular actions are related to cardiovascular diseases after endogenously and locally augmented L-homocysteine production by disordered metabolism. Further studies on functional significance of L-homocysteine may provide some clue to understand its toxic mechanism.

L-Cysteine and L-AP4 microinjections in the rat caudal ventrolateral medulla decrease arterial blood pressure.

The thiol amino acid L-cysteine increases arterial blood pressure (ABP) when injected into the cerebrospinal fluid space in conscious rats, indicating a pressor response to centrally acting L-cysteine. A prior synaptic membrane binding assay suggests that L-cysteine has a strong affinity for the L-2-amino-4-phosphonobutyric acid (L-AP4) binding site. The central action of L-cysteine may be vial-AP4 sensitive receptors. The present study investigated cardiovascular responses to L-cysteine and L-ap4 microinjected into the autonomic area of the caudal ventrolateral medulla (CVLM) where inhibitory neurons regulate ABP via pre-sympathetic vasomotor neurons. Both the injection of L-cysteine and L-AP4 in the CVLM sites identified with L-glutamate produced the same depressor and bradycardic responses in urethane-anesthetized rats. Neither a prior antagonist microinjection of MK801 for the N-methyl-D-aspartate (NMDA) receptor nor CNQX for the non-NMDA receptor attenuated the responses to L-cysteine, but the combination of the two receptor blocking with an additional prior injection abolished the response. In contrast, either receptor blockade alone abolished the response to L-AP4, indicating distinct mechanisms between responses to L-cysteine and L-AP4 in the CVLM. The results indicate that the CVLM is a central active site for L-cysteine's cardiovascular response. Central L-cysteine's action could be independent of the L-AP4 sensitive receptors. Cardiovascular regulation may involve endogenous L-cysteine in the CVLM. Further multidisciplinary examinations are required to elaborate on L-cysteine's functional roles in the CVLM.

Cardiovascular actions of L-cysteine and L-cysteine sulfinic acid in the nucleus tractus solitarius of the rat.

The sulfur-containing excitatory amino acid (EAA) L-cysteine sulfinic acid (CSA), a neurotransmitter candidate, is endogenously synthesized from L-cysteine (Cys). Exogenous Cys administration into the brain produces cardiovascular effects; these effects likely occur via synaptic stimulation of central nervous system (CNS) neurons that regulate peripheral cardiovascular function. However, the cardiovascular responses produced by CNS Cys administration could result from CSA biosynthesized in synapse. The present study examined the role of CSA in Cys-induced cardiovascular responses within the nucleus tractus solitarius (NTS) of anesthetized rats. The NTS receives input from various visceral afferents that gate autonomic reflexes, including cardiovascular reflexes. Within the NTS, both Cys and CSA microinjections produced decrease responses in arterial blood pressure and heart rate that were similar to those produced by L-glutamate. Co-injection of the ionotropic EAA receptor antagonist kynurenic acid abolished Cys-, but not CSA-, induced cardiovascular responses. This finding suggests that only Cys-induced cardiovascular responses are mediated by kynurenate-sensitive receptors. This study provides the first demonstration that Cys- and CSA-induced cardiovascular responses occur via different mechanisms in the NTS of rats. Further, this study also indicates that Cys-induced cardiovascular responses do not occur via CSA. Thus, within the NTS, endogenous Cys and/or CSA might be involved in cardiovascular regulation.

Functional cardiovascular action of L-cysteine microinjected into pressor sites of the rostral ventrolateral medulla of the rat.

The endogenous sulfur-containing amino acid L-cysteine injected into the cerebrospinal fluid space of the cisterna magna increases arterial blood pressure (ABP) and heart rate (HR) in the freely moving rat. The present study examined (1) cardiovascular responses to L-cysteine microinjected into the rostral ventrolateral medulla (RVLM), where a group of neurons regulate activities of cardiovascular sympathetic neurons and (2) involvement of ionotropic excitatory amino acid (iEAA) receptors in response. In the RVLM of urethane-anesthetized rats accessed ventrally and identified with pressor responses to L-glutamate (10 mM, 34 nl), microinjections of L-cysteine increased ABP and HR dose dependently (3-100 mM, 34 nl). The cardiovascular responses to L-cysteine (30 mM) were not attenuated by a prior injection of either antagonist alone, MK801 (20 mM, 68 nl) for the NMDA type of iEAA receptors, or CNQX (2 mM) for the non-NMDA type. However, inhibition of both NMDA and non-NMDA receptors with additional prior injection of either antagonist completely blocked those responses to L-cysteine. The results indicate that L-cysteine has functional cardiovascular action in the RVLM of the anesthetized rat, and the responses to L-cysteine involve both NMDA and non-NMDA receptors albeit in a mutually exclusive parallel fashion. The findings may suggest endogenous roles of L-cysteine indirectly via iEAA receptors in the neuronal network of the RVLM for cardiovascular regulation in physiological and pathological situations.

Pressor response to L-cysteine injected into the cisterna magna of conscious rats involves recruitment of hypothalamic vasopressinergic neurons.

The sulfur-containing non-essential amino acid L-cysteine injected into the cisterna magna of adult conscious rats produces an increase in blood pressure. The present study examined if the pressor response to L-cysteine is stereospecific and involves recruitment of hypothalamic vasopressinergic neurons and medullary noradrenergic A1 neurons. Intracisternally injected D-cysteine produced no cardiovascular changes, while L-cysteine produced hypertension and tachycardia in freely moving rats, indicating the stereospecific hemodynamic actions of L-cysteine via the brain. The double labeling immunohistochemistry combined with c-Fos detection as a marker of neuronal activation revealed significantly higher numbers of c-Fos-positive vasopressinergic neurons both in the supraoptic and paraventricular nuclei and tyrosine hydroxylase containing medullary A1 neurons, of L-cysteine-injected rats than those injected with D-cysteine as iso-osmotic control. The results indicate that the cardiovascular responses to intracisternal injection of L-cysteine in the conscious rat are stereospecific and include recruitment of hypothalamic vasopressinergic neurons both in the supraoptic and paraventricular nuclei, as well as of medullary A1 neurons. The findings may suggest a potential function of L-cysteine as an extracellular signal such as neuromodulators in central regulation of blood pressure.

Amino acids that centrally influence blood pressure and regional blood flow in conscious rats.

Functional roles of amino acids have increasingly become the focus of research. This paper summarizes amino acids that influence cardiovascular system via the brain of conscious rats. This paper firstly describes why amino acids are selected and outlines how the brain regulates blood pressure and regional blood flow. This section includes a concise history of amino acid neurotransmitters in cardiovascular research and summarizes brain areas where chemical stimulations produce blood pressure changes mainly in anesthetized animals. This is followed by comments about findings regarding several newly examined amino acids with intracisternal stimulation in conscious rats that produce changes in blood pressure. The same pressor or depressor response to central amino acid stimulations can be produced by distinct mechanisms at central and peripheral levels, which will be briefly explained. Thereafter, cardiovascular actions of some of amino acids at the mechanism level will be discussed based upon findings of pharmacological and regional blood flow measurements. Several examined amino acids in addition to the established neurotransmitter amino acids appear to differentially activate brain structures to produce changes in blood pressure and regional blood flows. They may have physiological roles in the healthy brain, but pathological roles in the brain with cerebral vascular diseases such as stroke where the blood-brain barrier is broken.

Intracisternally injected L-proline activates hypothalamic supraoptic, but not paraventricular, vasopressin-expressing neurons in conscious rats.

When injected into specific rat brain regions, the neurotransmitter candidate L-proline produces various cardiovascular changes through ionotropic excitatory amino acid receptors. The present study used an immunohistochemical double-labeling approach to determine whether intracisternally injected L-proline in freely moving rats, which increases blood pressure, activates hypothalamic vasopressin-expressing neurons and ventral medullary tyrosine-hydroxylase- (TH-) containing neurons. Following injection of L-proline, the number of activated hypothalamic neurons that coexpressed vasopressin and c-Fos was much greater in the supraoptic nucleus (SON) than in the paraventricular nucleus (PVN) of rats with increased blood pressure. The number of activated TH-containing neurons was significantly greater following L-proline treatment than following control injections of artificial cerebrospinal fluid (ACSF). These results clearly demonstrate that intracisternally injected L-proline activates hypothalamic supraoptic, but not paraventricular, vasopressin-expressing neurons and medullary TH-containing (A1/C1) neurons in freely moving rats.

The mapped pattern of kainate on blood pressure responses is similar to that of L-proline in the ventrolateral medulla of the rat.

Kainate is an excitatory amino acid receptor agonist with a structure similar to the amino acid L-proline. Our previous studies demonstrated that microinjections of L-proline into the ventrolateral medulla (VLM) of the rat induce a mapped pattern of blood pressure responses distinct from L-glutamate, and the depressor response to L-proline in the caudal VLM (CVLM) is abolished by the kainate/AMPA receptor antagonist CNQX. The present study investigated whether kainate produces the L-proline-mapped pattern of responses in the VLM, compared with the pattern by AMPA. Kainate is known to activate AMPA receptors at higher concentrations. Therefore, responses to kainate were investigated at a low concentration. Microinjections of AMPA or NMDA showed the pattern of the L-glutamate-type; a pressor response in the rostral VLM and caudal pressor area (CPA) and a depressor response in the CVLM. Microinjections of kainate showed depressor responses in the CVLM but minor pressor responses in the rostral VLM, suggesting the same responses to L-proline. However, the response sites in the CPA did not enable us to clearly determine the L-proline-type. Further trials at sites defined by a pressor response to L-glutamate in the CPA, successive injections of L-proline and kainate produced no response, indicating that L-glutamate responding neurons in the CPA are not sensitive to L-proline and kainate. These results suggest that kainate stimulation in the VLM produces a mapped pattern of ABP responses similar to the mapped pattern with L-proline. Kainate receptors could therefore be involved in the depressor response to L-proline in the medulla.

Antiproliferative effects of 5-fluorouracil and interferon-alpha in combination on a hepatocellular carcinoma cell line in vitro and in vivo.

BACKGROUND AND AIM: We investigated the antiproliferative effects of interferon-alpha (IFN-alpha) and 5-fluorouracil (5-FU) in combination on a hepatocellular carcinoma (HCC) cell line. METHOD: In the in vitro study, IFN-alpha and/or 5-FU was added to the culture of the poorly differentiated-type HCC cell line, HAK-1B, and their antiproliferative effects and additional or synergic effects in combination treatment were examined. In the in vivo study, HAK-1B cells were transplanted into nude mice and the changes in tumor volume and weight, apoptosis, BrdU and cyclin A positive cells, and artery-like blood vessels were investigated. Expressions of angiogenesis factors and IFN-alpha receptor (IFNAR-2) were examined in the developed tumors. RESULTS: In vitro growth of HAK-1B cells was suppressed dose-dependently to 5-FU, but the addition of IFN-alpha did not induce additional or synergic effects. In vivo growth in terms of tumor diameter and weight was suppressed at most in the IFN-alpha + 5-FU (combination) group, that is, the tumor volume became 29.3% and the tumor weight became 54.7% of the control. In the combination group, numbers of BrdU-positive S-phase cells and cyclin A positive cells increased together with the increase in apoptotic cells, but there was no significant relation between the tumor shrinkage effects and angiogenesis factors or artery-like blood vessels. In the combination group, INFAR-2 decreased significantly in comparison to the other groups. CONCLUSION: The synergic growth-suppression effects in the current in vivo study using the combination treatment are attributable to the enhanced induction of S-phase arrest and of apoptosis.

Immunohistochemical evidence for the localization of neurons containing the putative transmitter L-proline in rat brain.

We examined whether there are the neurotransmitter candidate amino acid L-proline containing neurons localized in the rat brain. Antibodies against L-proline conjugated with rabbit serum albumin were raised in a rabbit and purified with affinity chromatography. Strong L-proline-like immunoreactivity was confined to several groups of neurons in the arcuate nucleus (n) and supraoptic n in the hypothalamus and area postrema. The brainstem had markedly stained fibers in the medial longitudinal fasciculus and localized neuronal cell body labeling in the red n, mesencephalic trigeminal n, lateral reticular n, raphe obscurus n, solitary n, compact ambiguus n, motor trigeminal n and n of trapezoid body. Our findings are consistent with the hypothesis that L-proline may function as a neurotransmitter or neuromodulator in the brain.

Depressor responses to L-proline microinjected into the rat ventrolateral medulla are mediated by ionotropic excitatory amino acid receptors.

The essential amino acid L-proline produces a depressor response when microinjected into the caudal ventrolateral medulla (CVLM) of anesthetized rats. L-proline may activate some excitatory amino acid (EAA) receptors. The present study tested this hypothesis by investigating the effects of two ionotropic excitatory amino acid receptor antagonists on the depressor response to L-proline in the CVLM: the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate receptor-selective antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and the N-methyl-D-aspartate (NMDA) receptor-selective antagonist MK801. Urethane-anesthetized rats received arterial catheters and their ventrolateral medulla surface was exposed. Injections of the antagonists CNQX and MK801 (2 mM, 68 nl in each case) into the CVLM completely blocked depressor responses to subsequent administration of AMPA (2 pmol/34 nl) and NMDA (2 pmol/34 nl), respectively. The depressor response to L-proline (3.4 nmol/34 nl) was strongly inhibited by prior injection of CNQX (2 mM, 68 nl) and significantly attenuated by prior injection of a high dose (20 mM, 68 nl), but not a low dose (2 mM, 68 nl), of MK801. The results indicate that the depressor response to L-proline in the CVLM includes mechanisms of ionotropic excitatory amino acid receptors.

Expression of matrix metalloproteinases (MMPs) in cultured hepatocellular carcinoma (HCC) cells and surgically resected HCC tissues.

Matrix metalloproteinases (MMPs) relate to the growth and infiltration of cancer cells, but the frequency and amount of their expression are not yet fully examined in hepatocellular carcinoma. Expression of MMPs (MMP-2, MMP-7, MMP-9, MT1-MMP, MT2-MMP, MT3-MMP) and tissue inhibitors of metalloproteinase (TIMP: TIMP-1, TIMP-2) was investigated on cultured hepatocellular carcinoma (HCC) cells and surgically resected HCC tissues. The cultured cells and tissues expressed MMPs and TIMPs at various degrees, and high expression was observed for MMP-2, MMP-9, MT1-MMP and TIMP-2. Expression of MMP-7, MT2-MMP and TIMP-1 was found at a low frequency and a low amount in both the cells and the tissues. MMP-2 was expressed in various cells: HCC cells, vascular wall and sinusoidal endothelial cells in the cancer area of surgically resected tissues; and hepatocytes, bile duct cells, vascular wall, macrophages and Kupffer cells in the non-cancerous area. MMPs and TIMPs were expressed at a relatively high frequency in hepatocytes of the cancerous area and surrounding non-cancerous area as well as in the other cells and tissues. MMPs and TIMPs may be involved in the progression of hepatocellular carcinoma including the infiltration of cancer cells.

Vasodilatation of the hindquarters induced by antagonism of GABA(A) receptors in the freely moving rat.

An intracisternal injection of the GABA(A) receptor antagonist bicuculline into conscious rats increased arterial pressure and decreased hindquarter resistance. Propranolol attenuated only the resistance response, but ganglionic block abolished both responses. These findings suggest that central GABA(A) receptor blockade induces an autonomic pressor action with hindquarter vasodilatation.

Antiproliferative effects of interferon-alphaCon1 on ovarian clear cell adenocarcinoma in vitro and in vivo.

PURPOSE: We examined the antiproliferative effect of IFN-alphaCon1 and its mechanism on ovarian clear cell adenocarcinoma in vitro and in vivo. EXPERIMENTAL DESIGN: (a) The effects of IFN-alphaCon1 on growth, morphology, cell cycle, and type I IFN-alpha receptor (IFNAR-2) expression were examined on two ovarian clear cell adenocarcinoma cell lines (KOC-5C and KOC-7C) in vitro. (b) KOC-5C or KOC-7C cells were transplanted into nude mice, and changes in tumor volume, tumor weight, apoptosis, necrosis, and microvessel density were investigated. The expression of angiogenesis factors was examined in the serum and the developed tumors. RESULTS: Both cell lines expressed IFNAR-2 mRNA, but its protein was detected only in KOC-7C. In KOC-7C cells, antiproliferative effects were observed in a time- and dose-dependent manner and cell division was blocked at the S phase. The KOC-7C tumors showed decreases in tumor volume and weight; a decreasing tendency in basic fibroblast growth factor (bFGF), vascular endothelial growth factor, and interleukin (IL)-8 protein expression in the tumor; a significant decrease in bFGF and IL-8 protein expression in the serum, and of microvessel density; and significant increase in apoptosis and necrosis in the tumor. In the KOC-5C tumors, these in vitro and in vivo changes were not apparent, and the antiproliferative effects of IFN-alphaCon1 were not obvious. CONCLUSIONS: IFN-alphaCon1 suppresses tumor proliferation by inducing apoptosis, blocking the cell cycle, and inhibiting tumor angiogenesis. Our findings show that the clinical efficacy of IFN-alphaCon1 can be predicted by examining IFNAR-2 expression on tumor cells, and the efficacy of IFN-alphaCon1 treatment can be evaluated by measuring serum bFGF and IL-8 levels.

Interferon-alphaCon1 suppresses proliferation of liver cancer cell lines in vitro and in vivo.

BACKGROUND/AIMS: We investigated the effects of consensus interferon (IFN-alphaCon1), a nonnaturally occurring type I interferon with higher specific activity than other type I IFNs, on the growth of human liver cancer cells. METHODS: The effect of IFN-alphaCon1 on the proliferation of 13 liver cancer cell lines was investigated in vitro. Hepatocellular carcinoma (HCC) cells (KIM-1 and HAK-1B) were transplanted subcutaneously into the back of nude mice, then IFN-alphaCon1 was subcutaneously administered to the mice once a day for 2 weeks, and tumor volume and histology were examined. RESULTS: IFN-alphaCon1 expressed a dose-dependent growth inhibitory effect in all cell lines in vitro. KIM-1 tumor volume in mice that received 0.01 microg (10(4)IU)/mouse/day of IFN-alphaCon1 (similar to the clinical dose for chronic hepatitis C) was 62% of the control, 0.1microg/mouse/day resulted in 26%, and 1 microg/mouse/day resulted in 10%. HAK-1B tumor volume under the same treatment was 61, 24 and 0% of the control, respectively. The number of apoptotic cells significantly increased and the number of blood vessels significantly decreased with the increase in IFN-alphaCon1 dose. CONCLUSIONS: IFN-alphaCon1 suppressed HCC growth in nude mice. These data indicate the potential clinical application of IFN-alphaCon1 in the prevention and treatment of HCC.

Co-expression of Y box-binding protein-1 and P-glycoprotein as a prognostic marker for survival in epithelial ovarian cancer.

OBJECTIVES: This study aimed to observe the expressions of Y box-binding protein-1 (YB-1) and P-glycoprotein (P-gp) in primary ovarian tumor and to determine whether they act as biomarkers for survival in epithelial ovarian cancer. METHODS: The expressions of YB-1 and P-gp were examined immunohistochemically in 59 patients who were treated from 1997 to 2000 at Kurume University Hospital. Samples were paraffin-embedded primary ovarian cancer tissue taken from the surgical specimens. RESULTS: Of the 59 primary ovarian tumors examined, 32 (54.2%) and 18 (30.5%) were positive for YB-1 and P-gp, respectively. Co-expression of these two proteins was observed in 20.3% (12/59) cases. Patients showing this co-expression had a worse 3-year survival than those without co-expression (40.0% vs. 73.1%, P = 0.0447). This co-expression significantly correlated with poor prognosis according to multivariate analysis (P = 0.0007). CONCLUSION: Co-expression of YB-1 and P-gp emerged as a promising relevant biomarker for unfavorable prognosis in ovarian cancer.