Eric V. Yang

Norepinephrine Up-regulates the Expression of Vascular Endothelial Growth Factor, Matrix Metalloproteinase (MMP)-2, and MMP-9 in Nasopharyngeal Carcinoma Tumor Cells

Recent studies using ovarian cancer cells have shown that the catecholamine hormones norepinephrine (norepi) and epinephrine (epi) may influence cancer progression by modulating the expression of matrix metalloproteinases (MMP) and vascular endothelial growth factor (VEGF). The purpose of this study is to determine if the stress hormone norepi can influence the expression of MMP-2, MMP-9, and VEGF in nasopharyngeal carcinoma (NPC) tumors by using three NPC tumor cell lines. The NPC cell lines HONE-1, HNE-1, and CNE-1 were treated with norepi. The effects of norepi on MMP-2, MMP-9, and VEGF synthesis were measured by ELISA; functional MMP activity was measured by the invasive potential of the cells using a membrane invasion culture system whereas functional activity of VEGF was analyzed using a human umbilical vein endothelial cell tube formation assay. Norepi treatment increased MMP-2, MMP-9, and VEGF levels in culture supernatants of HONE-1 cells, which could be inhibited by the beta-blocker propranolol. Norepi induced the invasiveness of all NPC cell lines in a dose-dependent manner, which was blocked by CMT-3, an MMP inhibitor, and propranolol. Norepi stimulated the release of functional angiogenic VEGF by HONE-1 cells as well. Finally, HONE-1 cells were shown to express beta-adrenergic receptors as did seven of seven NPC biopsies examined. The data suggest that catecholamine hormones produced by the sympathetic-adrenal medullary axis may affect NPC tumor progression, in part, through modulation of key angiogenic cytokines.

β-Blockers and Survival among Danish Patients with Malignant Melanoma: A Population-Based Cohort Study

Background: To study whether use of β-blockers increases survival in patients with malignant melanoma because experimental data suggest that catecholamine hormones may be involved in stimulating the aggressiveness of malignant melanoma. Methods: A total of 4,179 patients diagnosed with malignant melanoma in Denmark with a median follow-up of 4.9 years and identified in the Danish Cancer Registry participated. Data on β-blocker use, comorbidity, and survival were obtained from medical and administrative databases. Cox proportional hazards models were used to estimate HRs for all-cause mortality with 95% CIs with adjustment for prognostic factors. Results: A total of 372 (8.9%) patients with malignant melanoma were treated with β-blockers within 90 days of melanoma diagnosis. The median β-blocker duration for exposure within 90 days of melanoma diagnosis, more than 90 days, and no prior exposure was 7.6, 1.4, and 0 years, respectively. The patients receiving β-blockers were older, had more comorbidities, and more cardiovascular and psychotropic drug user than the patients receiving no β-blockers prior to melanoma diagnosis. After adjustment for age and comorbidity index, the HR for melanoma death was 0.87 (95% CI: 0.64–1.20) and for all-cause mortality was 0.81 (95% CI: 0.67–0.97). Conclusion: Increased survival time of patients with melanoma receiving β-blockers suggests that this class of drugs may hold promise in treatment strategy for these patients. Impact: The observations described here suggest that catecholamines may retard melanoma progression and that β-blockers may have unrecognized potential as a therapeutic intervention for melanoma. Cancer Epidemiol Biomarkers Prev; 20(10); 2273–9. ©2011 AACR.

Stress-induced immunomodulation and the implications for health

There are complex bi-directional interactions among the central nervous system (CNS), the endocrine system, and the immune system. Although the mechanisms of this bi-directional communication is not yet fully understood, studies in the field of psychoneuroimmunology (PNI) have shown that stress, through the hypothalamic-pituitary-adrenal (HPA) and the sympathetic-adrenal medullary (SAM) axes, can result in the dysregulation of the immune system. In this review, we discuss human studies and animal models, which focuses on psychological stress emphasizing the implications of these effects on wound healing and infectious diseases.

Stress-related modulation of matrix metalloproteinase expression

Matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs), whose expression can be controlled by cytokines, play a role in extracellular matrix remodeling in physiological and pathological processes. Using a blister chamber wound model on UV-B-exposed human forearm skin, we examined whether stress or mood-associated neuroendocrine alteration is sufficient to modulate MMP and TIMP expression. We did not find evidence that depressive symptoms were reliably associated with modulation of either MMP or TIMP expression. However, we did find that activation of the hypothalamic-pituitary-adrenal (HPA) and sympathetic-adrenal medullary (SAM) axes can modulate levels of MMPs. A positive association between plasma norepinephrine levels and MMP-2 protein levels, and a negative correlation between plasma cortisol levels and MMP-2 levels were found. The data suggest that activation of the HPA and SAM axes, even in individuals within the normal range of depressive symptoms, could mediate MMP levels and wound healing in blister wounds.

Stress-induced immunomodulation: Implications for tumorigenesis

There is some evidence to indicate that psychological factors can affect the incidence and progression of cancer in individuals (Kiecolt-Glaser & Glaser, 1999; Lewis, O Brien, & Barraclough, 2002; Thomas, Pandey, Ramdas, & Nair, 2002). Studies have shown an association between stressful life events and higher incidences of cancer of the lung, breasts, and colon (Courtney, Longnecker, & Peters, 1996; Courtney, Longnecker, Theorell, & Gerhardsson de Verdier, 1993; Forsen, 1991; Horne & Picard, 1979). Although negative behaviors (e.g., cigarette smoking, alcohol use, and unhealthy dietary practices) associated with psychosocial stress may partly explain a higher incidence of cancer and lower survival rates in distressed individuals, it is possible that stress-induced immunosuppression/dysregulation could also be a contributor (Cohen, Kristal, NeumarkSztainaer, Rock, & Neuhouser, 2002; Ellison et al., 2001; Horne & Picard, 1979; Maunsell, Drolet, Brisson, Robert, & Deschenes, 2002; Saxe et al., 2001; Yang & Glaser, 2002). The review ‘‘The Promotion of Tumor Metastasis by Surgery and Stress: Immunological Basis and Implications for Psychoneuroimmunology (PNI)’’ by Ben-Eliyahu discusses how surgical stress could promote tumor progression and presents evidence for how different aspects of surgery (both immunological and non-immunological factors) have been shown to suppress cell-mediated immunity, specifically natural killer (NK) cell activity. Results from both animal and human studies suggest that psychological factors may affect cancer development and progression. Studies in the field of PNI have shown that psychological stress can affect many aspects of immune cell function. In addition, results indicate that the effects of stressors can induce immune dysregulation through modulation by the endocrine system. These interactions are bi-directional (Rabin, 1999). The role of ‘‘immune surveillance’’ in cancer progression has been supported by data showing that subjects who are immunosuppressed have an increased risk for cancer, particularly patients with immunogenic tumors (Herberman & Ortaldo, 1981). Because of their important role in the surveillance of tumors, studies have focused on the effects of psychological stressors on NK cell activity as mediated by neuroendocrine hormones and cytokines (Page, BenEliyahu, & Liebeskind, 1994; Whiteside & Herberman, 1989). It has been suggested that psychological stress brings about the dysregulation of Th1/Th2 cytokines. For instance, observations by Marshall and co-workers (1998) suggest that psychological stress associated with medical student exams can produce a shift in the Th1/ Th2 cytokine balance toward a Th2 response. We have confirmed these data and shown that the chronic stress of caregiving for a dementia patient is also associated with a Th1/Th2 shift (Glaser et al., 2001). This shift could be related to the down-regulation of several immune measures we have observed in studies with healthy medical students. These include decreases in NK cell activity, a decrease in the response of peripheral blood lymphocytes (PBLs) to mitogens, a decrease in production of interferon-gamma (IFN-c) by PBLs in response to stimulation by Concanavalin A (Con A), a decrease in the antibody and virus specific T-cell responses to a hepatitis B vaccination, and changes in the ability of the Brain, Behavior, and Immunity 17 (2003) S37–S40

VEGF is differentially regulated in multiple myeloma-derived cell lines by norepinephrine

Evidence from human and animal studies support the hypothesis that psychological stress can be a co-factor for the initiation and progression of cancer. Recent work from our laboratory and others have shown that the catecholamine hormone, norepinephrine (NE), may influence tumor progression of some solid epithelial tumors including nasopharyngeal carcinoma (NPC) and ovarian cancer by modulating the expression of proangiogenic and pro-metastatic factors, such as vascular endothelial growth factor (VEGF). In this study, we determined whether NE can likewise modulate the expression of VEGF in a lymphoid tumor, multiple myeloma (MM), a cancer of plasma cells. Three MM-derived cell lines, NCI-H929, MM-M1, and FLAM-76, were studied. The presence of beta1- and beta2-adrenergic receptors (ARs) was assessed using Western blotting. Cells were treated with 0, 1, and 10 microM NE for 1, 3, 6, and 24h and the levels of VEGF in culture supernatants were measured by ELISA. Immunoblots of cell lysates revealed the presence of beta1- and beta2-ARs in all three MM-derived cell lines. However, these MM-derived cell lines exhibited varying degrees of NE-dependent regulation of VEGF expression with FLAM-76 (the only IL-6-dependent cell line among the three) exhibiting the most significant stimulation, followed by MM-M1 cells and then NCI-H929. The data suggest that the ability of NE to regulate the expression of VEGF is not limited to solid epithelial tumors and suggests a possible regulatory role of catecholamine stress hormones in MM progression.

Glucocorticoids activate Epstein Barr Virus lytic replication through the upregulation of immediate early BZLF1 gene expression

Psychological stress-associated immune dysregulation has been shown to disrupt the steady-state expression and reactivate latent herpes viruses. One such virus is the Epstein Barr virus (EBV), which is associated with several human malignancies. EBV infects >90% of people living in North America and persists for life in latently infected cells. Although several studies have shown that glucocorticoids (GCs) can directly induce reactivation of the latent virus, the mechanism of stress hormone involvement in the control of EBV gene expression is not well understood. In this study, we tested the hypothesis that GCs can induce the latent EBV genome to lytically replicate through the induction of the EBV immediate early gene BZLF1 which encodes the lytic transactivator protein ZEBRA. We show a dose-dependent upregulation of BZLF1 mRNA expression by hydrocortisone (HC) and dexamethasone (Dex) in Daudi cells, an EBV genome positive Burkitts lymphoma cell line, and Dex-induction of the early gene products BLLF3 (encoding for the EBV dUTPase) and BALF5 (encoding for the EBV DNA polymerase). We show that Daudi cells express glucocorticoid receptors (GR) that mediate Dex-dependent upregulation of BZLF1 mRNA levels. This effect was inhibited by both the glucocorticoid receptor antagonist RU486 and by cycloheximide. The results suggest that GCs, in addition to inducing stress-related immune dysregulation, can mediate latent EBV reactivation through the induction of the BZLF1 gene.

Basal cell carcinoma: Stressful life events and the tumor environment

CONTEXT Child emotional maltreatment can result in lasting immune dysregulation that may be heightened in the context of more recent life stress. Basal cell carcinoma (BCC) is the most common skin cancer, and the immune system plays a prominent role in tumor appearance and progression. OBJECTIVE To address associations among recent severe life events, childhood parental emotional maltreatment, depression, and messenger RNA (mRNA) coding for immune markers associated with BCC tumor progression and regression. DESIGN We collected information about early parent-child experiences, severe life events in the past year as assessed by the Life Events and Difficulties Schedule, depression, and mRNA for immune markers associated with BCC tumor progression and regression from patients with BCC tumors. SETTING University medical center. PARTICIPANTS Ninety-one patients with BCC (ages, 23-92 years) who had a previous BCC tumor. MAIN OUTCOME MEASURES The expression of 4 BCC tumor mRNA markers (CD25, CD3ε, intercellular adhesion molecule 1, and CD68) that have been linked to BCC tumor progression and regression were assessed in BCC tumor biopsy specimens. RESULTS Both maternal and paternal emotional maltreatment interacted with the occurrence of severe life events to predict the local immune response to the tumor (adjusted P = .009 and P = .03, respectively). Among BCC patients who had experienced a severe life event within the past year, those who were emotionally maltreated by their mothers (P = .007) or fathers (P = .02) as children had a poorer immune response to the BCC tumor. Emotional maltreatment was unrelated to BCC immune responses among those who did not experience a severe life event. Depressive symptoms were not associated with the local tumor immune response. CONCLUSIONS Troubled early parent-child relationships, in combination with a severe life event in the past year, predicted immune responses to a BCC tumor. The immunoreactivity observed in BCCs and the surrounding stroma reflects an anti-tumor-specific immune response that can be altered by stress.

Stress-associated immunomodulation and its implications for responses to vaccination.

The rapidly growing field of psychoneuroimmunology involves the elucidation of the complex interactions between the CNS, the endocrine system and the immune system and its effects on health. Although the mechanisms involved in this bidirectional communication is not yet fully understood, studies in psychoneuroimmunology have shown that stress, through the hypothalamic–pituitary–adrenal and the sympathetic–adrenal–medullary axes, can induce modulation of the immune system. In this review, we discuss human studies and animal models, which focus on psychological stress and its effects on the immune response to vaccination, emphasizing the implications of these effects on health.

Multiple pathways for Epstein-Barr virus episome loss from nasopharyngeal carcinoma

Epstein‐Barr virus (EBV) is the prototypical example for episomal persistence of genetic information. Yet, little is known about how this viral episome is lost. Episome loss occurs naturally in nasopharyngeal carcinoma (NPC) upon explantation into culture. Using whole‐genome profiling, we found evidence for 2 different pathways of episome loss: (i) rapid loss of the entire episome or (ii) successive mutation/deletion of the episome until at least 1 essential cis‐element is destroyed. This second phenotype was seen in a clone of HONE‐1 NPC cells that maintains the EBV episome for prolonged time in culture. The conceptual insights provided by our quantitative analysis should aid our understanding of mammalian episomes, as well as lead to designs to cure latent viral infection.