scholarly journals Microarray and Suppression Subtractive Hybridization Analyses of Gene Expression in Pheochromocytoma Cells Reveal Pleiotropic Effects of Pituitary Adenylate Cyclase-Activating Polypeptide on Cell Proliferation, Survival, and Adhesion

Endocrinology ◽  
2003 ◽  
Vol 144 (6) ◽  
pp. 2368-2379 ◽  
Author(s):  
Luca Grumolato ◽  
Abdel G. Elkahloun ◽  
Hafida Ghzili ◽  
David Alexandre ◽  
Cédric Coulouarn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Cell Differentiation ◽  
Adenylate Cyclase ◽  
Pc12 Cells ◽  
Suppression Subtractive Hybridization ◽  
Global Gene Expression ◽  
Subtractive Hybridization ◽  
Neuroendocrine Cells ◽  
Pituitary Adenylate Cyclase

Abstract Pituitary adenylate cyclase-activating polypeptide (PACAP) exerts trophic effects on several neuronal, neuroendocrine, and endocrine cells. To gain insight into the pattern of the transcriptional modifications induced by PACAP during cell differentiation, we studied the effects of this neuropeptide on rat pheochromocytoma PC12 cells. We first analyzed the transcriptome of PC12 cells in comparison to that of terminally differentiated rat adrenomedullary chromaffin cells, using a high-density microarray, to identify genes associated with the proliferative phenotype that are possible targets of PACAP during differentiation of sympathoadrenal normal and tumoral cells. We then studied global gene expression in PC12 cells after 48 h of exposure to PACAP, using both cDNA microarray and suppression subtractive hybridization technologies. These complementary approaches resulted in the identification of 75 up-regulated and 70 down-regulated genes in PACAP-treated PC12 cells. Among the genes whose expression is modified in differentiated cells, a vast majority are involved in cell proliferation, survival, and adhesion/motility. Expression changes of most of these genes have been associated with progression of several neoplasms. A kinetic study of the effects of PACAP on some of the identified genes showed that the neuropeptide likely exerts early as well as late actions to achieve the gene expression program necessary for cell differentiation. In conclusion, the results of the present study underscore the pleiotropic role of PACAP in cell differentiation and provide important information on novel targets that could mediate the effects of this neuropeptide in normal and tumoral neuroendocrine cells.

Costimulatory Blockade (CSB) during Mixed Lymphocyte Reaction (MLR) Prevents IL27 Upregulation and Signalling.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4878-4878
Author(s):  
Gullu Gorgun ◽  
Patrick Philpot ◽  
Kamila Naxerova ◽  
Isaac Kohane ◽  
Lee Nadler ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
T Cells ◽  
Cell Differentiation ◽  
Differentially Expressed Genes ◽  
Ex Vivo ◽  
Global Gene Expression ◽  
Th1 Cell ◽  
Cellular Pathways

Abstract Numerous methods for selectively depleting or manipulating alloantigen (alloAg) specific CD4 T cells (CD4 T) are being studied for their potential in improving transplant outcomes by limiting GVHD or graft rejection. Although some methods target specific molecules (e.g. costimulatory receptors and ligands, activation antigens), the effects of these methods on “off-target” CD4 T cellular pathways and functions as well as effects on other PBMC, direct or indirect, have not been well characterized. Such effects may impact significantly on clinical efficacy and/or may shed light on the mechanism of action of manipulated PBMC in a complex in vivo milieu. To understand the molecular impact on bystander PBMC of inducing alloAg specific CD4 T anergy by CSB via disruption of the positive costimulatory signal between B7.1 and B7.2 on antigen presenting cells (APC) and CD28 on CD4 T by humanized anti B7.1/B7.2 monoclonal antibodies (MoAbs), we have used global gene expression profiling (GEP). Mimicking our ex vivo clinical anergization protocol, we used PBMC obtained from fully-HLA mismatched healthy volunteers (n=12 pairs) to perform an ex vivo primary MLR +/− anti B7.1/B7.2 MoAbs. CD28:B7 CSB inhibited mean proliferation by 73% after 72 hours of MLR. GEP was performed using Affymetrix hu133 plus2 chips on monocytes (Mo), CD4 and CD8 T cells, and B and NK cells purified from the MLR. Differentially expressed genes between cells from MLR +/− CSB were determined by SAM and EBAM analysis. Despite the low published frequency (1–10%) of alloAg specific CD4 T, we could detect global gene expression variance between CD4 T isolated from MLR +/− CSB (P≤0.05) suggesting effects on non-alloAg specific CD4 T. Use of the Ingenuity pathway analyzer to classify these differentially expressed genes by specific cellular pathways showed most were involved in cell proliferation and differentiation. Differences in IL27 downstream signaling molecules (DSM) in Mo and CD4 T were pronounced. IL27 is a member of IL12 cytokine family produced by APC and is a heterodimer of p28 and EBV-Induced gene 3 (EBI3). The IL27 receptor (IL27R) WSX1 is expressed on CD4 T. IL27 regulates adaptive immunity by controlling T cell proliferation, Th1 cell differentiation and IFNγ synthesis. Both p28 and EBI3 transcriptional and translational levels were decreased in Mo from MLR + CSB. Expression of STAT3, an IL27 pathway DSM was also decreased. After CD28:B7 CSB, CD4 T exhibited decreased expression of the IL27R and also inactivation of IL27 DSM including pSTAT1 and NFκB at both the transcriptional and translational levels. Consistent with a negative effect on Th1 differentiation, intracytoplasmic cytokine (ICC) analysis showed decreased expression of type 1 cytokines IL2 and IFNγ in CD4 T from MLR + CSB. ICC also showed decreased expression of the type 1 cytokine IL15 and increased expression of the type 2 cytokine IL10 in Mo from MLR + CSB. These results suggest that CD28:B7 signaling during MLR is required for Mo production of IL27. Decreased expression of IL27 and its DSMs pSTAT1 and NFκB1 after CSB with antiB7 MoAbs may contribute to CD4 T alloAg anergy induction by suppressing effector cytokines and Th1 cell differentiation. The observation that CD28:B7 modulates IL27 and IL27R emphasizes that targeted therapies used in the complex environment of human PBMC may have effects not predicted by in vitro clonal systems. Such effects may be important in the functional outcome of the intervention.

Changes of dopamine content and cell proliferation by dexamethsone via pituitary adenylate cyclase-activating polypeptide in PC12 cell

2007 ◽  
Vol 426 (1) ◽  
pp. 45-48 ◽  
Author(s):  
Ting-Ting Yang ◽  
Chiung-Wen Tsao ◽  
Jin-Shiou Li ◽  
Hung-Tsung Wu ◽  
Chao-Tien Hsu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Adenylate Cyclase ◽  
Pc12 Cell ◽  
Dopamine Content ◽  
Pituitary Adenylate Cyclase

scholarly journals Effects of low-to-moderate ethanol consumption on colonic growth and gene expression in young adult and middle-aged male rats

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243499
Author(s):  
Nicole Wells ◽  
Jacqueline Quigley ◽  
Jeremy Pascua ◽  
Natalie Pinkowski ◽  
Lama Almaiman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Colorectal Cancer ◽  
Cell Proliferation ◽  
Cell Differentiation ◽  
Young Adult ◽  
Cell Growth ◽  
Alcohol Consumption ◽  
Ethanol Consumption ◽  
Ethanol Treatment ◽  
Middle Aged

Excessive alcohol consumption is a risk factor associated with colorectal cancer; however, some epidemiological studies have reported that moderate alcohol consumption may not contribute additional risk or may provide a protective effect reducing colorectal cancer risk. Prior research highlights the importance of proliferation, differentiation, and apoptosis as parameters to consider when evaluating colonic cell growth and tumorigenesis. The present study investigated whether chronic low-to-moderate ethanol consumption altered these parameters of colonic cell growth and expression of related genes. Twenty-four nondeprived young adult (109 days old) and 24 nondeprived middle-aged (420 days old) Wistar rats were randomly assigned to an ethanol-exposed or a water control group (n = 12/group). The ethanol group was provided voluntary access to a 20% v/v ethanol solution on alternate days for 13 weeks. Colon tissues were collected for quantitative immunohistochemical analyses of cell proliferation, differentiation and apoptosis using Ki-67, goblet cell and TUNEL, respectively. Gene expression of cyclin D1 (Ccnd1), Cdk2, Cdk4, p21waf1/cip1 (Cdkn1a), E-cadherin (Cdh1) and p53 were determined by quantitative real-time polymerase chain reaction in colonic scraped mucosa. Ethanol treatment resulted in a lower cell proliferation index and proliferative zone, and lower Cdk2 expression in both age groups, as well as trends toward lower Ccnd1 and higher Cdkn1a expression. Cell differentiation was modestly but significantly reduced by ethanol treatment only in older animals. Overall, older rats showed decreases in apoptosis and gene expression of Cdk4, Cdh1, and p53 compared to younger rats, but there was no observed effect of ethanol exposure on these measures. These findings suggest that low-to-moderate ethanol consumption improves at least one notable parameter in colonic tumorigenesis (cell proliferation) and associated gene expression regardless of age, however, selectively decreased cell differentiation among older subjects.

scholarly journals Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) and Nerve Growth Factor (NGF) Induce PACAP Expression in PC12 Cells

1998 ◽  
Vol 76 ◽  
pp. 171
Author(s):  
Nami Hagihara ◽  
Hitoshi Hashimoto ◽  
Kyohei Yamamoto ◽  
Takashi Fujita ◽  
Norihito Shintani ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Adenylate Cyclase ◽  
Pc12 Cells ◽  
Nerve Growth ◽  
Pituitary Adenylate Cyclase
Sign in / Sign up

Export Citation Format

Share Document