Gene expression profiling in human insulinoma tissue: genes involved in the insulin secretion pathway and cloning of novel full-length cDNAs.

2004 ◽  
Vol 11 (2) ◽  
pp. 295-303 ◽  
Author(s):  
X-C Wang ◽  
S-Y Xu ◽  
X-Y Wu ◽  
H-D Song ◽  
Y-F Mao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Insulin Secretion ◽  
Molecular Biology ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Cdna Array ◽  
Full Length ◽  
Secretion Pathway ◽  
Est Sequencing ◽  
Human Insulinoma

Insulinoma is a clinically common cause of organic hypoglycemia. The prominent characteristic of insulinoma is endogenous hyperinsulinism. Until now, the molecular biology of human insulinoma has been little understood. In this study, gene expression profiling of human insulinoma was established by expressed sequence tag (EST) sequencing and cDNA array. A total of 2063 clones were obtained, of these, 1589 clones were derived from EST sequencing, 975 clones were derived from cDNA array and 501 clones were shared by the two methods. G protein alpha-stimulating activity polypeptide (Gsalpha) and carboxypeptidase E (CPE) were the most highly expressed genes in human insulinoma, as derived by EST sequencing and cDNA array respectively. The genes involved in the protein/insulin secretion pathway were strongly expressed in human insulinoma tissue. Meanwhile, eight full-length cDNAs of novel genes were cloned and sequenced. The results demonstrated the molecular biology of human insulinoma tissue at the level of transcript abundance and validated the efficacy of EST sequencing combined with cDNA array in the construction of gene expression profiling. In conclusion, the predominance of the genes participating in the secretory pathway suggested that regulation of secretion might be a major mechanism by which insulin release is abnormally increased in patients with insulinomas. It was also concluded that overexpression of the Gsalpha gene played an important role in the pathogenesis of insulinoma.

scholarly journals 256CONSTRUCTION OF A BOVINE CDNA ARRAY TO MONITOR GENE EXPRESSION PROFILES IN BOVINE PREIMPLANTATION EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 248
Author(s):  
C. Wrenzycki ◽  
T. Brambrink ◽  
D. Herrmann ◽  
J.W. Carnwath ◽  
H. Niemann
Keyword(s):  
Gene Expression ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Expression Profiles ◽  
Cdna Array ◽  
Preimplantation Embryos ◽  
Average Insert Size ◽  
Rt Pcr ◽  
Public Data

Array technology is a widely used tool for gene expression profiling, providing the possibility to monitor expression levels of an unlimited number of genes in various biological systems including preimplantation embryos. The objective of the present study was to develop and validate a bovine cDNA array and to compare expression profiles of embryos derived from different origins. A bovine blastocyst cDNA library was generated. Poly(A+)RNA was extracted from in vitro-produced embryos using a Dynabead mRNA purification kit. First-strand synthesis was performed with SacIT21 primer followed by randomly primed second-strand synthesis with a DOP primer mix (Roche) and a global PCR with 35 cycles using SacIT21 and DOP primers. Complementary DNA fragments from 300 to 1500bp were extracted from the gel and normalized via reassoziation and hydroxyapatite chromatography. Resulting cDNAs were digested with SacI and XhoI, ligated into a pBKs vector, and transfected into competent bacteria (Stratagene). After blue/white colony selection, plasmids were extracted and the inserts were subjected to PCR using vector specific primers. Average insert size was determined by size idenfication on agarose gels stained with ethidium bromide. After purification via precipitation and denaturation, 192 cDNA probes were double-spotted onto a nylon membrane and bound to the membrane by UV cross linking. Amplified RNA (aRNA) probes from pools of three or single blastocysts were generated as described recently (Brambrink et al., 2002 BioTechniques, 33, 3–9) and hybridized to the membranes. Expression profiles of in vitro-produced blastocysts cultured in either SOF plus BSA or TCM plus serum were compared with those of diploid parthenogenetic ones generated by chemical activation. Thirty-three probes have been sequenced and, after comparison with public data bases, 26 were identified as cDNAs or genes. Twelve out of 192 (6%) seem to be differentially expressed within the three groups;; 7/12 (58%) were down-regulated, 3/12 (25%) were up-regulated in SOF-derived embryos, and 2/12 (20%) were up-regulated in parthenogenetic blastocysts compared to their in vitro-generated counterparts. Three of these genes involved in calcium signaling (calmodulin, calreticulin) and regulation of actin cytoskeleton (destrin) were validated by semi-quantitative RT-PCR (Wrenzycki et al., 2001 Biol. Reprod. 65, 309–317) employing poly(A+) RNA from a single blastocyst as starting material. No differences were detected in the relative abundance of the analysed gene transcripts within the different groups. These findings were confirmed employing the aRNA used for hybridization in RT-PCR and showed a good representativity of the selected transcripts. Results indicate that it is possible to construct a homologous cDNA array which could be used for gene expression profiling of bovine preimplantation embryos. Supported by the Deutsche Forschungsgemeinschaft (DFG Ni 256/18-1).

scholarly journals Gene expression profiling of human stenotic aorto-coronary bypass grafts by cDNA array analysis

2003 ◽  
Vol 23 (4) ◽  
pp. 620-625 ◽  
Author(s):  
Michael Hilker ◽  
Tina Längin ◽  
Ulrich Hake ◽  
Franz-Xaver Schmid ◽  
Wlodzimierz Kuroczynski ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Cdna Array ◽  
Coronary Bypass ◽  
Bypass Grafts ◽  
Array Analysis ◽  
Coronary Bypass Grafts

Gene expression profiling of malignant mesothelioma cell lines: cDNA array study

2001 ◽  
Vol 91 (4) ◽  
pp. 492-496 ◽  
Author(s):  
Eeva Kettunen ◽  
Anna-Maria Niss�n ◽  
Tiina Ollikainen ◽  
Matti Taavitsainen ◽  
Johanna Tapper ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profiling ◽  
Cell Lines ◽  
Malignant Mesothelioma ◽  
Expression Profiling ◽  
Cdna Array ◽  
Mesothelioma Cell

The gene expression profiling of sporadic pheochromocytoma and novel full-length cDNAs cloning.

2003 ◽  
pp. 621-627
Author(s):  
Y-S Yang ◽  
H-D Song ◽  
Y-D Peng ◽  
Q-H Huang ◽  
R-Y Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adrenal Gland ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Molecular Mechanisms ◽  
Sequence Data ◽  
Full Length ◽  
Familial Pheochromocytoma ◽  
Sporadic Pheochromocytoma

Pheochromocytoma is a chromaffin cell neoplasm that typically causes symptoms and signs of episodic catecholamine release. Pheochromocytoma can be divided into two types: familial and sporadic. The molecular mechanisms involved in familial pheochromocytoma have been unraveled, but the detailed molecular mechanism of sporadic pheochromocytoma remains unknown. The present study thus aimed at characterization of gene expression profiling of sporadic pheochromocytoma using expressed sequence tags (ESTs), and established a preliminary catalog of genes expressed in the tumor. In total, 4115 ESTs were generated from the tumor library. The gene expression profilings of the pheochromocytoma and the normal adrenal gland were compared, and 341 genes were identified to be significantly expressed differently between the two libraries. Interestingly, 16 known genes participating in cell division or apoptosis were notably differently expressed between the tumor and the normal adrenal gland. Twenty-four novel full-length cDNAs were cloned from the tumor library and five of them were significantly up-regulated in the tumor. Some of them may be involved in the tumorigenesis of pheochromocytoma. The sequence data of ESTs and novel full-length cDNAs described in this paper have been submitted to the GeneBank library.

Gene expression profiling of phenylbutyrate induced differentiation of glioma cells by cDNA array

2003 ◽  
Vol 15 (1) ◽  
pp. 38-42
Author(s):  
Li-jun Sun ◽  
Qiang Huang ◽  
Qing Lan ◽  
Zi-wei Du ◽  
Geng-xi Hu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Cdna Array ◽  
Glioma Cells ◽  
Induced Differentiation

Gene expression profiling of ganglioglioma malignant progression by cDNA array

2005 ◽  
Vol 17 (1) ◽  
pp. 11-16
Author(s):  
Quan-bin Zhang ◽  
Qiang Huang ◽  
Jun Dong ◽  
Ai-dong Wang ◽  
Ji-yong Sun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Cdna Array ◽  
Malignant Progression
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