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Kieffer N Schmitz M Plançon S Margue C Huselstein F Grignard G Dippel W Nathan M Giacchi S Scheiden R 《International journal of oncology》2005,26(6):1549-1558
A major technical challenge related to gene expression profiling of tissue samples is the difficulty of procuring selected cell populations from tissues that by nature are heterogeneous, such as prostate tissue. In this study we have examined the expression of integrin-linked kinase (ILK) mRNA in prostate adenocarcinoma cells versus normal prostate epithelial cells in order to determine whether ILK could be used as a reference marker gene for prostate adenocarcinoma cell mRNA isolation. Using laser microdissection (LMD) technology and real-time PCR, together with immunohistochemistry, we have analyzed ILK mRNA expression in epithelial cells isolated from frozen prostate biopsy specimens as well as 4 prostate cell lines (RWPE-1, LNCaP, PC-3 and DU 145) and correlated ILK mRNA expression with ILK protein expression. We demonstrate that quantitative upregulation of ILK mRNA expression in prostate epithelial cells derived from prostate tissue correlated with ILK protein expression and with the histopathology diagnosis of prostate adenocarcinoma. We further show that the level of ILK overexpression was directly influenced by the method used to isolate prostate adenocarcinoma cells (bulk tissue versus LMD dissected cells). These data provide evidence that ILK mRNA is quantitatively upregulated in prostate adenocarcinoma cells versus normal epithelial cells and is therefore a useful internal reference gene marker to evaluate the quality of prostate adenocarcinoma cell derived mRNA used for large scale prostate cancer cDNA gene profiling. 相似文献
58.
Walwyn W Maidment NT Sanders M Evans CJ Kieffer BL Hales TG 《Molecular pharmacology》2005,68(6):1688-1698
It is not clear whether primary afferent neurons express functional cell-surface opioid receptors. We examined delta receptor coupling to Ca2+ channels in mouse dorsal root ganglion neurons under basal conditions and after receptor up-regulation. [D-Ala2,Phe4,Gly5-ol]-enkephalin (DAMGO), [D-Ala2,D-Leu5]-enkephalin (DADLE), trans-(+/-)-3,4-dichloro-N-methyl-N-(2-[1-pyrrolidinyl]cyclohexyl) benzene-acetamide methanesulfonate (U-50,488H; 1 microM), and baclofen (50 microM) inhibited Ca2+ currents, whereas the -selective ligands [D-Pen2,Pen5]-enkephalin (DPDPE) and deltorphin II (1 microM) did not. The effect of DADLE (1 microM) was blocked by the mu-antagonist D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP; 300 nM) but not by the -antagonist Tyr-1,2,3,4-tetrahydroisoquinoline-Phe-Phe-OH (300 nM), implicating mu receptors. Despite a lack of functional delta receptors, flow cytometry revealed cell-surface receptors. We used this approach to identify conditions that up-regulate receptors, including mu receptor gene deletion in dorsal root ganglion neurons of mu-/- mice and 18-h incubation of mu+/+ neurons with CTAP followed by brief (10-min) DPDPE exposure. Under these conditions, the expression of cell-surface delta receptors was up-regulated to 149 +/- 9 and 139 +/- 5%, respectively; furthermore, DPDPE and deltorphin II (1 microM) inhibited Ca2+ currents in both cases. Viral replacement of mu receptors in mu-/- neurons reduced delta receptor expression to mu+/+ levels, restored the inhibition of Ca2+ currents by DAMGO, and abolished receptor coupling. Our observations suggest that receptor-Ca2+ channel coupling in primary afferent fibers may have little functional significance under basal conditions in which mu receptors predominate. However, up-regulation of cell-surface delta receptors induces their coupling to Ca2+ channels. Pharmacological approaches that increase functional delta receptor expression may reveal a novel target for analgesic therapy. 相似文献
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Racial and Ethnic Approaches to Community Health (REACH) Detroit partnership: improving diabetes-related outcomes among African American and Latino adults 下载免费PDF全文
Two Feathers J Kieffer EC Palmisano G Anderson M Sinco B Janz N Heisler M Spencer M Guzman R Thompson J Wisdom K James SA 《American journal of public health》2005,95(9):1552-1560
OBJECTIVES: We sought to determine the effects of a community-based, culturally tailored diabetes lifestyle intervention on risk factors for diabetes complications among African Americans and Latinos with type 2 diabetes. METHODS: One hundred fifty-one African American and Latino adults with diabetes were recruited from 3 health care systems in Detroit, Michigan, to participate in the Racial and Ethnic Approaches to Community Health (REACH) Detroit Partnership diabetes lifestyle intervention. The curriculum, delivered by trained community residents, was aimed at improving dietary, physical activity, and diabetes self-care behaviors. Baseline and postintervention levels of diabetes-specific quality-of-life, diet, physical activity, self-care knowledge and behaviors, and hemoglobin A1C were assessed. RESULTS: There were statistically significant improvements in postintervention dietary knowledge and behaviors and physical activity knowledge. A statistically significant improvement in A1C level was achieved among REACH Detroit program participants (P<.0001) compared with a group of patients with diabetes in the same health care system in which no significant changes were observed (P=.160). CONCLUSIONS: A culturally tailored diabetes lifestyle intervention delivered by trained community residents produced significant improvement in dietary and diabetes self-care related knowledge and behaviors as well as important metabolic improvements. 相似文献
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Tabarin A Diz-Chaves Y Chaves YD Carmona Mdel C Catargi B Zorrilla EP Roberts AJ Coscina DV Rousset S Redonnet A Parker GC Inoue K Ricquier D Pénicaud L Kieffer BL Koob GF 《Diabetes》2005,54(12):3510-3516
Using pharmacological tools, a role for opioid receptors in the regulation of food intake has been documented. However, the involvement of specific receptor subtypes remains questionable, and little information is available regarding a role for opioid receptors in energy metabolism. Using adult male mice lacking the mu-opioid receptor (MOR) gene (MOR-/-), we show that the MOR is not essential for the maintenance of normal levels of ad libitum food intake but does modulate the efficiency of energy storage during high-fat diets through the regulation of energy partitioning. When fed a regular diet, MOR-/- mice displayed only subtle alterations in energy homeostasis, suggesting a relative overuse of fat as a fuel source in the fed state. When fed a high-fat diet, MOR-/- mice were resistant to obesity and impaired glucose tolerance, despite having similar energy intake to wild-type mice. This resistance to obesity was associated with a strong induction of the expression of key mitochondrial enzymes involved in fatty acid oxidation within skeletal muscle. This metabolic role of the MOR, which is consistent with the properties of a "thrifty gene," suggests that the MOR pathway is a potential target for pharmacological intervention in the treatment of obesity associated with the intake of fatty diets. 相似文献