Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Expression data from mouse insulinoma cells

ABSTRACT: We previously isolated a subclone, MIN6 clone 4, from the parental MIN6 cells, that shows well-regulated insulin secretion in response to glucose, glybenclamide, and KCl, even after prolonged culture. To investigate the molecular mechanisms responsible for preserving GSIS in this subclone, we compared four groups of MIN6 cells: Pr-LP (parental MIN6, low passage number), Pr-HP (parental MIN6, high passage number), C4-LP (MIN6 clone 4, low passage number), and C4-HP (MIN6 clone 4, high passage number). Based on their capacity for GSIS, we designated the Pr-LP, C4-LP, and C4-HP cells as M-bM-^@M-^\responder cells.M-bM-^@M-^] In a DNA microarray analysis, we identified a group of genes with high expression in responder cells (M-bM-^@M-^\responder genesM-bM-^@M-^]), but extremely low expression in the Pr-HP cells. MIN6 clone 4 cells are a subclone isolated from low-passage-number parental MIN6 cells by the limiting dilution method (JM, unpublished). This subclone was maintained in the same culture conditions as the parental cells, and retained good GSIS even after 6 months of continuous culture. For the low-passage-number parental MIN6 cells (Pr-LP), we used cells passaged 17-20 times; for the high-passage-number MIN6 cells (Pr-HP), we used cells passaged 35-40 times. Seventeen to 20 passages were also used for the low-passage-number MIN6 clone 4 cells (C4-LP), and the high-passage ones (C4-HP) were used after 40 to 50 passages.

ORGANISM(S): Mus musculus

SUBMITTER: Eiji Yamato

PROVIDER: E-GEOD-43774 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Microarray analysis of novel candidate genes responsible for glucose-stimulated insulin secretion in mouse pancreatic β cell line MIN6.

Yamato Eiji E Tashiro Fumi F Miyazaki Jun-ichi J

PloS one 20130403 4

Elucidating the regulation of glucose-stimulated insulin secretion (GSIS) in pancreatic islet β cells is important for understanding and treating diabetes. MIN6 cells, a transformed β-cell line derived from a mouse insulinoma, retain GSIS and are a popular in vitro model for insulin secretion. However, in long-term culture, MIN6 cells' GSIS capacity is lost. We previously isolated a subclone, MIN6 clone 4, from the parental MIN6 cells, that shows well-regulated insulin secretion in response to g ...[more]

PMID: 23560115

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Project description:MicroRNAs (miRNAs) are small noncoding RNAs that negatively regulate gene expression post-transcriptionally. They play a critical role in developmental and physiological processes and have been implicated in the pathogenesis of several diseases including cancer. To identify miRNA signatures associated with different stages of neoplastic development, we examined the expression profile of 776 primate miRNAs in the following cells: primary African green monkey kidney (pAGMK) cells; spontaneously immortalized, non-tumorigenic, low-passage VERO cells (10-87 LP); tumorigenic, high-passage VERO cells (10-87 HP); and a cell line (10-87 T) derived from a 10-87 HP cell tumor xenograft in athymic nude mice. When compared with pAGMK cells, the majority of miRNAs were expressed at lower levels in 10-87 LP, 10-87 HP, and 10-87 T cells. We identified 10 up-regulated miRNAs whose level of expression correlated with VERO cell evolution from a non-tumorigenic phenotype to a tumorigenic phenotype. Several miRNAs that were components of the tumorigenic phenotype-specific signatures in our AGMK model are also found in a variety of human tumors. This may prove to be of general relevance to the biology of neoplastic development as it occurs both in vivo as well as in vitro. In addition, one or more of these miRNAs could be potential biomarkers for the expression of the tumorigenic phenotype of VERO cells. The spontaneousely transformed VERO cells, non-tumorigenic, were pasasged at low density in culture up to 250. The high passage (p250) was found to be tumorigenic. The cell line from xenograft of high passage was also established. We then evaluated patterns of miRNA expression in pAGMK cells and in derivatives of the 10-87 VERO cell line (10-87 LP cells, 10-87 HP cells, and 10-87 T cells) in an attempt to identify the miRNAs whose altered expression might correlate with, and perhaps be involved in, the evolution of the neoplastic phenotypes that occurred during passage of these AGMK cells in tissue culture. performed high-throughput miRNA profiling to audit the expression level of miRNAs in pAGMK cells and in VERO cells at non-tumorigenic and tumorigenic stages of neoplastic development. The analysis involved pAGMK cells, non-tumorigenic 10-87 low-passage VERO cells (10-87 LP) tumorigenic, high-passage VERO cells (10-87 HP) and a cell line (10-87 T) derived from a 10-87 HP cell tumor xenograft in athymic nude mice.

Dataset Information

Expression data from mouse insulinoma cells

Publications

Microarray analysis of novel candidate genes responsible for glucose-stimulated insulin secretion in mouse pancreatic β cell line MIN6.

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