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HOXB7 mRNA is overexpressed in pancreatic ductal adenocarcinomas and its knockdown induces cell cycle arrest and apoptosis.

ABSTRACT: In order to determine the role of HOXB7 gene in cell lines derived from pancreatic ductal adenocarcinoma (MIA PaCa-2 e Capan-1), we performed its silencing utilizing the technique of RNA interference (RNAi). Total RNA derived from the inhibition as well as from parental cells was quantified in Bioanalyzer (Agilent,Santa Clara, CA, USA) and employed in the Agilent platform (4x44K). Proceeded with the guidelines of the One Color Microarray-Based Gene Expression Protocol (Agilent) and with the use of the Agilent Low Input Quick Amp Labeling Kit. HOXB7 knockdown elicited the modulation of several biological processes, especially in the MIA Paca-2 cell line, such as proteasomal ubiquitin-dependent catabolic process, synthesis of amines and cell cycle. Moreover, it showed induction of apoptosis and reduction in cell proliferation by flow cytometry and MTT assay, respectively. In this context, HOXB7 represents another component associated with the extensive network of molecules involved in the tumorigenesis of pancreatic cancer and could be a promising target for future biologic therapies.

ORGANISM(S): Homo sapiens

PROVIDER: GSE46393 | GEO | 2013/07/26

SECONDARY ACCESSION(S): PRJNA199759

REPOSITORIES: GEO

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HOXB7 mRNA is overexpressed in pancreatic ductal adenocarcinomas and its knockdown induces cell cycle arrest and apoptosis.

Project description:In order to determine the role of HOXB7 gene in cell lines derived from pancreatic ductal adenocarcinoma (MIA PaCa-2 e Capan-1), we performed its silencing utilizing the technique of RNA interference (RNAi). Total RNA derived from the inhibition as well as from parental cells was quantified in Bioanalyzer (Agilent,Santa Clara, CA, USA) and employed in the Agilent platform (4x44K). Proceeded with the guidelines of the One Color Microarray-Based Gene Expression Protocol (Agilent) and with the use of the Agilent Low Input Quick Amp Labeling Kit. HOXB7 knockdown elicited the modulation of several biological processes, especially in the MIA Paca-2 cell line, such as proteasomal ubiquitin-dependent catabolic process, synthesis of amines and cell cycle. Moreover, it showed induction of apoptosis and reduction in cell proliferation by flow cytometry and MTT assay, respectively. In this context, HOXB7 represents another component associated with the extensive network of molecules involved in the tumorigenesis of pancreatic cancer and could be a promising target for future biologic therapies. The procedure was performed in duplicate for both cell lines, which were sorted into treated and untreated with RNAi.

2013-07-26 | E-GEOD-46393 | biostudies-arrayexpress

Yarrow and marigold extracts and their effect in pancreatic cancer cells [MIL]

Project description:determine the molecular effect of yarrow and marigold supercritical extracts in gene expression of pancreatic cancer cells (MIA PaCa-2)

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Yarrow and marigold extracts and their effect in pancreatic cancer cells [CAL]

Project description:determine the molecular effect of yarrow and marigold supercritical extracts in gene expression of pancreatic cancer cells (MIA PaCa-2)

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Transcriptome analysis of RNA from cancer cells exposed to chronic hypoxia versus normoxia

Project description:Transcriptome data from six cancer cell lines passaged twenty times in normoxia (21% oxygen) or hypoxia (1% oxygen) Transcriptome analysis was conducted to determine the effect of chronic, long passaging, at 1% oxygen (hypoxia) on gene expression and alternative splicing in six cancer cell lines. The cancer cell lines included were two pancreatic (MIA PaCa-2, Capan-1), one lung (H226), one breast (MCF-7), one colon (HT-29), and one ovarian (SKOV-3). They were selected from a panel of 12 cancer cell lines since they showed the highest and lowest sensitivity to hypoxia based on their expression levels of a hypoxia 8-gene signature.

2023-03-29 | GSE222048 | GEO

MIA PaCa-2 cells treated with Gemcitabine

Project description:Pancreatic cancer cells (MIA PaCa-2) were treated with Gemcitabine and overall protein changes were studied

2022-09-15 | PXD030861 | Pride

Identification of target genes of miR-193b

Project description:Searching of target genes of miR-193b by transcriptome assay using 44K Whole Human Genome Microarray system (Agilent Technologies, Palo Alto, CA) resulted in finding of several candidate genes. To search of targets genes of miR-193b, we performed transcriptome analysis to compare expression profiles between human pancreatic cancer cells, MIA PaCa-2, transfected with precursor of miR-193b and those transfected with a negative control precursor.

2012-03-14 | E-GEOD-25215 | biostudies-arrayexpress

CRISPR deletion of ELAVL1 (HuR) in MIA PaCa-2 in vitro cells and in vivo tumors

Project description:To investigate the impact of the mRNA stabilty factor HuR on the pancreatic cancer transcriptome in MIA PaCa-2 in vitro cell line and in vivo tumors at multiple time points of tumor growth

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Shotgun proteomic analysis of pancreatic cancer cell lines vs. normal epithelial ductal pancreatic cells

Project description:We have run a shotgun proteomic analysis of cell lysates from pancreatic cancer cell lines (PANC-1, PaCa-44, MIA PaCa-2 and BxPC-3) vs. normal epithelial ductal pancreatic cells (HPDE) in LC-MS/MS. No labelling was performed and digestion was done with Trypsin/Lys-C mix endoproteinase.

2020-02-07 | MSV000084935 | MassIVE

MicroRNA screening in human pancreatic carcinoma cell lines

Project description:No reliable predictors of susceptibility to gemcitabine chemotherapy exist in pancreatic ductal adenocarcinoma. MicroRNAs (miR) are epigenetic gene regulators with tumorsuppressive or oncogenic roles in various carcinomas. This study assesses chemoresistant PDAC for its specific miR expression pattern. Gemcitabine-resistant variants of two mutant p53 human pancreatic adenocarcinoma cell lines were established. MicroRNA screening was investigated by microarray. Gemcitabine-resistant PANC-1 (PANC-1-GR) and MIA-PaCa-2 (MIA-PaCa-2-GR) cell clones were produced by exposing the parental cells to repeated pulsatile gemcitabine treatment over 3 days with constant sublethal concentrations followed by recovery-periods with agent-free medium until the cells recovered exponentially. Parental PANC-1 cells were treated with 0.4ÂµM gemcitabine cycles for approximately 9 months. Parental MIA-PaCa-2 cells were exposed to 0.06ÂµM gemcitabine cycles for approximately 12 months. Affymetrix GeneChip miRNA microarrays (Affymetrix UK Ltd., High Wycombe, UK) were performed in parental and chemoresistant PANC-1 and MIA-PaCa-2 cells after 29 chemotherapy cycles using the manufacturersÂ´ protocols. The samples were prepared from 1Âµg of total-RNA in accordance with the Affymetrix FlashTag Biotin HSR RNA Labeling Kit. The targets were hybridized overnight to Affymetrix GeneChip miRNA arrays. Following hybridization, the arrays were washed and stained using the Affymetrix GeneChip Fluidics Station 450 and scanned using the Affymetrix GeneChip Scanner 3000 7G. Microarray data quality was checked as recommended by the manufacturer and by the quality metrics in the Partek Genomics Suite software (Partek Inc., St. Louis, MO).

2015-11-03 | E-GEOD-74565 | biostudies-arrayexpress

Pachymic acid inhibits growth and induces apoptosis of pancreatic cancer in vitro and in vivo by targeting ER stress

Project description:Background: Pachymic acid (PA) is a purified triterpene extracted from medicinal fungus Poria cocos. In this paper, we investigated the anticancer effects of PA on human chemotherapy resistant pancreatic cancer cells. Methods: Gemcitabine-resistant pancreatic cancer cells PANC-1 and MIA PaCa-2 were used, along with a xenograft model of MIA PaCa-2 cells implanted in mice. Apoptosis was assessed by quantitation of cytoplasmic histone-associated DNA fragments and expression of cleaved PARP. Differential expression of genes was identified using comparative DNA microarray analysis. Protein levels were determined by immunoblotting. Toxicology studies in vivo were assessed by detecting pathological changes in organs and liver enzyme profiles in plasma. Tumor tissues were analyzed by quantification of apoptotic bodies, qRT-PCR and immunoblotting. Principal Findings: PA induced endoplasmic reticulum (ER) stress in chemotherapy resistant pancreatic cancer cells through activation of heat shock response and unfolded protein response related genes, which further triggered apoptosis. The involvement of ER stress was confirmed by increasing expression of XBP-1s, ATF4, Hsp70, CHOP and phospho-eIF2M-NM-1. Moreover, 25 mg kgM-bM-^HM-^R1 of PA significantly suppressed MIA PaCa-2 tumor growth in vivo without toxicity, which correlated with induction of apoptosis, ER stress related genes and proteins expression. Conclusions: Growth inhibition and induction of apoptosis by PA in chemotherapy resistant pancreatic cancer cells were associated with ER stress activation both in vitro and in vivo. Pancreatic cancer cell line treated with pachymic acid vs. control (untreated)

2014-12-12 | E-GEOD-64111 | biostudies-arrayexpress

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