Increased Expression of Cyclin-Dependent Kinase-Interacting Protein p21 during Tamoxifen-Induced Hepatocarcinogenesis in Female Rats

Kasahara, Toshihiko; Kakinuma, Chihaya; Kuwayama, Chitose; Hashiba, Masamichi; Harada, Tsuyoshi; Degawa, Masakuni

doi:10.1248/jhs.51.185

Cited by 4 publications

(2 citation statements)

References 18 publications

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“…For normalization in the 2 −ΔΔ C t method the hypoxanthine guanine phosphoribosyl transferase (HPRT1) gene was employed. The mRNA expression level of selected genes was determined by the following primer pairs (forward/reverse) at the respective annealing temperature given in brackets: HPRT1 TGACACTGGCAAAACAATGCA/GGTCCTTTTCACCAGCAAGCT (62 °C), [ 24 ]; SOD2 TGGCTTGGTTTCAATAAGGAA/AGCGTGCTCCCACACATCAAT (58 °C), (RTPrimerDB); p21 CCTGTCACTGTCTTGTACCCT/GCGTTTGGAGTGGTAGAAATCT (52 °C), [ 25 ]; IFNβ1 GCCGCATTGACCATCTAT/GTCTCATTCCAGCCAGTG (60 °C); IFNλ1 GCAGGTTCAAATCTCTGTCACC/AAGACAGGAGAGCTGCAACTC (60 °C); IFNλ2/3 GCCACATAGCCCAGTTCAAG/TGGGAGAGGATATGGTGCAG (60 °C); viperin GAGAGCCATTTCTTCAAGACC/CTATAATCCCTACACCACCTCC (60 °C); IFIT1 AAAAGCCCACATTTGAGGTG/GAAATTCCTGAAACCGACCA (60 °C); IFITM1 CCAAGGTCCACCGTGATTAAC/ACCAGTTCAAGAAGAGGGTGTT (56 °C), [ 26 ]; IFITM3 GATGTGGATCACGGTGGAC/AGATGCTCAAGGAGGAGCAC (55 °C); ISG15 CTGTTCTGGCTGACCTTCG/GGCTTGAGGCCGTACTCC (56 °C), [ 26 ]; …”

Section: Methodsmentioning

confidence: 99%

Rubella Virus Strain-Associated Differences in the Induction of Oxidative Stress Are Independent of Their Interferon Activation

Zobel

Lorenz

Frascaroli

et al. 2018

Viruses

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Rubella virus (RV) infection impacts cellular metabolic activity in a complex manner with strain-specific nutritional requirements. Here we addressed whether this differential metabolic influence was associated with differences in oxidative stress induction and subsequently with innate immune response activation. The low passaged clinical isolates of RV examined in this study induced oxidative stress as validated through generation of the reactive oxygen species (ROS) cytoplasmic hydrogen peroxide and mitochondrial superoxide. The addition of the cytoplasmic and mitochondrial ROS scavengers N-acetyl-l-cysteine and MitoTEMPO, respectively, reduced RV-associated cytopathogenicity and caspase activation. While the degree of oxidative stress induction varied among RV clinical isolates, the level of innate immune response and interferon-stimulated gene activation was comparable. The type III IFNs were highly upregulated in all cell culture systems tested. However, only pre-stimulation with IFN β slightly reduced RV replication indicating that RV appears to have evolved the ability to counteract innate immune response mechanisms. Through the data presented, we showed that the ability of RV to induce oxidative stress was independent of its capacity to stimulate and counteract the intrinsic innate immune response.

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Section: Methodsmentioning

confidence: 99%

Rubella Virus Strain-Associated Differences in the Induction of Oxidative Stress Are Independent of Their Interferon Activation

Zobel

Lorenz

Frascaroli

et al. 2018

Viruses

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“…Copies of β-actin mRNA were quantified and presented as copy number per total RNA (μg). Standard DNA and primer sets for β-actin were prepared according to Kasahara et al (2005).…”

Section: Preparation Of Cdna and Real-time Pcrmentioning

confidence: 99%

Evaluation of Methods for Duration of Preservation of Rna Quality in Rat Liver Used for Transcriptome Analysis

et al. 2006

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-In The Toxicogenomics Project, about 150 chemicals are administered to rats, and gene expression in the liver analyzed by Affymetrix GeneChip and stored in the database. As the quality of RNA greatly influences the accuracy of gene expression data, conditions of the storage of the sample are very important. Recently, an RNA stabilization solution, RNAlater ® , has become commercially available. In this study, the new storage method was compared with the traditional storage method (stored in freezer or liquid nitrogen) under various conditions by looking at the degradation of RNA assessed by its total yield, OD260/280 ratio, 28S/18S ratio, and quantity of β-actin. It was confirmed that RNAlater ® preserved the liver tissue sample by maintaining the quality of RNA for one year (in liquid N 2 or −80°C), for 3 days (4°C), or for 2 hr (room temperature) without degradation of RNA. Quality of RNA samples dissolved in buffer RLT and stored at −20°C tended to decrease, but samples stored at −80°C were almost equivalent to those stored in liquid nitrogen. In conclusion, we recommend the following procedure for preservation of liver tissue for extraction of RNA: 1) tissues removed should be put into chilled RNAlater ® as soon as possible; 2) samples in RNAlater ® must be stored overnight or longer at 4°C and can be left for as long as 2 weeks without freezing; 3) samples in RNAlater ® can be stored for at least one year under less than −20°C and 4) samples dissolved in buffer RLT can be preserved at least for one year under −80°C.

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Validation and application of normalization factors for gene expression studies in rubella virus‐infected cell lines with quantitative real‐time PCR

Chey¹,

Claus²,

Liebert³

2010

J of Cellular Biochemistry

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Reference genes are generally employed in real-time quantitative PCR (RT-qPCR) experiments to normalize variability between different samples. The aim of this study was to identify and validate appropriate reference genes as internal controls for RT-qPCR experiments in rubella virus (RV)-infected Vero and MCF-7 cell lines using SYBR green fluorescence. The software programs geNorm and NormFinder and the DeltaDeltaC(t) calculation were used to determine the expression stability and thus reliability of nine suitable reference genes. HPRT1 and HUEL, and HUEL and TBP were identified to be most suitable for RT-qPCR analysis of RV-infected Vero and MCF-7 cells, respectively. These genes were used as normalizers for transcriptional activity of selected cellular genes. The results confirm previously published microarray and Northern blot data, particularly on the transcriptional activity of the cyclin-dependent kinase inhibitor p21 and the nuclear body protein SP100. Furthermore, the mRNA level of the mitochondrial protein p32 is increased in RV-infected cells. The effect on cellular gene transcription by RV-infection seems to be cell line-specific, but genes of central importance for viral life cycle appear to be altered to a similar degree. This study does not only provide an accurate and flexible tool for the quantitative analysis of gene expression patterns in RV-infected cell lines. It also indicates, that the suitability of a reference gene as normalizer of RT-qPCR data and the host-cell response to RV-infection are strictly cell-line specific.

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Increased Expression of Cyclin-Dependent Kinase-Interacting Protein p21 during Tamoxifen-Induced Hepatocarcinogenesis in Female Rats

Cited by 4 publications

References 18 publications

Rubella Virus Strain-Associated Differences in the Induction of Oxidative Stress Are Independent of Their Interferon Activation

Rubella Virus Strain-Associated Differences in the Induction of Oxidative Stress Are Independent of Their Interferon Activation

Evaluation of Methods for Duration of Preservation of Rna Quality in Rat Liver Used for Transcriptome Analysis

Validation and application of normalization factors for gene expression studies in rubella virus‐infected cell lines with quantitative real‐time PCR

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