Housekeeping gene stability in pristane-induced arthritis and antigen-induced pulmonary inflammation of rats

Jiang, Congshan; Meng, Liesu; Zhu, Wenhua; Shahzad, Muhammad; Yang, Xudong; Lu, Shemin

doi:10.1007/s00011-009-0027-5

Cited by 8 publications

(4 citation statements)

References 21 publications

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“…We would therefore recommend that for gene expression studies investigating the pathogenesis of mesothelium adhesions, ACTB, YWHAZ, or CYC1 be used as the internal control. These findings are consistent with recent studies indicating that both the experimental conditions and the presence of pathology can have an impact on the expression of these commonly used internal controls (18)(19)(20). The assumption that HKG are equally stable under all conditions must therefore be further questioned.…”

Section: Discussionsupporting

confidence: 92%

Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium

Sadek

Cagampang

Bruce

et al. 2012

Fertility and Sterility

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Section: Discussionsupporting

confidence: 92%

Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium

Sadek

Cagampang

Bruce

et al. 2012

Fertility and Sterility

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“…Using PPIA in pair with other HKG has frequently been recommended in CRC studies. 20 Accordingly, it was top-ranked also in our study, less so by Norm-Finder and in a small bowel. 20 Accordingly, it was top-ranked also in our study, less so by Norm-Finder and in a small bowel.…”

Section: Discussionsupporting

confidence: 53%

Expression Stability of Common Housekeeping Genes Is Differently Affected by Bowel Inflammation and Cancer

Krzystek−Korpacka¹,

Diakowska

Bania

et al. 2014

Inflammatory Bowel Diseases

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Instability of housekeeping genes (HKG), supposedly unregulated and hence used as normalizers, may dramatically change conclusions of quantitative PCR experiments. The effect of bowel inflammation on HKG remains unknown. Expression stability of 15 HKG (ACTB, B2M, GAPDH, GUSB, HPRT1, IPO8, MRPL19, PGK1, PPIA, RPLP0, RPS23, SDHA, TBP, UBC, and YWHAZ) in 166 bowel specimens (91 normal, 35 cancerous, and 40 inflamed) was ranked by coefficients of variation (CV%) or using dedicated software: geNorm and NormFinder. The RPS23, PPIA, and RPLP0 were top-ranked, whereas IPO8, UBC and TBP were the lowest-ranked HKG across inflamed/cancerous/normal colonic tissues. The pairs RPS23/RPLP0, PGK1/MRPL19, or PPIA/RPLP0 were optimal reference by CV%, NormFinder, and geNorm, respectively. Colon inflammation affected HKG more pronouncedly than cancer with ACTB significantly down- and B2M upregulated. In inflammatory bowel disease (IBD), different genes were top-ranked in a large and small bowel, whereas TBP, UBC, and IPO8 were lowest-ranked in both. For patients with IBD at large, RPS23/PPIA, PGK1/MRPL19, and PPIA/RPLP0 were found optimal by CV%, NormFinder, and geNorm, respectively. ACTB and B2M expression was related to CRC stage and positively correlated with clinical activity of IBD. Although GAPDH was upregulated neither in CRC nor IBD, it tended to positively correlate with tumor depth and Crohn's disease activity index. Normalizing against GAPDH affected experimental conclusions in a small but not large bowel. Bowel inflammation significantly affects several classic HKG. The pair PPIA/RPLP0 is a common optimal reference for studies encompassing tissues sampled from colorectal cancer and IBD patients. Using ACTB or B2M is not recommended.

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“…The TIMP-3 mRNA as measured by quantitative RT-PCR remained at a constant level within the tested time period in the sham group as shown in Figure 1(the data for TIMP-3 RT-PCR represents values normalized to GAPDH, while GAPDH was maintained as a relative constant throughout the study [Thellin et al, 1999;Jiang et al, 2009;Aswal et al, 2008]). At 4 h post-TBI, the amount of TIMP-3 mRNA (in both the ipsilateral and contralateral cortex and ipsilateral and contralateral hippocampus) in the normothermic group was significantly increased compared to the sham group ( p < 0.01).…”

Section: Rt-pcrmentioning

confidence: 86%

The Effect of Hypothermia on the Expression of TIMP-3 after Traumatic Brain Injury in Rats

Feng

Mao

Liang

et al. 2014

Journal of Neurotrauma

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Here we investigate the effect of hypothermia on the expression of apoptosis-regulating protein TIMP-3 after fluid percussion traumatic brain injury (TBI) in rats. We began with 210 adult male Sprague-Dawley rats and randomly assigned them to three groups: TBI with hypothermia treatment (328C), TBI with normothermia (378C), and sham-injured controls. TBI was induced by a fluid percussion TBI device. Mild hypothermia (328C) was achieved by partial immersion in a water bath (08C) under general anesthesia for 4 h. The rats were killed at 4, 6, 12, 24, 48, and 72 h and 1 week after TBI. The mRNA and protein level of TIMP-3 in both the injured and uninjured hemispheres of the brains from each group were measured using RT-PCR and Western blotting. In the normothermic group, TIMP-3 levels in both the injured and uninjured hemispheres were significantly increased after TBI compared with those of sham-injured animals ( p < 0.01). In contrast, post-traumatic hypothermia significantly attenuated this increase. According to the RT-PCR and Western blot analyses, the maximum mRNA levels of TIMP-3 were reduced to 60.60 AE 2.30%, 55.83 AE 1.80%, 66.03 AE 2.10%, and 64.51 AE 1.50%, respectively, of the corresponding values in the normothermic group in the injured and uninjured hemispheres (cortex and hippocampus) of the hypothermia group ( p < 0.01), while the respective maximum protein levels of TIMP-3 were reduced to 57.50 AE 1.50, 52.67 AE 2.20, 60.31 AE 2.50 and 54.76 AE 1.40 ( p < 0.01). Our data suggest that moderate fluid percussion brain injury significantly upregulates TIMP-3 expression, and that this increase may be suppressed by hypothermia treatment.

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Housekeeping gene stability in pristane-induced arthritis and antigen-induced pulmonary inflammation of rats

Abstract: We suggest that the stable gene mentioned above or geometric mean of three best references in each experimental condition should be used as the reference and the variable ones should be avoided in similar study designs.

Cited by 8 publications

References 21 publications

Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium

Variation in stability of housekeeping genes in healthy and adhesion-related mesothelium

Expression Stability of Common Housekeeping Genes Is Differently Affected by Bowel Inflammation and Cancer

The Effect of Hypothermia on the Expression of TIMP-3 after Traumatic Brain Injury in Rats

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