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Table 2 Relationships between mouse (m) and human (h) mRNA levels in the 150 human tumor xenografts

From: Vasculature analysis of patient derived tumor xenografts using species-specific PCR assays: evidence of tumor endothelial cells and atypical VEGFA-VEGFR1/2 signalings

  hCD31 mCd31 hCD105 mCd105 hVEGFR1 mVegfr1 hVEGFR2 mVegfr2 hVEGFA mVegfa
mCd31 0.0251          
  0.762          
hCD105 0.043 0.121         
  0.60 0.14         
mCd105 0.040 0.928 0.189        
  0.63 <0.0000001 0.02        
hVEGFR1 0.065 0.022 -0.076 0.004       
  0.43 0.79 0.35 0.96       
mVegfr1 0.076 0.851 0.305 0.877 0.006      
  0.35 <0.0000001 <0.0002 <0.0000001 0.94      
hVEGFR2 0.010 -0.029 0.232 -0.036 -0.036 0.070     
  0.91 0.72 <0.005 0.66 0.66 0.40     
mVegfr2 0.003 0.912 0.173 0.919 -0.017 0.858 -0.090    
  0.98 <0.0000001 <0.05 <0.0000001 0.83 <0.0000001 0.27    
hVEGFA 0.095 0.477 0.319 0.563 0.090 0.726 0.131 0.517   
  0.25 <0.0000001 <0.0002 <0.0000001 0.27 <0.0000001 0.11 <0.0000001   
mVegfa 0.031 0.505 0.194 0.524 0.304 0.514 0.062 0.413 0.328  
  0.70 <0.0000001 <0.05 <0.0000001 <0.0002 <0.0000001 0.45 <0.0000001 <0.00005  
% mouse cells -0.016 0.828 0.113 0.865 0.154 0.715 -0.145 0.797 0.364 0.666
  0.84 <0.0000001 0.17 <0.0000001 0.06 <0.0000001 0.08 <0.0000001 <0.000005 <0.0000001
  1. Results, expressed as N-fold differences in target gene expression relative to the mouse and human TBP genes (both the mouse and human TBP transcripts) and termed “Ntarget”, were determined as Ntarget = 2∆Ctsample , where the ∆Ct value of the sample was determined by subtracting the average Ct value of target gene (human or mouse) from the average Ct value of ‘Total-TBP’ gene). The Ntarget values of the tumor samples were subsequently normalized such that the value for mRNA level was 1 when Ct=35. Target mRNA levels that were total absence or very low (Ct > 38) in tumor samples were scored ‘0’ for non expressed. As for calculation of % of mouse cells, specific mouse Tbp gene expression and the expression of both the mouse and the human TBP genes were studied by real-time qRT-PCR using the mouse Tbp as target gene and the ‘Total-TBP’ as endogenous RNA control. Results, expressed as N-fold differences in specific mouse Tbp gene expression (using mouse Tbp primers) relative to the sum of the mouse and the human TBP gene expression (using ‘Total-TBP’ primers), termed NMm-TBP, are determined by theformula: NMm-TBP = 2DCtsample. The DCt value of the sample is determined by subtracting the Ct value of the mouse TBP gene from the Ct value of the Total TBP gene. The NMm-TBP values of the samples are subsequently normalized such that the median of NMm-TBP values of 4 mouse tissues was 100. As TBP is a ubiquitously expressed housekeeping gene, showing similar expression in our human and mouse tissues (Ct=27 for 5 ng cDNA), the final result (normalized NMm-TBP value) gives an estimate of the proportion of mouse cell content for a given xenograft. 1Spearman correlation coefficient, 2p value of Spearman rank correlation test, in bold when p is significant.