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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.
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BMC Cancer

ISSN: 1471-2407

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