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• Research Paper Volume 11, Issue 13 pp 4688-4705

  Assessment of ERBB2 and TOP2α gene status and expression profile in feline mammary tumors: findings and guidelines

  Relevance score: 15.812808

  Daniela Ferreira, Maria Soares, Jorge Correia, Filomena Adega, Fernando Ferreira, Raquel Chaves

  Keywords: ERBB2, TOP2α, feline mammary tumors, DNA, RNA, clinicopathological features

  Published in Aging on July 12, 2019

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  In humans, the ERBB2 gene amplification and overexpression are biomarkers for invasive breast cancer and a therapeutic target. Also, TOP2α gene aberrations predict the response to anthracycline-based adjuvant chemotherapy. Although feline mammary tumors (FMTs) are good models in comparative oncology, scarce data is available regarding the ERBB2 and TOP2α status. In this study, and for the first time, the ERBB2 DNA status and RNA levels of intracellular (ICD) and extracellular (ECD) coding regions were compared with TOP2α gene status and expression profile, in samples of FMTs and disease-free tissues from the same animal. Results showed that ERBB2 and TOP2α gene status are highly correlated (r=0.87, p<0.0001, n=25), with few tumor samples presenting amplification. Also, the majority of the FMTs showed ERBB2 overexpression coupled with TOP2α overexpression (r=0.87, p<0.0001, n=27), being the ERBB2-ICD and ECD transcripts highly correlated (r=0.97, p<0.0001, n=27). Significant associations were found between TOP2α gene status or ERBB2 and TOP2α RNA levels with several clinicopathological parameters. This work highlights the need of experimental designs for a precise evaluation of ERBB2 and TOP2α gene status and its expression in FMTs, to improve their clinical management and to further validate them as a suitable model for comparative oncology studies.

  

  ERBB2 and TOP2α maintain the copy number in feline mammary tumors. (a-b) Fold change of ERBB2 (a) and TOP2α (b) DNA copy number in feline mammary tumors (FMT) analyzed by real-time qPCR and compared with a disease-free tissue (DFT) sample collected from the same animal (control). The percentage of tumors showing an increased, maintained or decreased gene copy number of ERBB2 (a) and TOP2α (b) is presented in the upper right corner of each graph. Values are mean ± SD of three replicates. *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001 were determined by Student’s t-test.

  
  
  

  

  ERBB2 and TOP2α DNA and RNAs correlogram. Correlation among ERBB2 and TOP2α DNA and RNAs. This correlogram was obtained using the R software. As some analysis presented a different “n”, the data was simultaneously analyzed in GraphPad software and the r-values were corrected by the GraphPad values.

  
  
  

  

  The RNA levels of the EBB2 ICD and ECD and TOP2α are altered in the majority of the FMTs. Fold change of Erbb2 ICD (a) and ECD (b) RNA regions and TOP2α RNA (c) quantified by real-time RT-qPCR in FMTs and compared with disease-free tissue collected from the same donor. The percentage of tumors with an increase, maintenance or decrease in the ERBB2 ICD (a), ECD (b) and TOP2α (c) RNA levels is also presented, in the upper right corner of each graph. Values are mean ± SD of three replicates. *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001 are determined by Student’s t-test.

  
  
  

  

  Statistical associations between TOP2α gene status and lymphovascular invasion (a) and between ERBB2 RNAs levels and oral contraceptive administration (b). The data are presented as box-plot graphics, showing median, quartiles and extreme values for each category. The p-value was obtained by one-way analysis of variance test (ANOVA, Tukey Post Hoc Multiple Comparisons).