Apoptosis – associated genes and their role in predicting responses to neoadjuvant breast cancer treatment
Daniel Tvrdík, Helena Skálová, Pavel Dundr, Ctibor Povýšil, Zuzana Velenská, Adéla Berková, Libor Staněk, Luboš Petruželka
Med Sci Monit 2012; 18(1): BR60-67
Available online: 2011-12-22
Background: Neoadjuvant chemotherapy is used in the treatment of breast carcinoma because it substantially reduces the size of the primary tumor and lymph node metastases. The present study investigated biomarkers that can predict a pathologic response to the therapy.
Material/Methods: The role of apoptosis in regression of the tumors after neoadjuvant chemotherapy was determined by TUNEL and anti-active caspase 3 assay. The transcriptional profile of 84 key apoptosis genes was evaluated in both pre-therapeutically obtained tumor tissue by core needle biopsy and in specimens removed by final surgery, using a pathway-specific real-time PCR assay. Obtained data were analyzed by hierarchical cluster analysis and correlation analysis. The immunohistochemical profile of each tumor was determined using the standard ABC method.
Results: On the basis of a hierarchical cluster analysis of 13 significantly changed genes, we divided patients into good and poor prognosis groups, which correlate well with progression-free survival. In the good prognosis group, we found a statistically significant down-regulation of the expression of MCL1 and IGF1R genes after neoadjuvant treatment. We also found a statistically significant overexpression of BCL2L10, BCL2AF1, CASP8, CASP10, CASP14, CIDEB, FADD, HRK, TNFRSF25, TNFSF8 and CD70 genes. In contrast, we found up-regulation of IGF1R after the treatment in the group with poor prognosis.
Conclusions: Gene expression profiling using real-time PCR assay is a valuable research tool for the investigation of molecular markers, which reflect tumor biology and treatment response.
Keywords: In Situ Nick-End Labeling, Humans, Gene Expression Profiling - methods, Female, Cluster Analysis, Chemotherapy, Adjuvant, Breast Neoplasms - drug therapy, Apoptosis Regulatory Proteins - metabolism, Apoptosis - drug effects, Antineoplastic Agents - pharmacology, Middle Aged, Neoadjuvant Therapy, Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction - methods