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BMC Cancer 2008
Gene expression variation between distinct areas of breast cancer measured from paraffin-embedded tissue coresAbstract: In this study we used tissue cylinders as known from the construction of tissue microarrays. RNA was isolated with a robust protocol recently developed for RNA derived from FFPE material. Gene expression was measured by quantitative reverse transcription PCR.Sixteen tissue blocks from 7 patients diagnosed with multiple histological subtypes of breast cancer were available for this study. After verification of appropriate localization, sufficient RNA yield and quality, 30 tissue cores were available for gene expression measurement on TaqMan? Low Density Arrays (16 invasive ductal carcinoma (IDC), 8 ductal carcinoma in situ (DCIS) and 6 normal tissue), and 14 tissue cores were lost. Gene expression values were used to calculate scores representing the proliferation status (PRO), the estrogen receptor status and the HER2 status. The PRO scores measured from entire sections were similar to PRO scores determined from IDC tissue cores. Scores determined from normal tissue cores consistently revealed lower PRO scores than cores derived from IDC or DCIS of the same block or from different blocks of the same patient.We have developed optimized protocols for RNA isolation from histologically distinct areas. RNA prepared from FFPE tissue cores is suitable for gene expression measurement by quantitative PCR. Distinct molecular scores could be determined from different cores of the same tumor specimen.Diagnosis and prognosis of breast cancer are still mainly based on clinical, histological and immunohistochemical parameters, which are at best semi-quantitative [1,2]. Recently, molecular characterization of breast cancer has greatly increased the understanding of biological pathways that are altered during neoplastic transformation. Molecular markers have a great impact on elucidating the biological variance within tumors, they allow new and more accurate classifications and they have the potential to improve diagnosis, estimation of prognosis and treatment decisions in individua
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