Gene expression variation between distinct areas of breast cancer measured from paraffin-embedded tissue cores

BACKGROUND: Diagnosis and prognosis in breast cancer are mainly based on histology and immunohistochemistry of formalin-fixed, paraffin-embedded (FFPE) material. Recently, gene expression analysis was shown to elucidate the biological variance between tumors and molecular markers were identified that led to new classification systems that provided better prognostic and predictive parameters. Archived FFPE samples represent an ideal source of tissue for translational research, as millions of tissue blocks exist from routine diagnostics and from clinical studies. These should be exploited to provide clinicians with more accurate prognostic and predictive information. Unfortunately, RNA derived from FFPE material is partially degraded and chemically modified and reliable gene expression measurement has only become successful after implementing novel and optimized procedures for RNA isolation, demodification and detection. METHODS: 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. RESULTS: 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(R) 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. CONCLUSION: 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.

Educational Systems and Gender Segregation in Education: A Three-Country Comparison of Germany, Norway and Canada

How do institutional settings and their embedded policy principles affect gender-typed enrolment in educational programmes? Based on gender-sensitive theories on career choice, we hypothesised that gender segregation in education is higher with a wider range of offers of vocational programmes. By analysing youth survey and panel data, we tested this assumption for Germany, Norway and Canada, three countries whose educational systems represent a different mix of academic, vocational and universalistic education principles. We found that vocational programmes are considerably more gender-segregated than are academic (e.g. university) programmes. Men, more so than women, can avoid gender-typed programmes by passing on to a university education. This in turn means that as long as their secondary school achievement does not allow for a higher education career, they have a higher likelihood of being allocated to male-typed programmes in the vocational education and training (VET) system. In addition, social background and the age at which students have to choose educational offers impact on the transition to gendered educational programmes. Overall, gender segregation in education is highest in Germany and the lowest in Canada. We interpret the differences between these countries with respect to the constellations of educational principles and policies in the respective countries.