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| Status |
Public on Apr 05, 2008 |
| Title |
Ductal Carcinoma Progression |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
To obtain insight into the molecular basis of ductal carcinoma in situ (DCIS) and its progression by infiltrating surrounding tissue, we have performed cellular-based gene expression analysis of pure DCIS and DCIS with co-existing invasive ductal carcinoma (IDC) and compared the histological and molecular aspects between these morphologically identical lesions seeking to find key genes involved in DCIS progression. For that, 30 samples were evaluated, 4 non-neoplastic (N), 5 pure DCIS, 11 DCIS with co-existing IDC (DCIS-IDC) and 10 IDC. All samples were laser capture microdissected and RNAs were amplified using T7-based methodology. Microarray technology was performed using a customized cDNA platform containing 4,608 human genes. To classify the 4 sample groups according to molecular similarity we performed comparisons of their general expression pattern using ANOVA test (pFDR<0,01) followed by Tukey´s test (fold>l2l). Among the 4 sample groups, as expected, non-neoplastic cells reported the most distinct gene expression pattern. However, among the 3 groups of neoplastic cells, in contrast to morphological aspects, pure DCIS had the most distinct expression profile when compared to the other lesions: DCIS-IDC and IDC. Additionally, by comparison among pure DCIS, DCIS-IDC and N, we identified 147 genes potentially involved in DCIS progression. Unsupervised hierarchical cluster based on expression profile of this gene-set could discriminate DCIS-IDC from 60% of pure DCIS samples.
Keywords: disease state analysis
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| Overall design |
Fresh-frozen human breast samples were retrieved from the Tumor Tissue Biobank of the Medical and Research Center - Hospital A. C. Camargo, São Paulo. This research was approved by Ethic Committee of the Medical and Research Center - Hospital A. C. Camargo under number 587/04 and a written informed consent signed by all participants. All patients had presented at least 5 years of follow-up. Thirty samples were evaluated, 4 non-neoplastic breast samples (MN), 5 pure DCIS (stage 0; DCIS-0), 11 DCIS with co-existing IDC (DCIS) and 10 IDC. The non-neoplastic samples were obtained from perilesional mammary specimens from patients submitted to resection of benign lesions. All cells types were laser capture microdissected using PixCell II LCM system (Arcturus Engineering, Mountain View, CA). The total RNA was extracted by using the PicoPureä RNA Isolation kit (Arcturus Engineering # KT0204). A two-round linear amplification procedure based on T7-driven amplification was carried out. Amplified RNA was then used in a transcriptase reverse reaction into cDNA in the presence of Cy3- or Cy5-labeled dCTP. HB4a normal luminal epithelial mammary cell line (O’Hare et al 1991) was amplified following the same protocol and used as reference for microarray hybridizations. Dye swap was performed for each sample analyzed and used as replicate samples.
For the raw_data (lowess_data), see Web Link below.
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| Contributor(s) |
Castro NP, Osório CT, Torres C, Bastos EP, Soares FA, Brentani HP, Carraro DM |
| Citation(s) |
18928525 |
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| Submission date |
Apr 03, 2008 |
| Last update date |
Mar 05, 2019 |
| Contact name |
Nádia Pereira Castro |
| Organization name |
Hospital AC Camargo
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| Lab |
Genomic and Molecular Biology Laboratory
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| Street address |
R: Prof Antônio Prudente, 211
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| City |
São Paulo |
| State/province |
SP |
| ZIP/Postal code |
01509-900 |
| Country |
Brazil |
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| Platforms (1) |
| GPL1930 |
Homo sapiens 4.8K 02-01 amplified cDNA |
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| Samples (60)
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| Relations |
| BioProject |
PRJNA107021 |
| Supplementary data files not provided |
| Processed data included within Sample table |
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