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| Author(s): |
Helvia Nascimento
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas |
| Defense date: | 2007-02-28 |
| Examining board members: |
Carmen Silvia Passos Lima;
Benedito Mauro Rossi;
Andre Luiz Vettore;
João Jose Fagundes;
José Vassallo
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| Advisor: | Carmen Silvia Passos Lima; Fernando Ferreira Costa |
| Abstract | |
The molecular mechanisms involved in sporadic colon adenocarcinoma (SCA) are still not completely elucidated. Recently, the serial analysis of gene expression (SAGE) method has allowed the global analysis of genes expressed in diverse cellular types but there are no studies in purified epithelial cells of SCA moderately differenciated. We have characterized through SAGE the global gene expression of neoplastic epithelial cells from a SCA moderately differenciated patient (SAGE CC) and normal epithelial cells from a megacolon patient (SAGE CN). After automatic sequencing, a total of 44.004 tags from SAGE CC and 43.570 tags from SAGE CN profiles were generated, representing 16.484 and 13.479 unique tags, respectively. Comparing both profiles, 171 differentially expressed transcripts were identified (P< 0.001; fold = 5), including 10.0% that may represent novel transcripts. The expression of 10 selected genes was further investigated by realtime polymerase chain reaction (qPCR) in the SCA moderately differenciated epithelial cells sample (SAGE CC), with the purpose of to validate the results obtained by the SAGE method, and also in five epithelial cells samples from the same type of SCA patients. Similar expressions were seen in 80% (CEACAM6, KLK6, LYZ, PFN1, S100A8, S100A9, VIL2 e ZFHX1B) and discordant expressions were seen in 20% (PLA1A e ZNF277) of analysed genes. On SAGE CC sample and samples of the SCA patients, all genes presented similar expressions measured by both methods. We observed abnormal expression of genes involved with cell proliferation and differentiation, and with response to stress in neoplastic epithelial cells. Also, were found abnormal expressions of genes not related with the disease and not identified genes. Together, our results may contribute for the identification of genes involved in the origin or progression of SCA moderately differenciated, as well as for the discovery of new therapeutical agents, with specific action on abnormal proliferation of the neoplastic cells. (AU) | |