ABSTRACT
Hepatocellular carcinoma (HCC) is one of the most prevalent cancers in Taiwan. The development of hepatocellular carcinoma is a multi-step process associated with alterations in genes expression such as activation of oncogenes and inactivation of tumor suppressor genes. Mutation/deletion of tumor suppressor gene p53 occurs in 40-50% HCC. Moreover, patients with p53 inactivation have significantly shorter survival after surgery. Inactivation of p53 leads to chromosome instability and may alter expression of its downstream target genes including 14-3-3s for cell cycle arrest or PUMA for apoptosis induction. In this thesis study, we employed five human hepatoma cell lines and ten surgical HCC samples containing paired normal and tumor tissues to investigate 14-3-3s and PUMA expression during liver carcinogenesis.
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) revealed that 14-3-3s mRNA expression was detected in well and poorly differentiated hepatoma cells except Mahlavu cells. Western blot analysis further validated such finding that 14-3-3s protein is not detectable in Mahlavu cell. In human surgical HCC tissues, qRT-PCR showed that 14-3-3s mRNA was elevated in 90% of HCC tissues. Western blot analysis indicated that 14-3-3s protein level was increased in 60% of HCC tissues. Finally, immunohistochemical analysis revealed that 14-3-3s was up-regulated in 50% of HCC tissues comparing with their adjacent non-tumor tissues. Together, these results indicated that 14-3-3s expression was up-regulated in HCC.
qRT-PCR and western blot analysis indicated that PUMA mRNA and protein levels were decreased in human and rat hepatoma cells. In human surgical HCC tissues, qRT-PCR showed that PUMA mRNA was reduced in 60% of HCC tissues. Western blot analysis indicated that PUMA protein level was decreased in 100 of HCC tissues. Finally, immunohistochemical analysis revealed that PUMA was down-regulated in 70% of HCC tissues comparing with their adjacent non-tumor tissues. Together, these results indicated that PUMA expression was down-regulated in HCC.
In the future, large-scale analysis using more HCC samples will be required to delineate the correlation of 14-3-3s/PUMA expression with clinical parameters of HCC.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0214105-143433 |
| Date | 14 February 2005 |
| Creators | Ho, Cheng-lei |
| Contributors | none, none, nono, non |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0214105-143433 |
| Rights | not_available, Copyright information available at source archive |
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