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This dissertation, Role of AMP-activated Protein Kinase in Cervical Cancer Cell Growth by Yee-man, Yu, 余綺雯, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled Role of AMP-activated Protein Kinase in Cervical Cancer Cell Growth submitted by Yu Yee Man for the degree of Master of Philosophy at The University of Hong Kong in August 2006 Objectives: Tumors are usually grown in a nutrient-deprived environment due to insufficient vascularization. AMP-activated protein kinase (AMPK) has been well recognized as an important mediator of stress signals, including hypoxia and nutrient deprivation. We have previously identified gene amplification of the catalytic alpha 1 subunit of AMPK by comparative genome hybridization in cervical tumors. Also, both mRNA and protein of AMPKα1 were found to be over-expressed in cervical tumors. This study aims to investigate the effect of AMPK activation on the growth of cervical cancer cells. We also look at the expression of LKB1 (a known upstream activator of AMPK) in cervical cancer cells and the involvement of calcium-dependent protein kinase kinase, CaMKK, (another potential upstream activator of AMPK) in activating AMPK. Methods: The alterations of expression of AMPK subunits of five cervical cancer cell lines (HeLa, SiHa, CaSki, C41 and C33A) under glucose deprivation or hypoxia were studied by cDNA microarray anaylsis. Besides, HeLa and SiHa, were treated with either glucose supply or glucose deprived conditions; and with or without AMPK activator, AICAR. The cell survival was measured by XTT assay and confirmed by flow cytometry. We also did PCR to study LKB1 gene expression in cervical cancer cells. In addition, the Isame set of experiments described above was repeated with another drug A23187 which, by activating CaMKK, may also be a potential AMPK activator. Results: We found that AMPKα1 and AMPKγ1 mRNA levels were increased after glucose deprivation or hypoxia treatment. Besides, under glucose deprivation, the cell survival of HeLa treated with AICAR was more than control at 48 hours post drug treatment. Similar results were observed in SiHa. On the other hand, with glucose supply, HeLa cell growth was inhibited under AICAR treatment compared to that without AICAR treatment. The data on cell proliferation assay was confirmed by flow cytometry. In addition, we found that LKB1 was not expressed in HeLa and SiHa due to gene deletion. Therefore we repeated the above experiments with A23187, a CaMKK activator. Like AICAR, A23187 was also able to activate AMPK, resulting in reduced cell death in HeLa and SiHa under glucose deprivation. Moreover, it also induced the cell death under glucose supply. The data indicates that both AICAR and A23187 had similar effects on cervical cancer cell growth via AMPK signaling. Conclusions: These data suggest that AMPKα1 and AMPKγ1 were both up-regulated at mRNA level in cervical cancer cells under hypoxia or glucose deprivation. Moreover, the activation of AMPK by both AICAR and A23187 inhibited cell growth with glucose supply but reduced cell death under glucose deprivation. We therefore suggest that AMPK may have a dual role in controlling cervical cancer cell growth. Moreover, AMPK activation in HeLa and SiHa is LKB1 independent but instead depends on CaMKK, an alternative upstream activator of AMPK. II DOI: 10.5353/th_b3745997 Subjects: Protein kinasesCancer cells - GrowthCervix uteri - Cancer - Molecular aspects

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