Masters Thesis

Lh14e's Effects are Mediated through P21 and Related Proteins

A small library of compounds synthesized at California State University, Fullerton, were screened for their anti-proliferative activity in three human cell lines: HeLa, HUTU80, and MG63. CyQUANT cell proliferation assay also showed some cell line specificity of best performing compounds. This project focuses on identification of the effects of the compound, LH14E, on p21, a cell cycle regulatory protein. Expression of CDKN1A (p21 gene) is regulated by p53 and KLF6, both of which attenuate cell proliferation. Our results show that LH14E increases expression of CDKN1A and the amount of p21 protein. Luciferase reporter gene assay revealed an increase in the transcriptional activity of the p21 promoter upon treatment with LH14E. Western Blot reveals p53 protein levels are not affected by treatment with LH14E, but KLF6 protein increases upon treatment with LH14E. These results suggest that LH14E activates KLF6 expression, and subsequently increases p21 expression, which together may cause cell cycle arrest. These analyses will help identify mechanism of action for the hit compounds in this library of compounds.

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