The regulation of β-catenin’s expression in HEK293T and Hela cells by LH14E, cisplatin and mitomycin C
Authors:Christopher Ott, Nilay Patel, Thu Tran, Chiaokai Wen
Mentor:Nilay Patel, Ph. D. Assistant Professor, Department of Biological Science, California State University, Fullerton
β-catenin protein plays as a transcriptional coactivator of transcriptional factor of genes that regulate cell proliferation. Other researches show that mutations which cause level of β-catenin protein increasing relate to many cancers such as breast cancer, colorectal cancer, melanoma, prostate cancer, lung cancer. Therefore those drugs that can reducing β-catenin level have anti-cancer potential. LH14E was newly synthesized by Dr. Peter de Lijser’s lab, CSUF. Some initial experiments have shown that LH14E reduces β-catenin protein level. This project is to use various methods to examine the effect and learn some about affecting mechanism of LH14E, as well as mitomycin C and cisplatin, on β-catenin expression. Mitomycin C and cisplatin have been used as anti-cancer drugs, but it is not known whether they affect β-catenin expression. We hypothesized that mitomycin C, cisplatin and LH 14E reduce beta-catenin mRNA stability. qPCR was used to test the relative expression of β-catenin regarding to the three drug treatments. Immunocytochemistry (ICC), western blotting, and TOPflash assay were also employed to study the protein levels of β-catenin by the three drugs. qPCR results were consistent that the three drugs do not change β-catenin mRNA expression. All ICC, western blotting and TOPflash assay show that LH14E clearly reduces level of active β-catenin protein. However, the results vary with cisplatin and mitomycin C. In conclusion, LH14E reduces β-catenin expression but not at mRNA level. The effect of cisplatin and mitomycin C on β-catenin level needs more research for solid conclusion.