Hepatocellular carcinoma (HCC) is a common form of cancer with prevalence worldwide. There are many factors that lead to the development and progression of HCC. This study aimed to identify potential new tumor suppressors, examine their function as cell cycle modulators, and investigate their impact on the cyclin family of proteins and cyclin-dependent kinases (CDKs). In this study, the pyruvate dehydrogenase kinase (PDK)4 gene was shown to have potential tumor suppressor characteristics. PDK4 expression was significantly downregulated in human HCC. Pdk4−/− mouse liver exhibited a consistent increase in cell cycle regulator proteins, including cyclin D1, cyclin E1, cyclin A2, some associated CDKs, and transcription factor E2F1. PDK4-knockdown HCC cells also progressed faster through the cell cycle, which concurrently expressed high levels of cyclins and E2F1 as seen in the Pdk4−/− mice. Interestingly, the induced cyclin E1 and cyclin A2 caused by Pdk4 deficiency was repressed by arsenic treatment in mouse liver and in HCC cells. E2f1 deficiency in E2f1−/− mouse liver or knockdown E2F1 using small hairpin RNAs in HCC cells significantly decreased cyclin E1, cyclin A2, and E2F1 proteins. In contrast, inhibition of PDK4 activity in HCC cells increased cyclin E1, cyclin A2, and E2F1 proteins. These findings demonstrate that PDK4 is a critical regulator of hepatocyte proliferation via E2F1-mediated regulation of cyclins.
- Received August 31, 2016.
- Accepted December 9, 2016.
This research was supported by the National Institutes of Health National Institute of Environmental Health Sciences [Grant R01-ES025909], the National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [R01-DK080440, R01-DK104656, and P30-DK34989 (to Yale Liver Center)], the National Institutes of Health National Institute on Alcohol Abuse and Alcoholism [Grants R21-AA022482 and R21-AA024935], and the U.S. Department of Veterans Affairs [Merit Award 1I01BX002634].
- Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics