Inhibition of RE‐1 silencing transcription factor (REST) inhibits survival and proliferation of primary hepatocytes under the influence of hepatocye growth factor (HGF) and epidermal growth factor (EGF)

Abstract

The objective of the present work was to test the expression and role of REST in primary hepatocytes under the influence of growth factors HGF and EGF. REST (also known as neuron restriction silencing factor NRSF) has previously been shown to maintain self renewal and pluripotency in mouse embryonic stem cells by maintaining the expression of self renewal regulators like Oct3/4, Nanog, cMyc, and Klf4. Primary rat hepatocytes undergo dedifferentiation and proliferation in the presence of hepatocyte growth medium (containing HGF and EGF). Primary hepatocytes were cultured with or without HGF and EGF. Plates were harvested at 0, 2, 4, 6, 8, and 10 days for mRNA and protein. REST mRNA and protein were upregulated along with stem cell markers like Oct3/4, Nanog, cMyc, and Klf4 in the presence of HGF and EGF. When REST was inhibited in these hepatocytes using shRNA for REST, these hepatocytes did not proliferate and died as compared to cells treated with the shRNA of luciferase, suggesting that REST expression is important for survival and proliferation of primary hepatocytes under the influence of HGF and EGF. The expression of stem cell markers in the REST inhibited hepatocytes is yet to be determined. This work is supported by 5R01CA103958‐05 titled HGF and signaling pathways in hepatic tissue assembly.