Category BI L10 Identifying Novel Mechanisms of Cytochrome P450 2E1 regulation

Abstract

Cyp2E1 is a vital enzyme involved in the metabolism of many carcinogens

within the liver. Cyp2E1 over expression, seen in many pathophysiological

condition (diabetes, viral hepatitis, obesity, etc.), is known to increase risk

for hepatocellular carcinoma. This study aimed to elucidate mechanisms by

which Cyp2E1 disregulation occurs in diabetics. Specifically, the effects

of insulin and diabetic medication metformin on Cyp2E1 expression were

studied. Primary rat hepatocytes were treated with appropriate

substances at variable concentrations. Gene and miRNA expression was

analyzed through qRT-PCR while protein expression was analyzed with

immunoblots. Inhibitors/activators were used to identify functional elements

of the regulatory pathway. The expression of miRNA’s linked to the

development of cancer was quantified.

Insulin was shown to decrease Cyp2E1 expression while metformin

induced its expression. Insulin mediated regulation was dependant on PI3-

Kinase, Akt, AMPK, and mTOR, with evidence that the FOXO proteins

were involved. miRNA 212 and 132 were both regulated by insulin (up)

and metformin (down). In silico analysis showed significant homology

between the regulatory and coding regions of the human and rat gene,

supporting extrapolation of data to humans.

This study provides the first detailed information of the specific

mechanisms behind insulin mediated Cyp2E1 regulation and was the first

to identify metformin’s effects on Cyp2E1 expression, results with great

potential for furthering our understanding of diabetes and its relation to

cancer. In addition, insulin and metformin were shown to regulate the

expression of miRNA’s related to hepatocellular carcinoma, providing novel

evidence that they are involved in epigenetic regulation.

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H3K4me3, Examining. "Identification of Preferential Target Sites for Human

DNA Methyltransferases." Oxford Journals | Life Sciences | Nucleic Acids
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