Heme status affects human hepatic messenger RNA and microRNA expression

Table 2 Summary of data analysis with use of ingenuity pathway

	6 h		24 h
Name	P value	Ratio	P value	Ratio
Top canonical pathways
Nrf2-mediated oxidative stress response	2.64 × 10-11	28/193 (0.145)	1.58 × 10-9	61/193 (0.316)
Protein ubiquitination pathway	1.44 × 10-9	31/274 (0.133)	3.26 × 10-8	75/274 (0.274)
Aldosterone signaling in epithelial cells	8.31 × 10-8	21/170 (0.124)	-	-
Glucocorticoid receptor signaling	1.6 × 10-5	24/295 (0.081)	-	-
P53 signaling	1.75 × 10-3	10/96 (0.104)	-	-
Biosynthesis of steroids	-	-	3.22 × 10-7	14/121 (0.116)
Propanoate metabolism	-	-	1.45 × 10-6	24/121 (0.198)
Urea cycle and metabolism of amino groups	-	-	5.8 × 10-6	16/78 (0.205)
Top toxicity lists
Nrf2-mediated oxidative stress response	3.03 × 10-11	30/237 (0.127)	5.41 × 10-9	66/237 (0.278)
Renal necrosis/cell death	1.05 × 10-4	25/314 (0.08)	1.06 × 10-5	79/314 (0.252)
Liver necrosis/cell death	1.79 × 10-3	14/166 (0.084)	-	-
P53 signaling	2.06 × 10-3	10/95 (0.105)	1.54E × 10-4	30/95 (0.316)
Liver proliferation	2.54 × 10-3	12/133 (0.09)	-	-
Cholesterol biosynthesis	-	-	2.26 × 10-13	16/16 (1)
FXR/RXR activation	-	-	5.84 × 10-4	26/86 (0.302)

P values are for comparison of results following treatment with iron protoporphyrin (heme) vs 4, 6-dioxoheptanoic acid (DHA). Listing of all microRNAs with ≥ 1.5 fold differences in expression at 6 h after treatment of Huh-7 cells with heme (10 μmol/L) vs DHA (500 μmol/L). Cells were cultured, treated, harvested and total RNA prepared and assays as described in Materials and Methods. FXR/RXR: Farnesoid X receptor/retinoic X receptor; Nrf2: Nuclear erythroid-2 related factor.