Heme status affects human hepatic messenger RNA and microRNA expression

Table 1 Confirmation and comparison of selected messenger RNA array results with those of quantitative real time polymerase chain reaction for selected genes

	Array		qRT-PCR
Gene	Fold change	P value	Fold change	P value
Heme vs DHA-6 h time point
HMOX1	59.9	9.97 × 10-9	87.9	10-35
IL-8	20.50	3.92 × 10-5	13.7	0.016
ANXA1	7.18	1.87 × 10-5	32.3	0.015
BACH1	3.04	3.26 × 10-5	3.1	0.0019
Heme vs DHA-24 h time point
HSPB8	77.1	2.66 × 10-6	92	0.010
ANXA1	45.7	5.98 × 10-6	138	0.015
IL-8	33.2	4.19 × 10-5	31.7	0.016
HMOX1	25.6	2.50 × 10-5	27.6	1.33 × 10-7
BACH2	4.50	7.24 × 10-5	22.6	0.011
PNPLA3	-6.48	1.74 × 10-4	-6.65	4.59 × 10-7

P values are for comparison of results following treatment with iron protoporphyrin (heme) vs 4, 6-dioxoheptanoic acid (DHA). Listing of all messenger RNAs with ≥ 1.5 fold differences in expression at 6 h, 24 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. qRT-PCR: Quantitative real time polymerase chain reaction; HMOX1: Heme oxygenase 1; IL-8: Interleukin-8; ANXA1: Annexin A1; BACH: Basic leucine zipper transcription factor; HSPB8: Heat shock 22kDa protein 8; PNPLA3: Patatin-like phospholipase domain-containing protein 3.