Assessment of Gene Expression Profiles in Peripheral Occlusive Arterial Disease
2012
Abstract Background Molecular events responsible for the onset and progression of peripheral occlusive arterial disease (POAD) are incompletely understood. Gene expression profiling may point out relevant features of the disease. Methods Tissue samples were collected as operatory waste from a total of 36 patients with (n = 18) and without (n = 18) POAD. The tissues were histologically evaluated, and the patients with POAD were classified according to Leriche-Fontaine (LF) classification: 11% with stage IIB, 22% with stage III, and 67% with stage IV. Total RNA was isolated from all samples and hybridized onto Agilent 4×44K Oligo microarray slides. The bioinformatic analysis identified genes differentially expressed between control and pathologic tissues. Ten genes with a fold change ≥ 2 (1 with a fold change ≥ 1.8) were selected for quantitative polymerase chain reaction validation ( GPC3 , CFD , GDF10 , ITLN1 , TSPAN8 , MMP28 , NNMT , SERPINA5 , LUM , and FDXR ). C-reactive protein (CRP) was assessed with a specific assay, while nicotinamide N-methyltransferase (NNMT) was evaluated in the patient serum by enzyme-linked immunosorbent assay. Results A multiple regression analysis showed that the level of CRP in the serum is correlated with the POAD LF stages ( r 2 = 0.22, P = 0.046) and that serum NNMT is higher in IV LF POAD patients ( P = 0.005). The mRNA gene expression of LUM is correlated with the LF stage ( r 2 = 0.45, P = 0.009), and the mRNA level of ITLN1 is correlated with the ankle-brachial index ( r 2 = 0.42, P = 0.008). Conclusions Our analysis shows that NNMT , ITLN1 , LUM , CFD , and TSPAN8 in combination with other known markers, such as CRP, could be evaluated as a panel of biomarkers of POAD.
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