To substantiate the microarray results, qRT-PCR was performed on the following 10 out of the 39 genes: neuroendocrine-associated genes (INSM1, ASCL1, NRCAM, and SNAP25), one gene centered in the gene regulatory network (NUF2), and 5 possible cancer-associated genes (PTP4A3, RFC4, REST, APEH, and FBLN2).
Caption: Figure 2: The mRNA level of INSM1, ASCII, NRCAM, SNAP25, NUF2, PTP4A3, RFC4, REST, APEH, and FBLN2 in SCEC.
Another research group found that miR-1 inhibits fibulin-2 (Fbln2
) expression and thereby it abolishes activation of TGF[beta] signaling and extracellular matrix remodeling in hypertrophic heart [11,12].
Recientemente se han encontrado nuevos genes metilados relacionados con cancer de mama, EMELIN 2, (18p11.3), SALL 1 (16q12.1), DBCI (9q32-q33), FBLN2
(3p25.1), CIDE, (18p11.21) (41).
We performed quantitative real-time PCR (q-RT-PCR) using random hexamer primers and Taqman gene expression assays (Applied Biosystems Prism 7700) for von Willebrand factor (VWF)12 (Hs01109460_m1); leptin (LEP) (Hs00174877_m1); matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase) (MMP2) (Hs00234422_m1); fibulin 1 (FBLN1) (Hs00242546_m1); and fibulin 2 (FBLN2) (Hs01002054_m1).
q-RT-PCR analysis of 5 of the 11 most upregulated genes (FBLN1, VWF, FBLN2, MMP2, and LEP) showed that 3 genes, FBLN1, VWF, and FBLN2, were significantly upregulated (Table 2).
 Human genes: VWF, von Willebrand factor; LEP, leptin; MMP2, matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase); FBLN1, fibulin 1; FBLN2, fibulin 2; ACTS, actin, beta; CRISPLD2, Cysteine-rich secretory protein LCCL domain containing 2; SERPINF1, Serpin peptidase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1; ITM2A, Integral membrane protein 2A; LEPR, leptin receptor; ZFP36L2, zinc finger protein 36, C3H type-like 2; ELN, elastin.
Developmental genes such as Bcan, Fbln2
, Itih3, Ncam1, Tnr, and Vcan modulate NSC differentiation via Wnt/[beta]-catenin pathway at early stage of differentiation and TGF-[beta] signaling pathway at later (7 day) stage.
ENSMUSG00000086859 targeted genes like Efna1, Fbln1, and Fbln2. ENSMUSG00000066057 regulated the DEGs, such as CYP51, FDPS, and Eif2s1 (Figure 2).
A series of positively regulated target genes were significantly enriched in the pathways of cholesterol biosynthesis (e.g., Cyp51 and Fdps) and elastic fibre formation (e.g., Fbln1, Fbln2, and Fbln5) (Table 2).
The DEGs positively regulated by DE-lncRNAs were mainly enriched in the functions about the development of blood vessel (e.g., Efna1 and Efnb2), as well as the pathways of cholesterol biosynthesis (e.g., Cyp51 and Fdps) and elastic fibre formation (e.g., Fbln1, Fbln2, and Fbln5).
In this study, several other DEGs positively regulated by DE-lncRNAs (e.g., Fbln1, Fbln2, and Fbln5) were significantly enriched in the pathway of elastic fibre formation.