Because epigenetic alterations play an important role in prostate carcinogenesis and owing to the relative shortage of validated data on miRNA altered expression in PCa, we aimed to identify and validate miRNAs upregulated in this malignancy.
With reference to the original 2D DiGE analysis, of the 58 upregulated spots that yielded protein identifications, 34 (59%) contained at least one protein whose corresponding mRNA was validated as being upregulated by knockdown of UPF1, and in 26 cases (45%) the upregulated mRNA corresponded to the major protein component of the spot, as indicated by MASCOT scores (see Additional file 2).
TPM3, CAMP, ERp29, and HSPA8 were validated as being upregulated in three CRC cases with a 1.68, 1.88, 2.14, and 2.44-fold change, respectively).
Using qPCR, we validated that XM_005184828 is upregulated in Y M testis (P < 10 − 4 ) and XM_005176585 is upregulated in III M testis (P < 0.01) (supplementary fig. S6 and supplementary data, Supplementary Material online).
Adipose differentiation-related protein (ADFP, BC005127), which is a marker of adipocyte differentiation and lipid storage [5], was significantly upregulated and validated by IHC (Fig. 4b).
Doxpr transcript was validated to be significantly upregulated in T2Bo_A using a different BR sample pair from those used to generate the microarray data.
We further extended our studies to triple-negative and Herceptin-resistant breast cancer phenotype, where EGFR expression is upregulated, and validated the importance of AnxA2 in keeping EGFR signalling in an activated state, leading to cancer cell proliferation and migration.
Experimentally validated target genes for microRNAs upregulated in gastric cancer.
miR-25 (P=0.0030) and miR-19b (P=0.0020) were validated to be more significantly upregulated in ESCC patients than in healthy volunteers.
Among the genes that are differentially regulated between WT and CD38-KO mice (Table S1), we validated 4 genes which were upregulated in the mutant mice (Fig. 3B).
