The reference gene ywhaz
encoding phospholipase A2 was used for these experiments (18).
Multiple miRNA target prediction algorithms indicated the well-studied oncogene YWHAZ
as a direct target of miR-375, which was further confirmed by the luciferase reporter assay, qPCR, and Western blotting.
Targets Primers Amplicon (bp) genes 1 LPL F-5'aatgaagagatgaacggaacg-3' 119 R-5'gcactttccaaccaggatgt-3' 2 SCD F- 5'cccagctgtcagagaaaagg- 3' 115 R- 5'gatgaagcacaacagcagga- 3' 3 SREBF1 F-5'ctgctatgcaggcagcac- 3' 99 R- 5'ggttgatgggcagcttgt- 3' 4 YWHAZ
F-5'tgtaggagcccgtaggtcatct-3' 102 R-5'ttctctctgtattctcgagccatct-3 5 PRKAA2 F-5'accctcccatttgatgatga-3' 97 R-5'tggcaacagaacgattgaga-3' Gene No.
Ct values of genes of interests were normalised against the averaged Ct values of two housekeeping genes (succinate dehydrogenase complex flavoprotein subunit A (SDHA) and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ
A set of three genes was used for data normalization, namely Hypoxanthine Phosphoribosyltransferase 1 (HPRT), Importine 8 (IPO8) and tyrosine 3-mono-oxygenase/tryptophan 5-monooxygenase activation protein, zeta (YWHAZ
Six potential internal control genes (ICG) were tested [i.e., glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein S9 (RPS9), ubiquitously expressed transcript (UXT), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (YWHAZ
), glucose-6-phosphate dehydrogenase (G6PD), and mitochondrial ribosomal protein L39 (MRPL39)] based on prior publications [40, 41] using geNorm .
Ruenauver et al., "Somatic copy number alterations by whole-exome sequencing implicates YWHAZ
and PTK2 in castration-resistant prostate cancer," The Journal of Pathology, vol.
Then, the median mutation loads for each cancer type and the median NEIL1, NEIL2, and NEIL3 expression values normalized to the expression value of the constitutive housekeeping gene YWHAZ
(ENSG00000164924)  were analyzed to identify correlations.
defined tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta (YWHAZ
), as a target of miR-451 and overexpression of miR-451 caused reduction of p38-MAPK signaling via suppression of YWHAZ
, revealing the potential therapeutic role of miR-451 since p38-MAPK pathway was positively involved in DN .
Genes such as MCM7 (4.78), DFFA (5.24), and PRPF38A (4.28) are engaged in gene regulation while E2F5 (6.60), RPS6KA3 (4.77), and YWHAZ
(5.18) are associated with cell proliferation.
Amplification of LAPTM4B and YWHAZ
contributes to chemotherapy resistance and recurrence of breast cancer.
Three reference genes were selected for normalization (MoDC: TATA-binding protein [TBP], tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta [YWHAZ], and beta-2-microglobulin [B2M]; IPEC-J2: TBP, YWHAZ
, and glyceraldehyde-3phosphate dehydrogenase [GAPDH]).