Relative mRNA levels were calculated with a delta-delta-Ct method normalized to 18S rRNA levels as an internal control using the following primers: TaqMan Gene Expression Assay for Mouse Map2k1
(Applied Biosystems, Carlsbad, CA, USA; Assay ID: Mm00435940_m1) and Predeveloped TaqMan Assay Reagents 18S rRNA (20x) (Applied Biosystems, Catalog Number 4319413E).
We suppose that components of the MAPK- and PI3K/mTOR/PTEN-signalling cascades (FOS, MAPK7, MAPK13, MAP2K1
, MTOR, PI3CA, PTEN, RHEB, RPS6KB1, and TP53) play an important role in cell motility [35, 36] and represent functional modules of this interactive network.
To explore whether the patient would benefit from the BRAF inhibitor therapy, BRAF and MAP2K1
mutations were further tested.
In the present study, miR-195 potentially targeted several genes that are involved in multiple signaling pathways, i.e., the Gadd45g, Map2k1
, Mras, and Raf1 genes, which are involved in the MAPK signaling pathway.
Tested genes include AKT1, ALK, AXL, BRAF, CALCA, CCND1, CTNNB1, DDR2, EGFR, ERBB2, FGFR1, FGFR2, FGFR3, GNAS, HRAS, IDH1, IDH2, KRAS, KRT20, KRT7, MAP2K1
, MET, NRAS, NRG1, NTRK1, NTRK2, NTRK3, PIK3CA, PPARG, PTH, RAF1, RET, ROS1, SLC5A5, THADA, TTF1, CHMP2A, GPI, RAB7A, and VCP.
Samples With Previously Identified Heterozygous Variants That Were Used for Validation of the Combined Disease Panels Gene Transcript Variant Type CDKL5 NM_001037343.1 c.2384A>C, SNV p.Asn795Thr CDKL5 NM_001037343.1 c.380A>G, SNV p.His127Arg FGFR1 NM_023110.2 c.755C>G, SNV p.Pro252Arg FGFR2 NM_000141.4 c.1018T>C, SNV p.Tyr340His FGFR3 NM_001163213.1 c.749C>G, SNV p.Pro250Arg FLCN NM_144997.5 c.1285delC, Indel p.His429ThrfsX39 FLCN NM_144997.5 c.1285dupC, Indel p.His429ProfsX27 GJB2 NM_004004.5 c.35delG, Indel p.Gly12ValfsX2 MAP2K1
NM_002755.3 c.388T>C, SNV p.Tyr130His MYO7A NM_000260.3 c.3719G>A, SNV p.
They enrolled lung adenocarcinoma patients with a [less than or equal to] 15 pack-year smoking history whose tumors previously tested "negative" for alterations in 11 genes (mutations in EGFR, ERBB2, KRAS, NRAS, BRAF, MAP2K1
, PIK3CA, and AKT1, and fusions involving ALK, ROS1, and RET).
(22) Mutations in MAP2K1
, which encodes the dual-specificity kinase MEK1 protein in the MAPK pathway, were subsequently identified in 27.5% of LCH cases, thus explaining MAPK pathway activation in the absence of BRAF mutation.
We established a deep-sequencing assay on the GS Junior platform with 454 GS Junior Titanium chemistry (Roche Applied Science, Indianapolis, Indiana) for simultaneous, prospective, semiquantitative mutational assessment of the 10 most commonly mutated genes in lung adenocarcinoma: KRAS, EGFR, ERBB2 (HER2), BRAF, NRAS, HRAS (Harvey rat sarcoma viral oncogene homolog), PIK3CA, MAP2K1
(MEK1), PTEN (phosphatase and tensin homolog), and AKT1 covering all genes analyzed for mutation by the Lung Cancer Mutation Consortium, with the exception of the EML4-ALK translocation and MET amplifications.
However, downstream mutations of MAP2K1
(MEK1) have been identified in approximately one-third of HCL-vs and in most IGHV4-34-expressing classic HCLs.
Eight percent of melanomas showed gain-of-function mutations in the MAP2K1
(encoding MEK1) and MAP2K2 (encoding MEK2) genes, (476) resulting in constitutive activation of downstream ERK MAPK.
Other alterations occur at lower frequencies in NSCLC, including mutations in AKT1, (24) BRAF, (24) and MAP2K1
, (25) and amplifications and mutations in MET (26) (for a review see Pao and Girard (27)).