Journal: Oncology Reports

Article Title: Aprepitant inhibits the progression of esophageal squamous cancer by blocking the truncated neurokinin‑1 receptor

doi: 10.3892/or.2023.8568

Figure Lengend Snippet: Expression patterns of NK1R in human ESCC cell lines and human tumor specimens. (A) mRNA expression of NK1R is shown as fold expression relative to human FB. (B) Western blot analysis for specific antibodies binding to either fl-NK1R or both tr- and fl-NK1R. (C and D) mRNA expression of (C) tr-NK1R and (D) fl-NK1R in normal esophageal and ESCC specimens. (E) Relative mRNA expression of tr-NK1R and fl-NK1R in ESCC. (F) H&E staining from human ESCC tissue and IHC staining for NK1R and SP are shown at higher magnification for both tumor and normal tissues. (G) CD163 and SP expression in ESCC tissues detected by immunofluorescence. Green: CD163; Red: SP; Blue: DAPI. (H) Cells were stained with antibodies against CD68 and CD163. Representative IHC images of ESCC and normal tissues with statistics of the corresponding expression levels are shown. (I) Microscopic images showing the differentiated progression from M0 to M2 and expression of M2 markers in THP-1 cells treated with IL-4 and IL-13. (J) SP secretion from FB, M2 macrophage and ESCC cells detected by ELISA. Scale bars, 100 µm. *P<0.05. NK1R, neurokinin-1 receptor; ESCC, esophageal squamous cell carcinoma; FB, fibroblasts; fl, full length; tr, truncated; SP, substance P; IHC, immunohistochemical.

Article Snippet: Human fibroblasts (Human fetal lung fibroblast, HLF1, cat. no. CL-0106) were purchased from Procell Life Science & Technology Co., Ltd., and cultured in Ham's F-12K (Gibco; Thermo Fisher Scientific, Inc.) containing 10% fetal calf serum and 1% penicillin/streptomycin.

Techniques: Expressing, Western Blot, Binding Assay, Staining, Immunohistochemistry, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: Oncology Reports

Article Title: Aprepitant inhibits the progression of esophageal squamous cancer by blocking the truncated neurokinin‑1 receptor

doi: 10.3892/or.2023.8568

Figure Lengend Snippet: AP inhibits the SP-induced proliferation of human ESCC cell lines. (A) TE1, KYSE-150, and KYSE-170 cells were stimulated with different concentrations of SP to clarify its mitogenic potential in ESCC cells. (B) CCK-8 assays determining cell survival after treatment with aprepitant for 48 h are shown for the cell lines TE1, KYSE-150 and KYSE-170, and for human fibroblasts. Based on these data, IC 50 (µM) was calculated and compared with fibroblasts for statistical analysis. (C) TE1, KYSE-150 and KYSE-170 cells were treated with anti-SP, anti-NK1R and isotype antibodies at a final concentration of 1:100. The effects were analyzed with CCK-8 proliferation assays. (D) All three ESCC cell lines were treated with different concentrations of SP and 25 µM aprepitant, and survival effects were detected by CCK-8 assay. (E) The knockdown efficiency of fl-NK1R and tr-NK1R detected by western blot analysis in TE1, KYSE-150 and KYSE-170 cells. (F) The proliferation of TE1, KYSE-150 and KYSE170 cells, respectively, after fl-NK1R and tr-NK1R were knocked down. (G) After fl-NK1R and tr-NK1R were knocked down respectively in TE1, KYSE-150 and KYSE-170 cells, IC 50 of aprepitant was applied to observe the inhibitory rate. *P<0.05 and **P<0.01. AP, aprepitant; SP, substance P; ESCC, esophageal squamous cell carcinoma; CCK-8, Cell Counting Kit-8; IC 50 , half maximal inhibitory concentration; NK1R, neurokinin-1 receptor; fl, full length; tr, truncated; si-, small interfering; NC, negative control.

Article Snippet: Human fibroblasts (Human fetal lung fibroblast, HLF1, cat. no. CL-0106) were purchased from Procell Life Science & Technology Co., Ltd., and cultured in Ham's F-12K (Gibco; Thermo Fisher Scientific, Inc.) containing 10% fetal calf serum and 1% penicillin/streptomycin.

Techniques: CCK-8 Assay, Concentration Assay, Knockdown, Western Blot, Cell Counting, Negative Control

Journal: BMC Genomics

Article Title: Distribution of segmental duplications in the context of higher order chromatin organisation of human chromosome 7

doi: 10.1186/1471-2164-15-537

Figure Lengend Snippet: Distribution of segmental duplications (SDs) and bundled long distance interactions in relation to acetylation of H4K8, transcriptional activity and lamina associated domains on human chromosome 7 (derived from IMR90 unless indicated otherwise). A) H4K8 acetylation profile, dark yellow: hyperacetylation of H4K8; blue: hypoacetylation of H4K8. B) the red and blue curve represent RNA-seq read counts/100 kb bin for coding and non-coding RNA, respectively (IMR91L). C) grey areas underlying the two histograms mark lamina associated domains (LADs, Tig3 cells). D) idiogram of chromosome 7, the Williams-Beuren syndrome region is highlighted in yellow beside the idiogram (at 72-74 Mb, hg18). E) transparent blue shading of the idiogram illustrates the inversion-affected segments of chromosome 7 depicted in Figure A-C. Bundled long distance interactions (F) and segmental duplications (G) are depicted in the inner circle; green ribbons: long distance interactions between genomic regions; grey: SDs with sequence similarity <98%; yellow: SDs with sequence similarity 98-99%; orange: SDs with sequence similarity >99%.

Article Snippet: Human fetal lung fibroblast cell lines IMR91L (male) and IMR90 (female) were obtained from the Coriell Institute for Medical Research.

Techniques: Activity Assay, Derivative Assay, RNA Sequencing, Sequencing

Journal: BMC Genomics

Article Title: Distribution of segmental duplications in the context of higher order chromatin organisation of human chromosome 7

doi: 10.1186/1471-2164-15-537

Figure Lengend Snippet: Higher order chromatin organisation and SD localisation around the Williams-Beuren syndrome region. All data are referring to genome release hg19 and are derived from IMR90 unless indicated otherwise. The proximal, central and distal SD clusters (P, C, D) of the 7q11 segment encompassing 4.8 Mb are highlighted in green within the chromosome banding track. A-C) localisation of SDs; colouring according to sequence similarity; grey: <98%, yellow: 98%-99%; orange: >99%; D) genomic interval commonly deleted in WBS and the distal 7q11.23 deletion syndrome; E) topological domains as defined by Dixon et al. ; F) topological domains identified in the corresponding region in mouse after conversion to human hg19. Note that the murine topological domain homologous to sequences deleted in the distal 7q11.23 syndrome is not fully represented due to a break of synteny within this genomic interval. See Figure for details; G-H) heatmap and arc view of CTCF binding sites as detected by ChIA-PET in MCF7; I) number of G4 motifs/100 kb bins; J) average GC-content within 100 kb bins; K) number of Alu repeats/100 kb bin; L) number of structural variants as annotated by Database of Genomic Variance (DGV) , *maximum of 1080 CNVs not shown; M) log2 ratio scores of the LaminB1 DamID Map (Tig3 cells) as reported by Guelen et al. ; N) log2 ratio scores of DNA regions prone to early apoptotic DNA degradation in 20 kb windows, turquoise: degraded DNA segments; O) log2 ratio scores of H4K8 acetylation profile in 20 kb windows, blue: hyperacetylation, grey: hypoacetylation; P) red curve representing the sum of all intrachromosomal interaction counts/bin divided by the median number of interactions for all bins of chromosome 7; Q) percentage of interactions categorised according to their interaction span size ; light grey: <0.5 Mb, grey: 0.5-1 Mb, light blue: 1–5 Mb, light brown: 5–10 Mb, dark grey: 10–25 Mb, black: ≥25 Mb. Gaps in this plot are due to alignment problems of Hi-C data in regions harbouring SDs with high sequence similarity.

Article Snippet: Human fetal lung fibroblast cell lines IMR91L (male) and IMR90 (female) were obtained from the Coriell Institute for Medical Research.

Techniques: Derivative Assay, Sequencing, Binding Assay, ChIA Pet Assay, Hi-C

Journal: BMC Genomics

Article Title: Distribution of segmental duplications in the context of higher order chromatin organisation of human chromosome 7

doi: 10.1186/1471-2164-15-537

Figure Lengend Snippet: Cross-species comparison showing that SDs next to the WBS locus have inserted at topological domain borders. Hi-C interactions and topological domains in the human fetal fibroblast cell line IMR90 are shown in dark green in the upper part as triangle view and bars, respectively. SDs with sequence similarity of 98%-99% and above 99%, respectively, (shown in yellow and orange in the SDs track) coincide with gaps within the Hi-C data. SD distribution and Hi-C data of the corresponding region in mouse are given in the lower part of the image. The position of FKBP6 and WBSCR16 , the human orthologues of the two genes next to the murine topological domain borders are highlighted in green and red, respectively. The intervals commonly affected in WBS and the distal 7q11.23 syndrome are indicated by pale red bars. Note that the region distal to SRRM3 including the distal SD block are homologous to a different mouse chromosome.

Article Snippet: Human fetal lung fibroblast cell lines IMR91L (male) and IMR90 (female) were obtained from the Coriell Institute for Medical Research.

Techniques: Comparison, Hi-C, Sequencing, Blocking Assay