human fetal lung fibroblast hlf1  (Procell Inc)

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