Journal: MedComm

Article Title: Sphingosine‐1‐Phosphate Receptor 3 Confers Tumor Metastasis in Lung Cancer Resistant to Third‐Generation EGFR Inhibitor

doi: 10.1002/mco2.70744

Figure Lengend Snippet: S1PR3 is a key mediator of metastasis in EGFR inhibitor‐resistant lung cancer cells. (A) Migration or (B) invasion of parental cells (NCI‐H1975) and resistant cells (120067R and AZD9291R) detected by trans‐well assays. (C) GO enrichment analysis of differentially expressed genes (fold change ≥ 2) between parental and resistant cells as identified through RNA‐seq analysis. (D) Gene or (E) protein expression of S1PR3 in parental cells and resistant cells was analyzed by RT‐PCR assay or immunoblotting. (F) Migration and invasion of 120067R cells after siS1PR3 transfection or (G) treated with S1PR3 inhibitor TY‐52156. (H) Migration and invasion of NCI‐H1975 cells exogenous overexpressed S1PR3. (I) IHC staining of S1PR3 on human tissue specimens from adjacent normal tissues, lung cancer, and AZD9291‐resistant lung cancer. (J) Overall survival for lung adenocarcinoma patients with high or low S1PR3 expression was analyzed by Kaplan–Meier survival analysis. Scale bar, 100 µm. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: NCI‐H1975 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, RNA Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Immunohistochemistry

Journal: MedComm

Article Title: Sphingosine‐1‐Phosphate Receptor 3 Confers Tumor Metastasis in Lung Cancer Resistant to Third‐Generation EGFR Inhibitor

doi: 10.1002/mco2.70744

Figure Lengend Snippet: S1PR3 contributed to the metastasis in EGFR TKIs‐resistant cells through the RAC1‐PAK1 signaling pathway. (A) Schematic diagram of S1PR3 downstream pathways. (B) The protein expression of NCI‐H1975 cells and 120067R cells was detected by immunoblotting. (C) Immunoblotting on 120067R cells after S1PR3 knockdown or (D) treated with S1PR3 inhibitor TY‐52156. (E) Immunoblotting of NCI‐H1975 cells showed exogenous overexpression of S1PR3. (F) Migration and invasion of 120067R cells after RAC1 or (G) PAK1 knockdown. (H) Migration and invasion or (I) immunoblotting of 120067R cells showed exogenous overexpression of RAC1 and subsequently treated with S1PR3 inhibitor. Scale bar, 100 µm. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: NCI‐H1975 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Migration

Journal: MedComm

Article Title: Sphingosine‐1‐Phosphate Receptor 3 Confers Tumor Metastasis in Lung Cancer Resistant to Third‐Generation EGFR Inhibitor

doi: 10.1002/mco2.70744

Figure Lengend Snippet: S1PR3‐RAC1‐PAK1 axis is associated with the EMT progression of resistant cells. (A) Gene or (B) protein expression of EMT markers detected by RT‐PCR assays and immunoblotting. (C) Transcriptome signature of EMT in S1PR3 high group (S1PR3‐H) and S1PR3 low group (S1PR3‐L) of lung adenocarcinoma patients. (D) The association between S1PR3 and EMT‐related genes level in lung adenocarcinoma patients. (E) Immunoblotting on 120067R cells after S1PR3 knockdown or (F) treatment with S1PR3 inhibitor. (G) Immunoblotting of NCI‐H1975 cells showed exogenous overexpression of S1PR3. (H) Immunoblotting on 120067R cells after PAK1 knockdown or (I) treatment with PAK1 inhibitor. (J) Immunoblotting data of NCI‐H1975 cells showed exogenous overexpression of PAK1. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: NCI‐H1975 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Knockdown, Over Expression