Journal: Neoplasia (New York, N.Y.)

Article Title: Estrogen/GPR30 Signaling Contributes to the Malignant Potentials of ER-Negative Cervical Adenocarcinoma via Regulation of Claudin-1 Expression

doi: 10.1016/j.neo.2018.08.010

Figure Lengend Snippet: Estrogen induces claudin-1 expression in cervical adenocarcinoma cells. (A) CLDN1 expression was induced by E2 treatment (24 hours) in CAC-1, Hela229, and HCA1 cells. (B) Immunofluorescence of CLDN1 (green) was induced by E2 treatment (10 nM, 24 hours) in HCA1 and Hela229 cells. (C) Expression profile of GPR30 (membrane-bound estrogen receptor) and ERα (classical estrogen receptor) in cervical adenocarcinoma cells. MCF7 cells (breast cancer cells) were used as a positive control for ERα expression. Western blotting. (D) Immunofluorescence of GPR30 (green) in cervical adenocarcinoma cells. (E) CLDN1 expression was induced by G1 (GPR30 selective agonist), but not by PPT (ERα selective agonist) or DPN (ERβ selective agonist), treatment (10 nM, 24 hours) in cervical adenocarcinoma cells. The relative intensity of each band is shown over each immunoblot after normalization for the levels of actin. CLDN1: claudin-1.

Article Snippet: The human cervical adenocarcinoma cell lines Hela229, and OMC4 were purchased from RIKEN Bio-Resource Center (Tsukuba, Japan), and HCA1 was from JCRB cell bank (Osaka, Japan).

Techniques: Expressing, Immunofluorescence, Membrane, Positive Control, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: Estrogen/GPR30 Signaling Contributes to the Malignant Potentials of ER-Negative Cervical Adenocarcinoma via Regulation of Claudin-1 Expression

doi: 10.1016/j.neo.2018.08.010

Figure Lengend Snippet: GPR30 mediates induction of claudin-1 expression in cervical adenocarcinoma cells. (A) CLDN1 expression was induced by G1 treatment (24 hours) in CAC-1, Hela229, and HCA1 cells. (B) Immunofluorescence of CLDN1 was induced by G1 treatment (10 nM, 24 hours) in Hela229 and HCA1 cells. (C) G1-induced CLDN1 expression was inhibited by pretreatment with G15 (GPR30 selective antagonist) for 24 hours in CAC-1 and Hela229 cells. (D) GPR30 expression was knocked down by GPR30-specific siRNA in HCA1 cells. (E) G1-induced CLDN1 expression was inhibited by knockdown of GPR30 expression with GRP30-specific siRNA in HCA1 cells. (F) G1 treatment (10 nM) increased phosphorylation of ERK and Akt in Hela229 cells. (G) G1-induced CLDN1 expression was inhibited by pretreatment with U0126 (10 μM) and LY294002 (10 μM) in CAC-1 and Hela229 cells. (A, C-G) Western blotting and (B) confocal microscopy. The relative intensity of each band is shown over each immunoblot after normalization for the levels of actin. CLDN1: claudin-1.

Article Snippet: The human cervical adenocarcinoma cell lines Hela229, and OMC4 were purchased from RIKEN Bio-Resource Center (Tsukuba, Japan), and HCA1 was from JCRB cell bank (Osaka, Japan).

Techniques: Expressing, Immunofluorescence, Knockdown, Phospho-proteomics, Western Blot, Confocal Microscopy