Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Itraconazole induces apoptosis and cell cycle arrest via inhibiting Hedgehog signaling in gastric cancer cells

doi: 10.1186/s13046-017-0526-0

Figure Lengend Snippet: Itraconazole inhibits the growth of gastric cancer cells in vitro and in vivo. a Decrease in viability of gastric cancer cells (AGS and MKN45) following treatment with increasing concentrations of itraconazole. Cell viability was measured by the CCK-8 assay. Data represent the mean of three experiments (mean ± SD). * p < 0.05; ** P <0.01 vs DMSO-treated cells. ITRA, itraconazole. b Repression of colony formation of gastric cancer cells following treatment with different concentrations of itraconazole. AGS and MKN45 cells, treated with itraconazole, were grown in a 6-well plate and colonies were scored after 14 days. Cell clusters containing more than 50 cells under a microscope were considered as colonies. Histograms show the formation rate of colonies. Data represent the mean of three experiments (mean ± SD). * P <0.05 and ** P <0.01, vs DMSO treated cells. c Itraconazole enhances the inhibitory effect of 5-FU on gastric cancer cells. AGS and MKN45 cells were treated with 10 μM itraconazole, 10 μM 5-FU or both. Data represent the average of three experiments (mean ± SD). * P <0.05 and ** P <0.01 vs DMSO-treated cells. d Itraconazole inhibits growth of gastric cancer xenografts. Nude mice with AGS subcutaneous tumor xenografts were treated with vehicle ( n = 8), itraconazole 75 mg/kg twice daily by oral administration (ITRA; n = 8), 5-FU 20 mg/kg every 2 days by intraperitoneal injection ( n = 8), or a combination of itraconazole and 5-FU (ITRA + 5-FU; n = 6). Nude mice with MKN45 subcutaneous tumor xenografts were treated with vehicle ( n = 10), itraconazole 75 mg/kg twice daily by oral administration, 5-FU 20 mg/kg every 2 days by intraperitoneal injection ( n = 8), or a combination of itraconazole and 5-FU (ITRA + 5-FU; n = 8). Mean ± SD tumor volumes are reported for each treatment group

Article Snippet: Two gastric cancer cell lines, MKN45 (KCLB 80103) and AGS (ATCC CRL 1739), were maintained in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (ScienCell) and 1% penicillin-streptomycin sulfate (Gibco).

Techniques: In Vitro, In Vivo, CCK-8 Assay, Microscopy, Injection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Itraconazole induces apoptosis and cell cycle arrest via inhibiting Hedgehog signaling in gastric cancer cells

doi: 10.1186/s13046-017-0526-0

Figure Lengend Snippet: Effects of itraconazole on cell cycle distribution and apoptosis of gastric cancer cells. a Cell cycle distributions were detected by flow cytometry analysis in AGS and MKN45 cells after treatment with different concentrations of itraconazole or DMSO for 72 h. Data represent the mean of three experiments (mean ± SD). * p < 0.05; ** P <0.01, compared to DMSO treated cells. b The expression of cell cycle-related proteins is examined by immunoblot assay. AGS and MKN45 cells are harvested after itraconazole treatment for 48 h. GAPDH is used as a loading control. c Cell apoptosis is determined by flow cytometry analysis in AGS and MKN45 cells after treatment with different concentrations of itraconazole or DMSO for 72 h. Data represent the average of three experiments (mean ± SD). * P <0.05 and ** P <0.01 vs DMSO treated cells. d The Bax expression at protein level is examined by immunoblot assay. GAPDH is used as a loading control

Article Snippet: Two gastric cancer cell lines, MKN45 (KCLB 80103) and AGS (ATCC CRL 1739), were maintained in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (ScienCell) and 1% penicillin-streptomycin sulfate (Gibco).

Techniques: Flow Cytometry, Expressing, Western Blot, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Itraconazole induces apoptosis and cell cycle arrest via inhibiting Hedgehog signaling in gastric cancer cells

doi: 10.1186/s13046-017-0526-0

Figure Lengend Snippet: Itraconazole inhibits the expression of Gli1 in gastric cancer cells. a Expressions of Shh, Ptch, Smo, and Gli1 in MKN45 and AGS cells treated with increasing concentrations of itraconazole for 48 h. Data represent the mean ± SD of 3 determinations and GAPDH is used as the internal control. * P < 0.05; ** p < 0.01. b MKN45 and AGS cells are treated with DMSO or indicated concentrations of itraconazole for 48 h, and expressions of major molecules of Hh signaling are determined by immunoblot. A representative blot from three independent experiments is shown. c MKN45 and AGS cells are transiently transfected with Gli1-pGL3, Smo -pGL3 or pGL3-Basic vector and pRL-TK, followed by treatment with 10 μM itraconazole or DMSO. After 48 h, Gli1-pGL3, Smo -pGL3 or pGL3-Basic vector reporter activities were measured and normalized to Renilla luciferase activity. Data represent the mean of three experiments (mean ± SD). * p < 0.05 and ** P <0.01 vs DMSO treated cells

Article Snippet: Two gastric cancer cell lines, MKN45 (KCLB 80103) and AGS (ATCC CRL 1739), were maintained in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (ScienCell) and 1% penicillin-streptomycin sulfate (Gibco).

Techniques: Expressing, Control, Western Blot, Transfection, Plasmid Preparation, Luciferase, Activity Assay