Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 was upregulated in oral squamous cell carcinoma (OSCC) tissue and associated with clinical disease stages in human OSCC. (A) Correlation CX3CL1 gene expression with the normal and tumour cells using GEO microarray 3524 OSCC tissue samples. (B) CX3CL1 expression profiles in 497 OSCC tissue specimens were analysed obtained from the Cancer Genome Atlas (TCGA) database. (C,D) OSCC specimens were subjected to IHC staining. (E) Kaplan–Meier survival analysis of the associations between high or low plasma levels of CX3CL1 expression and overall survival of OSCC patients. * p < 0.05 compared with controls.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Gene Expression, Microarray, Expressing, Immunohistochemistry, Clinical Proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 upregulates human oral squamous cell carcinoma (OSCC) cell migration and invasion. (A‐C) OSCC cells were incubated with different concentrations of CX3CL1 for 24 h; then, cell migration was assessed using the (A) in vitro wound‐healing assay, (B,C) the Transwell assay. (D) Quantified result of cell migration with CX3CL1 neutralizing antibody. (E) Quantified result of cell invasion with CX3CL1 neutralizing antibody. Results are expressed as the mean ± SD of four independent experiments. * p < 0.05 as compared with controls.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Migration, Incubation, In Vitro, Wound Healing Assay, Transwell Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 activates tumour cell migration via the ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A) ICAM‐1 expression in the normal and tumour cells obtained from the TCGA dataset analysis. (B) VCAM‐1 expression in the normal and tumour cells obtained from the TCGA dataset analysis. (C) Correlation analysis of CX3CL1 and ICAM‐1 expression using the TIMER2.0 database. (D) Correlation analysis of CX3CL1 and VCAM‐1 expression using the TIMER2.0 database. (E) Quantified result of ICAM‐1 gene expression with CX3CL1 recombinant protein (30 ng/mL) treatment. (F) Quantified result of VCAM‐1 gene expression with CX3CL1 recombinant protein (30 ng/mL) treatment. (G) Protein expression of ICAM‐1 with concentration‐depended CX3CL1 treatment. (H) Protein expression of VCAM‐1 with concentration‐depended CX3CL1 treatment. (I) Quantified result of cell migration with ICAM‐1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (J) Quantified result of cell migration with ICAM‐1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Migration, Expressing, Gene Expression, Recombinant, Concentration Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 upregulates cell motility and ICAM‐1 expression via its receptor CX3CR1. (A) Levels of CX3CR1 mRNA expression in normal tongue tissue and human oral squamous cell carcinoma (OSCC) tumour tissue were analysed using records from the GEO data set GSE13601. (B) Levels of CX3CR1 mRNA expression in different N stages of OSCC tumour tissue were analysed using records from the GEO data set GSE78060. (C) Quantified result of cell movement with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (D) Quantified result of cell migration with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Quantified resulting of ICAM‐1 expression with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (F) Quantified result of cell movement with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (G) Quantified result of cell migration with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (H) Quantified result of ICAM‐1 expression with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Expressing, Recombinant, Migration, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 triggers the phosphorylation of PLCβ/PKCα/c‐Src to signal the cell movement, migration and ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A‐C) Quantified result of cell movement, cell migration and ICAM‐1 expression with signalling inhibitors and CX3CL1 recombinant protein (30 ng/mL) treatment. (D‐E) Protein expression of phosphorylated PLCβ, PKCα and c‐Src with CX3CL1 recombinant protein (30 ng/mL) treatment in SCC4 cell and SAS cell. (F‐H) Quantified result of cell movement, cell migration and ICAM‐1 expression with PLCβ, PKCα or c‐Src siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Phospho-proteomics, Migration, Expressing, Recombinant, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 promotes the phosphorylation of c‐Jun to upregulate the cell motility and ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A) Quantified result of CX3CL1‐induced cell movement with c‐Jun inhibitors (B) Quantified result of CX3CL1‐induced cell migration with c‐Jun inhibitors (C) Quantified result of CX3CL1‐induced ICAM‐1 expression with c‐Jun inhibitors (D) Protein expression of phosphorylated c‐Jun with CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Quantified result of CX3CL1‐induced cell movement with c‐Jun siRNA treatment. (F) Quantified result of CX3CL1‐induced cell migration with c‐Jun siRNA treatment. (G) Quantified result of CX3CL1‐induced ICAM‐1 expression with c‐Jun siRNA treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Phospho-proteomics, Expressing, Migration, Recombinant, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 promotes the translocation of c‐Jun and promoter binding of AP‐1 formation in human oral squamous cell carcinoma (OSCC) cells. (A) Immunofluorescent staining showed that c‐Jun translocated into the nucleus after cell were treated with CX3CL1; nuclear translocation was prevented when the cells were pre‐treated with CX3CR1 mAb, U73122, GF109203 or PP2. (B) Quantified analysis of c‐Jun activation with CX3CL1 using Western blot. (C) CX3CL1 treatment upregulated AP‐1 luciferase activity. (D) Quantified analysis of AP‐1 luciferase activity with specific inhibitors and CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Chromatin immunoprecipitation analysis of c‐Jun and AP‐1 promoter binding site with neutralizing antibody and specific inhibitors. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Three human OSCC cell lines (SCC4, SCC25 and SAS cells) were acquired from the Bioresource Collection and Research Center (BCRC).

Techniques: Translocation Assay, Binding Assay, Staining, Activation Assay, Western Blot, Luciferase, Activity Assay, Recombinant, Chromatin Immunoprecipitation, Control

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Rapamycin enhances the anti-angiogenesis and anti-proliferation ability of YM155 in oral squamous cell carcinoma.

doi: 10.1177/1010428317706213

Figure Lengend Snippet: Figure 1. The inhibitory effect of rapamycin and YM155 on the proliferation ability of OSCC cell lines. (a) The cell growth curve of the four OSCC cell lines (SCC9, SCC15, SCC25, and CAL27) treated with varied rapamycin doses. (b) The cell growth curve of SCC9 cell lines treated with different doses of YM155 and rapamycin. (c) The cell growth curve of SCC15 cell lines treated with different doses of YM155 and rapamycin. (d) The cell growth curve of SCC25 cell lines treated with different doses of YM155 and rapamycin. (e) The cell growth curve of CAL27 cell lines treated with different doses of YM155 and rapamycin. All data are presented as mean ± SD for three different experiments performed in duplicate (***p < 0.001; **p < 0.01).

Article Snippet: Cell culture and conditioned medium collection The OSCC cell lines (including SCC9, SCC15, SCC25, and CAL27; American Type Culture Collection (ATCC), USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Shanghai, China) containing 10% fetal bovine serum (FBS; Gibco, Shanghai, China), 100 U/mL penicillin (Gibco, Shanghai, China), and 100 μg/ mL streptomycin (Gibco, Shanghai, China).

Techniques:

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Rapamycin enhances the anti-angiogenesis and anti-proliferation ability of YM155 in oral squamous cell carcinoma.

doi: 10.1177/1010428317706213

Figure Lengend Snippet: Figure 2. The inhibitory effect of rapamycin and YM155 on the angiogenesis ability of OSCC cell lines. (a) The wound-healing assay was performed to examine the effects of rapamycin and YM155 on the OSCC-induced angiogenesis ability of the HUVECs. (b) Transwell assay was performed to examine the effects of rapamycin and YM155 on the OSCC-induced angiogenesis ability of the HUVECs. (c) Tube formation assay was performed to examine effects of rapamycin and YM155 on the OSCC-induced angiogenesis ability of the HUVECs. In these experiments, 100 nM of rapamycin and 60 nM of YM155 were used. All data are presented as mean ± SD from three different experiments performed in duplicate (**p < 0.01 vs the control group (no drug added); ††p < 0.01 the drug combination group vs the YM155 single group).

Article Snippet: Cell culture and conditioned medium collection The OSCC cell lines (including SCC9, SCC15, SCC25, and CAL27; American Type Culture Collection (ATCC), USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Shanghai, China) containing 10% fetal bovine serum (FBS; Gibco, Shanghai, China), 100 U/mL penicillin (Gibco, Shanghai, China), and 100 μg/ mL streptomycin (Gibco, Shanghai, China).

Techniques: Wound Healing Assay, Transwell Assay, Tube Formation Assay, Control

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Rapamycin enhances the anti-angiogenesis and anti-proliferation ability of YM155 in oral squamous cell carcinoma.

doi: 10.1177/1010428317706213

Figure Lengend Snippet: Figure 3. Effects of rapamycin and YM155 on the OSCC-induced angiogenesis ability. (a) The CAM assays as well as Matrigel plug assay were performed to examine the effects of rapamycin and YM155 on the OSCC-induced angiogenesis ability in vivo. The eggs and the nude mice were divided into five groups as before: Control group, conditioned medium without YM155 and rapamycin group, conditioned medium with YM155 single group, conditioned medium with rapamycin single group, conditioned medium with rapamycin and YM155 group. A volume of 100 nM of rapamycin and 60 nM of YM155 was used. (b) All data are presented as mean ± SD from three different experiments performed in duplicate and then analyzed by Graphpad (**p < 0.01 vs the control group (no drug added); ††p < 0.01 the drug combination group vs the YM155 single group).

Article Snippet: Cell culture and conditioned medium collection The OSCC cell lines (including SCC9, SCC15, SCC25, and CAL27; American Type Culture Collection (ATCC), USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Shanghai, China) containing 10% fetal bovine serum (FBS; Gibco, Shanghai, China), 100 U/mL penicillin (Gibco, Shanghai, China), and 100 μg/ mL streptomycin (Gibco, Shanghai, China).

Techniques: Matrigel Assay, In Vivo, Control

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Rapamycin enhances the anti-angiogenesis and anti-proliferation ability of YM155 in oral squamous cell carcinoma.

doi: 10.1177/1010428317706213

Figure Lengend Snippet: Figure 4. Rapamycin promotes the downregulation of VEGF and survivin of YM155 in OSCC cell lines through the inhibition of the phosphorylation of mTOR pathway. (a) The secretion level of VEGF in OSCC cell lines was examined by ELISAs after they were treated with 30 nM or 60 nM YM155 and 100 nM rapamycin for 12 h. (b) The expression levels of p-mTOR (Ser-2448) and p-S6 (Ser-235/236) and their corresponding total protein and the expression levels of survivin and VEGF were determined by western blot analysis. All data are presented as mean ± SD from three different experiments performed in duplicate (*p < 0.05 vs the control group (no drug added); †p < 0.05 the drug combination group vs the YM155 single group with the same dose of YM155).

Article Snippet: Cell culture and conditioned medium collection The OSCC cell lines (including SCC9, SCC15, SCC25, and CAL27; American Type Culture Collection (ATCC), USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Shanghai, China) containing 10% fetal bovine serum (FBS; Gibco, Shanghai, China), 100 U/mL penicillin (Gibco, Shanghai, China), and 100 μg/ mL streptomycin (Gibco, Shanghai, China).

Techniques: Inhibition, Phospho-proteomics, Expressing, Western Blot, Control