BDNF       Higher expression (n = 41) Lower expression (n = 24) p-value Distribution Solitary 10 15 *0.002   Multiple 31 9   Differentiation Well 23 7 *0.036   Moderate-poor 18 17   Stage I+II 7 12 *0.005   III 34 12   Lymph node metastasis + – 19 22 4 20 *0.016 * = statistically significant difference. Table 2 Clinicopathological

characteristics and TrkB expression by immunohistochemistry in 65 cases of HCCs.     TrkB       Positive expression www.selleckchem.com/products/AZD6244.html (n = 36) Negative expression (n = 29) p-value Distribution Solitary 10 15 *0.049   Multiple 26 14   Differentiation Well 20 10 0.090   Moderate-poor 16 19   Stage I+II 6 13 *0.013   III 30 16   Lymph node metastasis + – 14 22 9 20 0.510 * = statistically significant difference. The secretion of BDNF in HepG2 and this website HCCLM3 cells by ELISA BDNF is a cytokine secreted by a few

human cancers, supporting growth and survival of tumor cells [23]. To explore whether HCC cells express BDNF secretorily, BDNF in the supernatant of HepG2 and HCCLM3 cells was examined by ELISA assays. The amounts of BDNF produced extracellularly by HepG2 and HCCLM3 cells were 88.6 ± 14.4 pg/ml and 138.4 ± 22.2 pg/ml, respectively (p = 0.031), which was shown in Table 3. This result showed that HCCLM3 cells had more BDNF production, which probably correlated with its high metastatic potential. Table 3 Secretion of BDNF in supernatant of HepG2 and HCCLM3 cells PND-1186 clinical trial by ELISA. Cells BDNF concentration (pg/ml) p value HepG2 88.6 ± 14.4 *0.031 HCCLM3 138.4 ± 22.2   * = statistically significant difference. Anti-BDNF or K252a promoted cell apoptosis It was demonstrated BDNF/TrkB protected various tumor cells from apoptosis [24]. To investigate a positive role of BDNF/TrkB in HCC cell survival, apoptosis was examined

after anti-BDNF or K252a treatment using Annexin V-FITC assay by flow cytometry. The apoptotic rates of control, anti-BDNF and K252a treated HepG2 at 24 h time next point were 5.29 ± 0.54%, 20.21 ± 1.54%, 18.39 ± 0.83%, respectively (p = 0.000, Figure 2). And the apoptotic rates of control, anti-BDNF and K252a treated HCCLM3 at 24 h time point were 10.88 ± 0.42%, 30.35 ± 1.60%, 31.37 ± 2.16%, respectively (p = 0.000, Figure 2). These results suggested that neutralizing antibody specific for BDNF or Trk tyrosine kinase inhibitor K252a against TrkB probably antagonized the protection of BDNF/TrkB for HCC cells. Figure 2 Anti-BDNF or K252a treatment promoted cell apoptosis. The apoptotic cells in anti-BDNF or K252a group were apparently increased in HepG2 or HCCLM3, in contrast to those control cells. The results were indicated as mean ± SD of three individual tests. Effect of anti-BDNF or K252a on cell invasion To understand the potential signaling induced by BDNF/TrkB that affects cell invasion, anti-BDNF or K252a was used and the invasion of treated cells was examined by Transwell assay.