## 4 Discussion  
Multiple reports have demonstrated that peanut sprouts contain high protein content, unsaturated fatty acid content, polyphenic content, fiber content, and bioactive components [24; 25; 26]. Previous studies indicate that high levels of polyphenic compounds, such as caffe acid, coumaric acid, hesperidin, trans-arachidin-1, arabin 2, arabin 3, resveratrol, IPP, IPD, trans-arachidin-1, trans-arachidin-2 and trans-arachidin-3, are present in peanut sprouts [24; 25; 26]. Resverartrol and other phenolic compounds, such as coumaric acid, hesperidin and their respective stilbene derivatives, were induced during seed germination [24; 25]. We recently found that PSEFS generate high levels of resveratrol as compared with those cultivated with hydroponic technology [25; 26]. In the present study, we investigated the in vitro and in vivo antitumor effects of PSEFS in bladder cancer. The results demonstrated that PSEFS inhibited the proliferation of bladder cancer T24 cells without cell toxicity. Antitumor agents that mainly function by inhibiting cell cycle transition have previously been developed, as controlling cell cycle regulators responsible for cell cycle progression at specific stages represent promising candidates for the development of antitumor molecules [11; 12; 33; 34]. Previous studies demonstrated that several compounds exerted multiple actions in regulation of cell cycle in cancer cell lines [35; 36; 37]. In the present study, we investigated whether PSEFS affects cell cycle progression, with the results indicating that PSEFS treatment induced G1-phase arrest due to  
Figure 5: Antitumor efficacy of PSEFS in xenografted mice transplanted with T24 cells. The indicated concentrations of PSEFS (0. 20, and 200 mg/kg) and cisplatin (5 mg/kg; positive control) were orally administered to compare antitumor efficacy. (**A**) Appearance and quantification of weights of tumors separated from xenografted mice following treatment with PSEFS or cisplatin. (**B**) Tumor volumes were evaluated daily in xenografted mice. (**C**) Body weight loss was assessed during PSEFS administration and cisplatin treatment. (**D**) H&E and Ki-67 staining were performed to evaluate tumor growth. (**E**) After treatment with PSEFS, hematobiochemical markers (AST, ALT, and CK) were assessed in plasma from xenografted mice. Data represent the mean \(\pm\) SD (n = 5). * \(p<0.05\) vs. control. upregulation of the cell cycle inhibitor p21 WAF1 and downregulation of CDKs and cyclins in T24 cells. These findings demonstrated that PSEFS inhibited T24 cell proliferation by suppressing cyclin and CDK levels through induction of p21 WAF1 expression. This suggests that regulators associated with cell cycle progression might represent molecular targets for PSEFS-mediated clinical applications against bladder cancer. Previous studies demonstrated the important function of early signaling molecules in various cellular events attributed to cell proliferation and cell cycle progression [7; 8; 38]. Cumulative reports indicated that some compounds govern cell proliferation through regulation of MAPKs signaling pathway [39; 40]. However, information on the antitumor effect of PSEFS relevant to signaling pathways has not been reported. Therefore, we investigated the regulatory mechanism associated with PSEFS-mediated inhibition of T24 cell proliferation and the related signaling pathways involving including AKT and MAPKs (p38 MAPK, JNK, and ERK1/2). The results revealed that PSEFS treatment induced phosphorylation of p38 MAPK. SB203580 treatment attenuated PSEFS-induced inhibition of T24 cell proliferation, suggesting p38 MAPK as a factor partially mediated by suppression of T24 cell proliferation. Additionally, we demonstrated that PSEFS treatment suppressed T24 cell invasion and migration associated with downregulation of MMP-9 activity in the presence of PSEFS.