## 4 Discussion
Several reports indicate MMP-9 activity as essential for the migratory and invasive potential of bladder cancer cells, as MMP-9 induces degradation of the extracellular matrix and promotes progression of tumor cell invasion and migration [6; 14; 15]. In the present study, we showed that PSEFS repressed T24 cell invasion and migration by reducing MMP-9 activity and expression, specifically through control of AP-1, NF-\(\kappa\)B, and SP1 targeting of its promoter region [16; 17; 18]. Further investigation of the DNA-binding activity of these transcription factors confirmed that PSEFS treatment decreased their DNA-binding abilities to the MMP-9 promoter region in T24 cells, revealing that PSEFS-mediated suppression of T24 cell invasion and migration is associated with reduced MMP-9 expression and activity as a consequence of decreased transcriptional activation by SP1, NF-\(\kappa\)B, and AP-1. We then evaluated the antitumor efficacy of PSEFS in vivo using a mouse xenograft model. PSEFS administration reduced both tumor volume and weight in xenografted mice bearing bladder T24 tumors, with decreased tumor cell proliferation confirmed by H&E and Ki-7 staining, and establishment of an effective PSEFS dose at 200 mg/kg. Importantly, we identified the antitumor efficacy of 20 mg/kg PSEFS as equivalent value to that of 5 mg/kg cisplatin, although without the observed side effects (i.e., significant loss of body weight or hematobiochemical activity (AST, ALT, and CK)) observed following cisplatin treatment, thereby underscoring the lack of PSEFS-related toxicity. These findings indicated that resveratrol-enriched PSEFS can be utilized for the treatment and prevention of bladder cancer with minimal side effects or natural supplements, although additional studies incorporating detailed toxicity tests for antitumor agents need to be performed. Our study demonstrates the antitumor efficacy of PSEFS against bladder cancer. PSEFS-induced inhibition of bladder cancer T24 cell proliferation mainly involved in the promotion of G1-phase cell cycle arrest through downregulation of CDKs and cyclins was caused by an increased expression of p21 WAF1. Additionally, p38 MAPK phosphorylation increased with PSEFS treatment. The PSEFS-mediated suppressive effect of migration and invasion potential in T24 cells was due to a decreased binding of transcription factors associated with MMP-9 regulation. Moreover, administration of PSEFS abolished tumor growth rate in xenograft mice bearing T24 tumor cells without noticeable side effects. These data suggest PSEFS as an effective and promising candidate antitumor molecule against bladder cancer. The following are available online at [[http://www.mdpi.com/2076-3417/10/23/8758/s1](http://www.mdpi.com/2076-3417/10/23/8758/s1)]([http://www.mdpi.com/2076-3417/10/23/8758/s1](http://www.mdpi.com/2076-3417/10/23/8758/s1)). [PERSON] and [PERSON] planned and designed the experiments. [PERSON], [PERSON], and [PERSON] performed the experiments. [PERSON] and [PERSON] analyzed the data. [PERSON] and [PERSON] wrote the manuscript. [PERSON], [PERSON], [PERSON], and [PERSON] reviewed and discussed the results. All authors have read and agreed to the published version of the manuscript. **Funding:** This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (NRF-2018R1A6A1A03025159). This research was also supported by the Chung-Ang University Research Scholarship Grant in 2019. **Conflicts of Interest:** The authors declare no conflict of interest.