Abstract

Peanut sprout extracts reportedly exhibit numerous beneficial effects; however, there are few investigations on the biological effects of peanut sprout extracts cultivated with fermented sawdust medium (PSEFS). Here, we examined whether PSEFS demonstrates antitumor activity against bladder cancer, in vitro and in vivo. The results showed that PSEFS prohibited the proliferation of bladder cancer T24 cells, with this effect attributed to induction of cell cycle arrest at the G1 phase through reduced expression of cyclins and cyclin-dependent kinases caused by a promotion of p21WAF1 expression. Additionally, PSEFS induced phosphorylation of p38 mitogen-activated protein kinase. Moreover, PSEFS treatment attenuated the invasive and migratory potential of T24 cells due to decreased matrix metalloproteinase-9 activity combined with downregulation of the transcriptional binding activity of SP1, activator protein -1, and nuclear factor-kappaB. Furthermore, PSEFS (20 mg/kg) attenuated the tumor-growth rate in xenograft mice bearing T24 cells, with an effect equivalent to that of cisplatin and in the absence of toxicity following weight-loss evaluation and hematobiochemical testing of PSEFS-treated mice. These results demonstrated the antitumor efficacy of PSEFS both in vitro and in vivo, thereby reporting it as a potential candidate for development of novel agents against bladder cancer.

Highlights

  • Bladder tumors are among the most fatal urological cancers worldwide, with 2 million recorded deaths in 2018 [1]

  • Generation of fluorescence-activated cell sorting (FACS) histograms to determine whether PSEFS-induced anti-proliferative effects in T24 cells were related to changes in cell cycle progression indicated that PSEFS treatment induced G1-phase arrest (Figure 1D–H)

  • The results showed that PSEFS treatment reduced the expression treated T24 cells, we evaluated the levels of cell cycle-related proteins positively modulating the level of CDK4 and its counterpart cyclin D1 in T24 cells (Figure 2A)

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Summary

Introduction

Bladder tumors are among the most fatal urological cancers worldwide, with 2 million recorded deaths in 2018 [1]. Dysregulation of tumor cell proliferation results from a progression of the cell cycle at the G1, S, and G2/M phases, with transition from G1 to S phase controlled by cell cycle regulators [9,10] These regulators include cyclins, cyclin-dependent kinases (CDKs), and p21WAF1 , which have recently received attention as primary targets for antitumor molecules [11,12]. We examined for the first time the antitumor efficacy of PSEFS against bladder cancer both in vitro and in vivo. These results may provide critical information for developing preventive or therapeutic resources for bladder cancer

Materials
Sample Preparation
Cell Culture
Cell Cycle Analysis
Immunoblots and IP
Apoptosis Assay
Wound-Healing Migration Assay
Boyden Chamber Invasion Assay
2.10. Zymography
2.11. Nuclear Extracts and EMSA
2.12. Mouse Xenograft Inoculation
2.13. Immunohistochemistry
2.14. Plasma Preparation and Biochemical Analysis
2.15. Statistical Analysis
Results
PSEFS promotes the phosphorylation
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