Efficacy of Quercetin-Sensitized Cisplatin

against N-Nitroso-N-Methylurea Induced

Testicular Carcinogenesis in Wistar Rats

Dr. sherif Refaat , MSc, MD

Lecturer of medical Oncology, OCMU
Faculty of Medicine
Mansoura University
Background
- Testicular cancer is one of the most common solid tumors in males and is considered a public health problem.
Both genetic and environmental factors contributed to the development of the testicular cancer
- N-nitosourea-N-methylurea (MNU) is used to cause cancer, such as breast cancer and other tumors in animal models.
- Quercetin is a natural bioflavenoid that has antioxidant , anti-inflammatory & anti-angiogenic effects.
- Quercetin-treatment in combination with cisplatin modulates gene expression of cyclins and cyclin dependent kinases . In
addition, affects the upregulation of genes involved in JNK, p38 and MAPK/ERK pathways coincides with decreased ERK
phosphorylations
- Quercetin causes downregulation of Wnt 16 expression in cisplatin – damaged tumor associated fibroblasts , which is a 1ry cause
for cisplatin resistance , hence increasing cisplatin sensitivity and effect.

Aim of the study:

The present study aimed to illustrate the therapeutic potential of quercetin and/ or cisplatin in the treatment MNU
induced testicular carcinogenesis.
Material and Methods:
In total 70 adult male albino rats were categorized into seven groups, control, quercetin-treatment (10 mg/kg body
weight), cisplatin-treatment (2 mg/kg. body weight), cisplatin and quercetin-treatment, MNU-treatment, MNU
plus quercetin-treatment and MNU plus quercetin and cisplatin-treatment.
- Treatment with quercetin and/or cisplatin was performed after 2 months of MNU induced testicular
carcinogenesis.
- The studied groups were euthanized and sacrificed and their testes were examined for :
1- Gene expression: extraction of testicular mRNA and testing of BAX, BCL2, MPO and SOD expression by RT-PCR.
2- Biochemical assay : of IL-6, AFP, caspase-3, testosterone , SOD & MDA.
3- Histological and IHC analysis for markers of inflammation and apoptosis of germ cells : Caspase-3 , TNF-alpha,
PCNA, P53.
Results:
1- Body weight gain and absolute and relative testis weight :
- Compared to the control, the gain in body weight in the MNU carcinogenesis group was significantly less than in the
cisplatin-treated group. Quercetin-treatment had less effect on MNU carcinogenesis than the cisplatin- treated group.
- The MNU carcinogenesis group demonstrated a decreased absolute and relative testis weight compared with the
cisplatin-toxicated group and varied markedly from the control values .
2- Morphometric assessment of testicular structure:
- Number of inactive ST : markedly increased in cisplatin intoxicated group compared to control group BUT, markedly
improved “decreased NO” by adding quercetin “but still < control.
- Mean diameter of ST : markedly decreased in MNU group . But increased by adding quercetin ±cisplatin “better in
quercetin alone “ , but still < control.
Results:
3- Histopathology:
 Results:
•4- Immunohistochemistry :
-proliferating cell nuclear antigen (PCNA):
markedly decreased staining in MNU and
cisplatin groups , and showed moderate
increased staining after adding quercetin.
- Caspase-3 , TNF-α, P53 : increased in
cisplatin &MNU ttt groups and decreased
staining by adding Quercetin.
5- biochemical investigations : in cisplatin and
MNU ttt groups:
*decreased testicular testosterone and SOD
*increased MDA , IL-6 & caspase-3 .
All changes were reversed by adding quercetin.
 Results:
6- Gene expression of BAX, BCL2, MPO and SOD by real time PCR:
In cisplatin & MNU ttt groups :
*increased gene expression of MPO and BAX
*decreased gene expression of BCL2 & SOD.
Which is reversed by adding quercetin ± cisplatin “ but still < control or quercetine alone”.
Discussion:
1- Our study showed decreased body weight gain and testicular weight “relative and absolute” in MNU and cisplatin intoxicated
groups.
2- cisplatin group showed:
A- increased testicular germ cell death confirmed by : * increased IHC for TNF-α, caspase-3 and decreased PNCA staining.
* increased gene expression of BAX & MPO and decreased BCL2
B- oxidative /antioxidant imbalance : with decreased SOD activity and gene expression , increased MDA level causing increased O2
free radical induced testicular damage and decreased testosterone levels.
3- Quercetin addition to cisplatin : partially reversed and improved all the previous deleterious effects of cisplatin
4- possible mechanism of quercetin :
- Anti-inflammatory effect : by decreasing release of cytokines , and inhibition of COX and LOX enzymes,
- Metabolized into quercetin 3- O glucuronide which has antioxidant effect and inhibition of capsase-3 and PARP leading to
decreased tumor cellular proliferation.
- Increased apoptosis in cancer cells “↓ AFP levels” due to increased expression of pro-apoptotic proteins.
- Anti-angiogenic effect.
Conclusion:

• quercetin showed therapeutic potential against testicular cancer and

its combination with the cisplatin-treatment resolved its cytotoxicity

and improved the antioxidant activity of the testicular cells and

warranting its germ cell cytoprotective action.

Thank
you