Artemisinin pre-treatment fore cisplatin dosage enhances high grade urothelial carcinoma treatment in male albino mice via reverse gene expression modulation of FGFR3, HRAS, P53 and KDM6A

Abstract

Background: Urinary bladder cancer, is the 10th most common global cancer, diagnosed in over 600,000 people causing 200,000 deaths annually. Artemisinin and its derivatives are safe compounds that have recently been proven to possess potent anti-tumor effects in vivo, through inhibition of cancer cell growth. The aim of this study is to assess the efficiency of artemisinin as a cancer treatment alone and as a pre-treatment fore cisplatin therapy for high grade urothelial carcinoma.

Methods: Sixty male albino mice were divided into six groups, and BBN was used to induce urinary bladder cancer. Blood samples were tested for renal functions and complete blood counts, kidney and urinary bladder tissues were harvested for histopathological examination. Total RNAs from urinary bladder tissues was collected, and gene expression of FGFR3, HRAS, P53, and KDM6A was quantified using qRT-PCR.

Results: Compared to the induced cancer group, the results revealed that FGFR3 expression levels were down-regulated in the induced cancer group treated by artemisinin only and the induced cancer group pre-treated with artemisinin prior to cisplatin by ~ 0.86-fold and 0.4-folds, respectively, aligning with HRAS down-regulation by ~ 9.54-fold and 9.05-fold, respectively. Whereas, P53 expression levels were up-regulated by ~ 0.68-fold and 0.84-fold, respectively, in parallel with KDM6A expression, which is up-regulated by ~ 0.95-folds and 5.27-folds, respectively. Also, serum creatinine and urea levels decreased significantly in the induced cancer group treated by artemisinin alone and the induced cancer group pre-treated with artemisinin prior to cisplatin, whereas the induced cancer group treated by cisplatin their levels increased significantly. Moreover, Hb, PLT, RBC, and WBC counts improved in both cancer groups treated by artemisinin alone and pre-treated with artemisinin prior to cisplatin. Histologically, in kidney tissues, artemisinin pre-treatment significantly reduced renal injury caused by cisplatin. While Artemisinin treatment for cancer in bladder tissues reverted invasive urothelial carcinoma to moderate urothelial dysplasia.

Conclusions: This study indicates that artemisinin demonstrated a significant effect in reversal of the multi-step carcinogenesis process of high grade urothelial carcinoma and could enhance the effect of cisplatin therapy using artemisinin pre-treatment.

Keywords: Artemisinin; BBN; Cisplatin; FGFR3; HRAS; High grade urothelial carcinoma; KDM6A; P53; Urinary bladder cancer.

Fig. 1

Biochemical and hematological analysis in studied groups. A urea levels, (B) creatinine levels, (C) Hb concentrations, (D) RBC count, (E) PLT count and (F) WBC count in. Data are expressed as mean ± SD where, #P < 0.05 vs Control, *P < 0.05 vs BBN

Fig. 2

Histopathology of BBN-induced bladder cancer in mice. A A graphical illustration of the BBN treatment chronology. B Representative images demonstrating histological alterations. Where, (a) unremarkable urothelial lining with no changes, (b) hyperplastic urothelial lining with no atypia, (c) mild urothelial dysplasia, (d) moderate dysplasia with broad papillae lined by thickened urothelial lining, (e) full thickness dysplasia in the urothelial lining cells amounting to carcinoma in situ and (f) focus of invasive carcinoma nest in the lamina propria (red arrow)

Fig. 3

Histopathological examinations of kidney (a) and urinary bladder (b) tissues in studied groups. Where, (A) control group, (B) BBN (cancer group), (C) artemisinin group, (D) BBN + artemisinin group, (E) BBN + cisplatin group and (F) BBN + artemisinin + cisplatin group

Fig. 4

RT-qPCR was applied to detect the expressions of oncogenes (A) FGFR3, (B) HRAS and tumor suppressor genes (C) P53, (D) KDM6A in mouse urinary bladder tissues. Where, #P < 0.05 vs Control, *P < 0.05 vs BBN