In vivo and in vitro effects of RAD001 on bladder cancer

Abstract

Objective: To evaluate the influence of Everolimus (RAD001) on chemically induced urothelial lesions in mice and its influence on in vitro human bladder cancer cell lines.Methods: ICR male mice were given N-butyl-N-(4-hydroxybutyl) nitrosamine in drinking water for a period of 12 weeks. Subsequently, RAD001 was administered via oral gavage, for 6 weeks. At the end of the experiment, all the animals were sacrificed and tumor development was determined by means of histopathologic evaluation; mammalian target of rapamycin (mTOR) expressivity was evaluated by immunohistochemistry. Three human bladder cancer cell lines (T24, HT1376, and 5637) were treated using a range of RAD001 concentrations. MTT assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and flow cytometry were used to assess cell proliferation, apoptosis index, and cell cycle analysis, respectively. Immunoblotting analysis of 3 cell line extracts using mTOR and Akt antibodies was performed in order to study the expression of Akt and mTOR proteins and their phosphorylated forms.Results: The incidence of urothelial lesions in animals treated with RAD001 was similar to those animals not treated. RAD001 did not block T24 and HT1376 cell proliferation or induce apoptosis. A reduction in cell proliferation rate and therefore G 0 /G 1 phase arrest, as well as a statistically significant induction of apoptosis (P ϭ 0.001), was only observed in the 5637 cell line.Conclusion: RAD001 seems not to have a significant effect on chemically induced murine bladder tumors. The effect of RAD001 on tumor proliferation and apoptosis was achieved only in superficial derived bladder cancer cell line, no effect was observed in invasive cell lines.

Altered expression of CKs 14/20 is an early event in a rat model of multistep bladder carcinogenesis

“…The rat and mouse models are complementary: while rats typically develop papillary tumours in response to BBN, mice develop invasive lesions (Vasconcelos‐Nóbrega et al . ; Oliveira et al . ).…”

“…Other models, relying on bladder cancer cell lines xenografted into immunodeficient mice, lack these characteristics. The rat and mouse models are complementary: while rats typically develop papillary tumours in response to BBN, mice develop invasive lesions (Vasconcelos-N obrega et al 2013;Oliveira et al 2014). Now, for the first time, we traced the expression of CK14 and CK20 during the consecutive steps of BBN-induced carcinogenesis in the rat bladder.…”

Ruthenium porphyrin-induced photodamage in bladder cancer cells

“…Preclinical studies also play an important role in this aspect [32]. Investigations on bladdercancer cell lines is a natural first step to determine the properties of newly developed drugs and to test various treatment strategies [6,[33][34][35][36].…”

Synergistic Effect between Cisplatin and Sunitinib Malate on Human Urinary Bladder-Cancer Cell Lines

“…Thus, novel targeted therapies are sorely required to further improve the effectiveness of urinary bladder cancer chemotherapy. Preclinical models play a crucial role in this setting [17] and urinary bladder-cancer cell lines have been invaluable research tools to evaluate the efficacy of new drugs [18–20]. In the present study, we investigated if sunitinib malate could strengthen cisplatin cytotoxicity, using as in vitro models three human urinary bladder-cancer cell lines representative of human urinary bladder tumors: one nonmuscle invasive cell line (5637) and two muscle-invasive cell lines (T24 and HT1376).…”