Downregulation of topoisomerase 1 and 2 with acriflavine sensitizes bladder cancer cells to cisplatin-based chemotherapy

• Published in: Molecular biology reports 2022-04, Vol.49 (4), p.2755-2763
• Main Authors: Zargar, Parisa, Koochakkhani, Shabnaz, Hassanzadeh, Marziyeh, Ashouri Taziani, Yaghoub, Nasrollahi, Hamid, Eftekhar, Ebrahim
• Format: Article
• Language: English
• Subjects: Acriflavine; Acriflavine - pharmacology; Acriflavine - therapeutic use; Animal Anatomy; Animal Biochemistry; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Antitumor activity; Apoptosis; Bcl-2 protein; Biomedical and Life Sciences; Bladder cancer; Cell culture; Cell Line, Tumor; Chemotherapy; Cisplatin; Cisplatin - pharmacology; Cisplatin - therapeutic use; Down-Regulation; Drug dosages; Drug Resistance, Neoplasm; Gene expression; Histology; Humans; Hypoxia-inducible factor 1a; Life Sciences; Morphology; Original Article; p53 Protein; Urinary Bladder Neoplasms - drug therapy; Urinary Bladder Neoplasms - genetics; Urinary Bladder Neoplasms - pathology
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-021-07087-1

Background Resistance to cisplatin is a major obstacle to effective treatment of bladder cancer (BC). The present study aimed to determine whether a combination of acriflavine (ACF) with cisplatin could potentiate the antitumor property of cisplatin against the BC cells. Furthermore, the molecular mechanism behind the anticancer action of ACF was considered. Methods and results Two human BC cells (5637 and EJ138) contain mutated form of p53 was culture in standard condition. Cotreatment protocol (simultaneous combination of IC 30 value of ACF + various dose of cisplatin for 72 h) and pretreatment protocol (pretreatment with IC 15 value of ACF for 24 h + various dose of cisplatin for 48 h) was used to determine the effect of ACF on the cells’ sensitivity to main drug cisplatin. To assess the mechanism of action of ACF, real-time PCR was used to evaluate mRNA levels of hypoxia-inducible factor-1α (HIF-1α), Bax, Bcl-2, topoisomerase1 (TOP1) and topoisomerase 2 (TOP2A). Combination of ACF with cisplatin either as cotreatment or opretreatment protocol could significantly reduce the IC 50 values of cisplatin as compared to the IC 50 of cisplatin when use as a single drug. In addition, ACF could markedly decrease mRNA expression of TOP1 and TOP2A without changing the expression of HIF-1ɑ, Bax and Bcl-2. Conclusions Our findings indicate that combination of cisplatin with ACF was able to significantly enhance the sensitivity of BC cells to cisplatin. The antitumor activity of ACF is exerted through the downregulation of TOP1 and TOP2A genes expression. ACF may serve as an adjuvant to boost cisplatin-based chemotherapy.