Fig. 8 | Signal Transduction and Targeted Therapy

Fig. 8

From: Exosomal transfer of pro-pyroptotic miR-216a-5p exacerbates anthracycline cardiotoxicity through breast cancer-heart pathological crosstalk

Fig. 8

The miR-216a-5p/ITCH axis reduces TXNIP ubiquitination, aggravating DOX-induced cardiomyocyte pyroptosis. To determine whether miR-216a-5p inversely regulates TXNIP ubiquitination to promote TXNIP expression in DOX-treated adult murine ventricular cardiomyocytes (AMVCs), (a) TXNIP ubiquitination levels were assessed via Co-IP. AMVCs were transfected with a miR-216a-5p inhibitor or mimic for 24 hours and exposed to DOX or DMSO for another 24 hours. The protein lysates were immunoprecipitated with an anti-TXNIP antibody and immunoblotted with the indicated antibodies. To explore whether this effect could be regulated by ITCH knockdown or overexpression, (b) we assessed TXNIP ubiquitination via co-IP after AMVCs were transfected with adenovirus-shITCH or overexpression (oe) ITCH and transfected with the miR-216a-5p mimic. The reverse effect of TXNIP knockdown on DOX-induced pyroptosis was verified by (c) cell viability (n = 5), (d–f) LDH release; IL-18 and IL-1β levels; (g–k) and levels of pyroptosis-related proteins (NLRP3, GSDMD-N, cleaved-caspase-1, and cleaved-IL-1β) (n = 5). “ns” indicates non-significant, and “DOX” indicates doxorubicin. Data are presented as means ± SD

Back to article page