Efficient delivery of active proteins to specific cells and organs is one of the most important issues in medical applications. However, in most cases, proteins without appropriate carriers face numerous barriers when delivered to the target, due to their unsatisfied properties, such as poor stability, short half-life, and low membrane permeability. Herein, we have presented a large-pore mesoporous silica nanoparticle (LPMSN)-based protein delivery system. LPMSNs were obtained with ethyl acetate as a pore expander. A 2,3-dimethylmaleamic acid-containing silane coupling agent was modified on LPMSNs to provide pHtriggered charge reversal. After Cytochrome c (CC) was encapsulated in the large pores of LPMSNs, amino-terminated polyethylene glycol-modified gold nanoparticles (AuNPs) served as gateguards to cap the tunnels of LPMSNs and to avoid the leakage of CC. Above nanocomposites exhibited the capability to deliver active CC into cancer cells, charge reversal-induced protein release, as well as to initiate the apoptosis machinery of cancer cells in vitro. Importantly, the nanocomposites significantly inhibited tumor growth and extended survival rate without obvious side effects. This study provides a smart and efficient protein delivery platform with good safety profiles for efficacious tumor protein therapy in vivo. (c) 2021 Elsevier Inc. All rights reserved.