Effects of cold atmospheric-pressure plasma in combination with doxorubicin drug against breast cancer cells in vitro and in vivo

Cold atmospheric plasma (CAP) has been suggested for medical applications that can be applied indirectly through plasma-activated medium (PAM) and recently it has been introduced as an innovative therapeutic approach for all cancer types. Studies have exhibited that ROS/RNS are key factors in CAP-dependent apoptosis; nevertheless, ROS/RNS stability are weak. Combination therapy is considered an effective strategy to overcome these problems. In the present research, we revealed that the combination of CAP and doxorubicin (DOX) significantly induces the apoptosis of breast cancer cells both in vitro and in vivo. Our results indicated that both Ar and He/O2 CAP treatment as well as DOX drug alone reduced cell growth. CAP/PAM treatment in combination with DOX induced apoptosis in MCF-7 breast cancer cells and 4T1-implanted BALB/c mice, resulting in a significant increase in antitumor activity. The apoptotic effects of CAP-DOX on MCF-7 cells were inferred from altered expression of BAX and cleaved-caspase-3 which mechanistically take place through the mitochondrial pathway mediated by Bcl-2 family members. Besides, the BAX/BCL-2 ratio is significantly higher in the simultaneous treatment of CAP and DOX. This ratio was equal to 2.82 +/- 0.24, 2.54 +/- 0.30, and 11.27 +/- 0.31 for treatment with DOX, He/O2 plasma, and combination treatment, respectively. Additionally, the tumor growth rate of He/O2-PAM + DOX and Ar-PAM + DOX treatments was significantly inhibited by PAM-injection, and the tumor growth rate of PAM alone or DOX alone was slightly reduced. It can be concluded that the effect of PAM + DOX may increase the anticancer activity and decrease the dose required for the chemotherapeutic treatment.