Berbamine Hydrochloride inhibits lysosomal acidification by activating Nox2 to potentiate chemotherapy-induced apoptosis via the ROS-MAPK pathway in human lung carcinoma cells

• DOI: 10.1007/s10565-022-09756-8
• Publication Date: 2022-09-07T03:04:07Z
• Authors: Yujuan Zhan; Qiugu Chen; Yue Song; Xianli Wei; Tingxiu Zhao; Bonan Chen; Chengxi Li; Wenbo Zhang; Yanjun Jiang; Yuhui Tan; Biaoyan Du; Jianyong Xiao; Kun Wang
• Keywords: 0301 basic medicine; Lung Neoplasms; Carcinoma; Antineoplastic Agents; Apoptosis; Hydrogen-Ion Concentration; Cathepsins; 3. Good health; Mice; 03 medical and health sciences; Autophagy; Humans; Animals; Cisplatin; Reactive Oxygen Species; Lysosomes; Lung

Autophagy is typically activated in cancer cells as a rescue strategy in response to cellular stress (e.g., chemotherapy). Herein, we found that Berbamine Hydrochloride (Ber) can act as an effective inhibitor of the late stage of autophagic flux, thereby potentiating the killing effect of chemotherapy agents. Lung carcinoma cells exposed to Ber exhibited increased autophagosomes, marked by LC3-II upregulation. The increased level of p62 after Ber treatment indicated that the autophagic flux was blocked at the late stage. The lysosome staining assay and cathepsin maturation detection indicated impaired lysosomal acidification. We found that Nox2 exhibited intensified co-localization with lysosomes in Ber-treated cells. Nox2 is a key enzyme for superoxide anion production capable of transferring electrons into the lysosomal lumen, thereby neutralizing the inner protons; this might explain the aberrant acidification. This hypothesis is further supported by the observed reversal of lysosomal cathepsin maturation by Nox2 inhibitors. Finally, Ber combined with cisplatin exhibited a synergistic killing effect on lung carcinoma cells. Further data suggested that lung carcinoma cells co-treated with Ber and cisplatin accumulated excessive reactive oxygen species (ROS), which typically activated MAPK-mediated mitochondria-dependent apoptosis. The enhanced anti-cancer effect of Ber combined with cisplatin was also confirmed in an in vivo xenograft mouse model. These findings indicate that Ber might be a promising adjuvant for enhancing the cancer cell killing effect of chemotherapy via the inhibition of autophagy. In this process, Nox2 might be a significant mediator of Ber-induced aberrant lysosomal acidification.