A novel mechanism of 6-methoxydihydroavicine in suppressing ovarian carcinoma by disrupting mitochondrial homeostasis and triggering ROS/ MAPK mediated apoptosis

Huachang Zhang; Fugen Shangguan; Lan Zhang; Nengfang Ma; Shuling Song; Li Ma; Chuntong Liu; Mengke Liu; Jing An; Hua Li; Qizhi Cao

doi:10.3389/fphar.2023.1093650

A novel mechanism of 6-methoxydihydroavicine in suppressing ovarian carcinoma by disrupting mitochondrial homeostasis and triggering ROS/ MAPK mediated apoptosis

Front Pharmacol. 2023 May 5:14:1093650. doi: 10.3389/fphar.2023.1093650. eCollection 2023.

Authors

Huachang Zhang¹, Fugen Shangguan², Lan Zhang³, Nengfang Ma⁴, Shuling Song⁵, Li Ma¹, Chuntong Liu¹, Mengke Liu¹, Jing An⁶, Hua Li³, Qizhi Cao¹

Affiliations

¹ Department of Immunology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong, China.
² Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
³ The Affiliated Taian City Central Hospital of Qingdao University, Taian, Shandong, China.
⁴ School of Life and Environmental Sciences, Wenzhou University, Wenzhou, China.
⁵ School of Gerontology, Binzhou Medical University, Yantai, Shandong, China.
⁶ Division of Infectious Diseases and Global Health, School of Medicine, University of California San Diego (UCSD), La Jolla, CA, United States.

Abstract

Introduction: Alkaloids derived from M. cordata (Papaveraceae family), have been found to display antineoplastic activity in several types of cancer. However, the antitumor effects and mechanisms of a new alkaloid extracted from the fruits of M. cordata, named 6-Methoxydihydroavicine (6-ME), remains unclear in the case of ovarian cancer (OC). Methods: CCK-8 assay was employed to analyze the cell viabilities of OC cells. RTCA, and colony-formation assays were performed to measure OC cell growth. Alterations in apoptosis and ROS levels were detected by flow cytometry in accordance with the instructions of corresponding assay kits. A Seahorse XFe96 was executed conducted to confirm the effects of 6-ME on cellular bioenergetics. Western blot and q-RT-PCR were conducted to detect alterations in target proteins. The subcutaneous xenografted tumor model of OC was used to further validate the anti-tumor activity of 6-ME in vivo. Results: Here, we reported for the first time that 6-ME inhibits OC cells growth in vitro and in vivo. Meanwhile, we found that 6-ME showed great antineoplastic activities by disrupting mitochondria homeostasis and promoting apoptosis in OC cells. Further investigation of the upstream signaling of apoptosis revealed that 6-ME-triggered apoptosis was induced by reactive oxygen species (ROS)-mediated mitogen-activated protein kinase (MAPK) activation and mitochondria dysfunction in OC cells. Furthermore, we found oxaloacetic acid (OAA), a crucial metabolite has been proved to be related to NADPH production, can block the cytotoxicity and accumulation of ROS caused by 6-ME in OC cells. Discussion: In summary, our data show that 6-ME exhibits cytotoxicity to OC cells in a ROS-dependent manner by interrupting mitochondrial respiration homeostasis and inducing MAPK-mediated apoptosis. This evidence suggests that 6-ME is a promising remedy for OC intervention.

Keywords: 6-methoxydihydroavicine (6-ME); MAPK; mitochondria homeostasis; ovarian cancer (OC); oxaloacetic acid (OAA) metabolism; reactive oxygen species (ROS).

Grants and funding

This work has been supported by the National Natural Science Foundation of China (Nos. 81672757 and 82200109), the Shandong Provincial Natural Science Foundation Joint Fund (ZR2021LZY024), the Youth Innovation and Technology Support Program in Colleges and Universities of Shandong Province (No. 2022KJ090), the Project of Zhejiang Provincial Naturel Science Foundation of China (LQ23H310006), and grants from the Science and Technology Bureau of Wenzhou (Y20220171).