Tanshinone I inhibits metastasis of cervical cancer cells by inducing BNIP3/NIX-mediated mitophagy and reprogramming mitochondrial metabolism

Shuna Cui; Tingting Chen; Mengmeng Wang; Yuanyuan Chen; Qi Zheng; Xinyi Feng; Shihua Li; Junsong Wang

doi:10.1016/j.phymed.2022.153958

Tanshinone I inhibits metastasis of cervical cancer cells by inducing BNIP3/NIX-mediated mitophagy and reprogramming mitochondrial metabolism

Phytomedicine. 2022 Apr:98:153958. doi: 10.1016/j.phymed.2022.153958. Epub 2022 Jan 29.

Authors

Shuna Cui¹, Tingting Chen², Mengmeng Wang², Yuanyuan Chen², Qi Zheng³, Xinyi Feng², Shihua Li⁴, Junsong Wang⁵

Affiliations

¹ Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Medical College of Yangzhou University, Jiangyang Middle Road 136, Yangzhou 225001, China; Department of Gynecology And Obstetrics, Affiliated Hospital of Yangzhou University, Yangzhou, 225000, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou, 225000, China. Electronic address: sncui@yzu.edu.cn.
² Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Medical College of Yangzhou University, Jiangyang Middle Road 136, Yangzhou 225001, China.
³ School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Xiao Ling Wei No. 200, Nanjing, 210094, China.
⁴ Department of Gynecology And Obstetrics, Affiliated Hospital of Yangzhou University, Yangzhou, 225000, China.
⁵ School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Xiao Ling Wei No. 200, Nanjing, 210094, China. Electronic address: wang.junsong@gmail.com.

PMID: 35124382
DOI: 10.1016/j.phymed.2022.153958

Abstract

Background: Cervical cancer is the most common malignancy of the female lower genital tract. Tanshinone I (Tan I) is one of the crucial lipid-soluble components of red sage (Salvia miltiorrhiza). While its mode of action against cervical cancer is unclear.

Purpose: Our study aimed to explore the role of Tan I on cervical cancer in vitro.

Study design and methods: Effects of Tan I on cervical cancer cells viability, migration and mitochondrial function were investigated by Cell Counting Kit-8, Transwell and Fluorescence laser confocal microscope assays respectively. The potential mechanism of Tan I was uncovered by an integrative approach combining RNA profiling and hydrogen nuclear magnetic resonance-based metabolic analysis, molecular docking and Western blot.

Results: Tan I significantly inhibited the growth and colony formation of HeLa and SiHa cells. It induced apoptosis and cell cycle S phase arrest at low (12.5-25 μM) but not high (50 μM) concentrations. It also altered the HeLa cell ultrastructure, decreased the membrane potential and increased the total mitochondrial content. Further, Tan I induced autophagic flux and the colocalization of mitochondria with lysosomes, led to decreased adhesion, invasion, and migration of cervical cancer cells. Transcriptomic analysis revealed that Tan I altered the RNA profile and signal processing in HeLa cells. Tan I significantly impacted "central carbon metabolism in cancer" and "mitophagy-animal" processes. A global metabolic analysis identified 25 metabolites affected by Tan I treatment in HeLa cells. Changes in the metabolic profile indicated that Tan I affected such processes as protein digestion and absorption, central carbon metabolism in cancer, and aminoacyl-tRNA biosynthesis in cervical cancer cells. Furthermore, Tan I significantly induced the expression of mitophagy-related proteins BNIP3, NIX and Optineurin and the conversion from LC3-I to LC3-II, inhibited the NDP52 and P62 level in a concentration-dependent manner. While CQ further increased the conversion of LC3-I to LC3-II and the expression of P62. Moreover, Tan I interacted with BNIP3 and NIX through hydrogen bond. Tan I induce mitophagy could be prevented by BNIP3 and NIX siRNA transfection.

Conclusion: Tan I induced the BNIP3/NIX-mediated mitophagy, and reprogrammed the mitochondrial metabolism in cervical cancer cells, thus inhibiting metastasis.

Keywords: Cervical cancer; Mitochondrial metabolism; Mitophagy; Tanshinone I.