Pharmacological Inhibition of Endogenous Hydrogen Sulfide Attenuates Breast Cancer Progression

Nazeer Hussain Khan; Di Wang; Wenkang Wang; Muhammad Shahid; Saadullah Khattak; Ebenezeri Erasto Ngowi; Muhammad Sarfraz; Xin-Ying Ji; Chun-Yang Zhang; Dong-Dong Wu

doi:10.3390/molecules27134049

Pharmacological Inhibition of Endogenous Hydrogen Sulfide Attenuates Breast Cancer Progression

Molecules. 2022 Jun 23;27(13):4049. doi: 10.3390/molecules27134049.

Authors

Nazeer Hussain Khan^{1

2}, Di Wang¹, Wenkang Wang³, Muhammad Shahid⁴, Saadullah Khattak^{1

2}, Ebenezeri Erasto Ngowi^{1

5}, Muhammad Sarfraz^{1

6}, Xin-Ying Ji^{1

7}, Chun-Yang Zhang^{8

9}, Dong-Dong Wu^{1

10}

Affiliations

¹ Henan International Joint Laboratory for Nuclear Protein Regulation, School of Basic Medical Sciences, Henan University, Kaifeng 475004, China.
² School of Life Sciences, Henan University, Kaifeng 475004, China.
³ Department of Breast Surgery, The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.
⁴ Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Malaysia.
⁵ Department of Biological Sciences, Faculty of Science, Dar es Salaam University College of Education, Dar es Salaam 11101, Tanzania.
⁶ Faculty of Pharmacy, The University of Lahore, Lahore 54590, Pakistan.
⁷ Kaifeng Key Laboratory of Infection and Biological Safety, College of Medicine, Henan University, Kaifeng 475004, China.
⁸ Department of Thoracic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
⁹ Department of General Thoracic Surgery, Hami Central Hospital, Hami 839000, China.
¹⁰ School of Stomatology, Henan University, Kaifeng 475004, China.

Abstract

Hydrogen sulfide (H₂S), a gaseous signaling molecule, is associated with the development of various malignancies via modulating various cellular signaling cascades. Published research has established the fact that inhibition of endogenous H₂S production or exposure of H₂S donors is an effective approach against cancer progression. However, the effect of pharmacological inhibition of endogenous H₂S-producing enzymes (cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (3-MPST)) on the growth of breast cancer (BC) remains unknown. In the present study, DL-propargylglycine (PAG, inhibitor of CSE), aminooxyacetic acid (AOAA, inhibitor of CBS), and L-aspartic acid (L-Asp, inhibitor of 3-MPST) were used to determine the role of endogenous H₂S in the growth of BC by in vitro and in vivo experiments. An in silico study was also performed to confirm the results. Corresponding to each enzyme in separate groups, we treated BC cells (MCF-7 and MDA-MB-231) with 10 mM of PAG, AOAA, and L-Asp for 24 h. Findings reveal that the combined dose (PAG + AOAA + L-Asp) group showed exclusive inhibitory effects on BC cells' viability, proliferation, migration, and invasion compared to the control group. Further, treated cells exhibited increased apoptosis and a reduced level of phospho (p)-extracellular signal-regulated protein kinases such as p-AKT, p-PI3K, and p-mTOR. Moreover, the combined group exhibited potent inhibitory effects on the growth of BC xenograft tumors in nude mice, without obvious toxicity. The molecular docking results were consistent with the wet lab experiments and enhanced the reliability of the drugs. In conclusion, our results demonstrate that the inhibition of endogenous H₂S production can significantly inhibit the growth of human breast cancer cells via the AKT/PI3K/mTOR pathway and suggest that endogenous H₂S may act as a promising therapeutic target in human BC cells. Our study also empowers the rationale to design novel H₂S-based anti-tumor drugs to cure BC.

Keywords: apoptosis; breast cancer; endogenous hydrogen sulfide; signaling pathway; tumor growth.

MeSH terms

Animals
Breast Neoplasms* / drug therapy
Cystathionine
Cystathionine beta-Synthase / metabolism
Cystathionine gamma-Lyase / metabolism
Female
Humans
Hydrogen Sulfide* / metabolism
Hydrogen Sulfide* / pharmacology
Mice
Mice, Nude
Molecular Docking Simulation
Phosphatidylinositol 3-Kinases
Proto-Oncogene Proteins c-akt
Reproducibility of Results
TOR Serine-Threonine Kinases

Substances

Cystathionine
Proto-Oncogene Proteins c-akt
TOR Serine-Threonine Kinases
Cystathionine beta-Synthase
Cystathionine gamma-Lyase
Hydrogen Sulfide

Grants and funding

This work was supported by grants from the National Natural Science Foundation of China (Nos. 81802718, U1504817); the Training Program for Young Backbone Teachers of Institutions of Higher Learning in Henan Province, China (No. 2020GGJS038); the Natural Science Foundation of Edu-cation Department of Henan Province, China (No. 21A310003); and the Foundation of Science and Technology Department of Henan Province, China (Nos. 222102310490, 222102310495).