TY - JOUR
T1 - A High Dose of Calcitriol Inhibits Glycolysis and M2 Macrophage Polarization in the Tumor Microenvironment by Repressing mTOR Activation
T2 - in vitro and Molecular Docking Studies
AU - Munir, Maliha Tabassum
AU - Halim, Sobia Ahsan
AU - Santos, Julianna Maria
AU - Khan, Faizullah
AU - Khan, Ajmal
AU - Rahman, Md Mizanur
AU - Hussain, Fazle
AU - Al-Harrasi, Ahmed
AU - Gollahon, Lauren S.
AU - Rahman, Shaikh Mizanoor
N1 - © Copyright by the Author(s). Published by Cell Physiol Biochem Press.
PY - 2023/4/12
Y1 - 2023/4/12
N2 - BACKGROUND/AIMS: Macrophages interact with tumor cells within the tumor microenvironment (TME), which plays a crucial role in tumor progression. Cancer cells also can instruct macrophages to facilitate the spread of cancer and the growth of tumors. Thus, modulating macrophages-cancer cells interaction in the TME may be therapeutically beneficial. Although calcitriol (an active form of vitamin D) has anticancer properties, its role in TME is unclear. This study examined the role of calcitriol in the regulation of macrophages and cancer cells in the TME and its influence on the proliferation of breast cancer cells. METHODS: We modeled the TME, in vitro, by collecting conditioned medium from cancer cells (CCM) and macrophages (MCM) and culturing each cell type separately with and without (control) a high-dose (0.5 µM) calcitriol (an active form of vitamin D). An MTT assay was used to examine cell viability. Apoptosis was detected using FITC (fluorescein isothiocyanate) annexin V apoptosis detection kit. Western blotting was used to separate and identify proteins. Quantitative real-time PCR was used to analyze gene expression. Molecular docking studies were performed to evaluate the binding type and interactions of calcitriol to the GLUT1 and mTORC1 ligand-binding sites. RESULTS: Calcitriol treatment suppressed the expression of genes and proteins implicated in glycolysis (GLUT1, HKII, LDHA), promoted cancer cell apoptosis, and reduced viability and Cyclin D1gene expression in MCM-induced breast cancer cells. Additionally, calcitriol treatment suppressed mTOR activation in MCM-induced breast cancer cells. Molecular docking studies further showed efficient binding of calcitriol with GLUT1 and mTORC1. Calcitriol also inhibited CCM-mediated induction of CD206 and increased TNFα gene expression in THP1-derived macrophages. CONCLUSION: The results suggest that calcitriol may impact breast cancer progression by inhibiting glycolysis and M2 macrophage polarization via regulating mTOR activation in the TME and warrants further investigation in vivo.
AB - BACKGROUND/AIMS: Macrophages interact with tumor cells within the tumor microenvironment (TME), which plays a crucial role in tumor progression. Cancer cells also can instruct macrophages to facilitate the spread of cancer and the growth of tumors. Thus, modulating macrophages-cancer cells interaction in the TME may be therapeutically beneficial. Although calcitriol (an active form of vitamin D) has anticancer properties, its role in TME is unclear. This study examined the role of calcitriol in the regulation of macrophages and cancer cells in the TME and its influence on the proliferation of breast cancer cells. METHODS: We modeled the TME, in vitro, by collecting conditioned medium from cancer cells (CCM) and macrophages (MCM) and culturing each cell type separately with and without (control) a high-dose (0.5 µM) calcitriol (an active form of vitamin D). An MTT assay was used to examine cell viability. Apoptosis was detected using FITC (fluorescein isothiocyanate) annexin V apoptosis detection kit. Western blotting was used to separate and identify proteins. Quantitative real-time PCR was used to analyze gene expression. Molecular docking studies were performed to evaluate the binding type and interactions of calcitriol to the GLUT1 and mTORC1 ligand-binding sites. RESULTS: Calcitriol treatment suppressed the expression of genes and proteins implicated in glycolysis (GLUT1, HKII, LDHA), promoted cancer cell apoptosis, and reduced viability and Cyclin D1gene expression in MCM-induced breast cancer cells. Additionally, calcitriol treatment suppressed mTOR activation in MCM-induced breast cancer cells. Molecular docking studies further showed efficient binding of calcitriol with GLUT1 and mTORC1. Calcitriol also inhibited CCM-mediated induction of CD206 and increased TNFα gene expression in THP1-derived macrophages. CONCLUSION: The results suggest that calcitriol may impact breast cancer progression by inhibiting glycolysis and M2 macrophage polarization via regulating mTOR activation in the TME and warrants further investigation in vivo.
KW - Calcitriol; Tumor microenvironment; Macrophage polarization; Glycolysis; Apoptosis
KW - Breast Neoplasms/pathology
KW - Humans
KW - Macrophages/metabolism
KW - Mechanistic Target of Rapamycin Complex 1/metabolism
KW - Glucose Transporter Type 1/genetics
KW - Macrophage Activation
KW - Glycolysis
KW - Cell Line, Tumor
KW - Female
KW - Cell Proliferation/genetics
KW - Molecular Docking Simulation
KW - Tumor Microenvironment/genetics
KW - Calcitriol/pharmacology
KW - TOR Serine-Threonine Kinases/metabolism
UR - http://www.scopus.com/inward/record.url?scp=85152310845&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85152310845&partnerID=8YFLogxK
U2 - 10.33594/000000618
DO - 10.33594/000000618
M3 - Article
C2 - 37052042
AN - SCOPUS:85152310845
SN - 1015-8987
VL - 57
SP - 105
EP - 122
JO - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
JF - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
IS - 2
ER -