Pterostilbene-Isothiocyanate Inhibits Proliferation of Human MG-63 Osteosarcoma Cells Via Abrogating β-Catenin/TCF-4 Interaction-A Mechanistic Insight

Overview

Journal ACS Omega

Publisher American Chemical Society

Specialty Chemistry

Date 2023 Nov 29

PMID 38027335

Authors

Viney Kumar

Swati Haldar

Souvik Ghosh

Saakshi Saini

Poonam Dhankhar

Partha Roy

Affiliations

Soon will be listed here.

Abstract

Osteosarcoma, a highly metastasizing bone neoplasm, is a leading cause of death and disability in children and adolescents worldwide. Osteosarcoma is only suboptimally responsive to surgery and radio- and chemotherapy, that too with adverse side effects. Hence, there is a necessary need for safer alternative therapeutic approaches. This study evaluated the anticancer effects of the semi-synthetic compound, pterostilbene-isothiocyanate (PTER-ITC), on human osteosarcoma MG-63 cells through cytotoxicity, wound-healing, and transwell-migration assays. Results showed that PTER-ITC specifically inhibited the survival, proliferation, and migration of osteosarcoma cells. PTER-ITC induced apoptosis in MG-63 cells by disrupting mitochondrial membrane potential, as evident from the outcomes of different cytological staining. The antimetastatic potential of PTER-ITC was evaluated through immunostaining, RT-qPCR, and immunoblotting. (molecular docking and dynamic simulation) and, subsequently, biochemical [co-immunoprecipitation (Co-IP) and luciferase reporter] assays deciphered the underlying mode-of-action of this compound. PTER-ITC increased E-cadherin and reduced N-cadherin levels, thereby facilitating the reversal of epithelial-mesenchymal transition (EMT). It also modulated the expressions of proliferative cell nuclear antigen (PCNA), caspase-3, poly [ADP-ribose] polymerase (PARP-1) and matrix metalloproteinase-2/9 (MMPs-2/9) at transcriptional and translational levels. PTER-ITC interfered with the β-catenin/transcription factor-4 (TCF-4) interaction by occupying the β-catenin binding site on TCF-4, confirmed by their reduced physical interactions (Co-IP assay). This inhibited transcriptional activation of TCF-4 by β-catenin (as shown by luciferase reporter assay). In conclusion, PTER-ITC exhibited potent anticancer effects against human osteosarcoma cells by abrogating the β-catenin/TCF-4 interaction. Altogether, this study suggests that PTER-ITC may be regarded as a new approach for osteosarcoma treatment.

References

Park J, Sung I, Lee S, Kim K, Kim Y, Yoo M . The zinc-finger transcription factor Snail downregulates proliferating cell nuclear antigen expression in colorectal carcinoma cells. Int J Oncol. 2005; 26(6):1541-7. View

Shang S, Hua F, Hu Z . The regulation of β-catenin activity and function in cancer: therapeutic opportunities. Oncotarget. 2017; 8(20):33972-33989. PMC: 5464927. DOI: 10.18632/oncotarget.15687. View

Yang S, Chen J, Tan T, Wang N, Huang Y, Wang Y . Evodiamine Exerts Anticancer Effects Against 143B and MG63 Cells Through the Wnt/β-Catenin Signaling Pathway. Cancer Manag Res. 2020; 12:2875-2888. PMC: 7196244. DOI: 10.2147/CMAR.S238093. View

J Centelles J . General aspects of colorectal cancer. ISRN Oncol. 2012; 2012:139268. PMC: 3504424. DOI: 10.5402/2012/139268. View

Miyoshi A, Kitajima Y, Kido S, Shimonishi T, Matsuyama S, Kitahara K . Snail accelerates cancer invasion by upregulating MMP expression and is associated with poor prognosis of hepatocellular carcinoma. Br J Cancer. 2005; 92(2):252-8. PMC: 2361838. DOI: 10.1038/sj.bjc.6602266. View

Tran T, Ramasamy T, Choi J, Nguyen H, Pham T, Jeong J . Tumor-targeting, pH-sensitive nanoparticles for docetaxel delivery to drug-resistant cancer cells. Int J Nanomedicine. 2015; 10:5249-62. PMC: 4552257. DOI: 10.2147/IJN.S89584. View

Yan M, Li G, An J . Discovery of small molecule inhibitors of the Wnt/β-catenin signaling pathway by targeting β-catenin/Tcf4 interactions. Exp Biol Med (Maywood). 2017; 242(11):1185-1197. PMC: 5478005. DOI: 10.1177/1535370217708198. View

Yu W, Xu Z, Yuan L, Mo S, Xu B, Cheng X . Targeting β-Catenin Signaling by Natural Products for Cancer Prevention and Therapy. Front Pharmacol. 2020; 11:984. PMC: 7338604. DOI: 10.3389/fphar.2020.00984. View

Garrido C, Galluzzi L, Brunet M, Puig P, Didelot C, Kroemer G . Mechanisms of cytochrome c release from mitochondria. Cell Death Differ. 2006; 13(9):1423-33. DOI: 10.1038/sj.cdd.4401950. View

10.

Schneider C, Rasband W, Eliceiri K . NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012; 9(7):671-5. PMC: 5554542. DOI: 10.1038/nmeth.2089. View

11.

Zhang Y, Wang X . Targeting the Wnt/β-catenin signaling pathway in cancer. J Hematol Oncol. 2020; 13(1):165. PMC: 7716495. DOI: 10.1186/s13045-020-00990-3. View

12.

van der Spoel D, Lindahl E, Hess B, Groenhof G, Mark A, Berendsen H . GROMACS: fast, flexible, and free. J Comput Chem. 2005; 26(16):1701-18. DOI: 10.1002/jcc.20291. View

13.

Delgado-Deida Y, Alula K, Theiss A . The influence of mitochondrial-directed regulation of Wnt signaling on tumorigenesis. Gastroenterol Rep (Oxf). 2020; 8(3):215-223. PMC: 7333924. DOI: 10.1093/gastro/goaa025. View

14.

Isakoff M, Bielack S, Meltzer P, Gorlick R . Osteosarcoma: Current Treatment and a Collaborative Pathway to Success. J Clin Oncol. 2015; 33(27):3029-35. PMC: 4979196. DOI: 10.1200/JCO.2014.59.4895. View

15.

Newman D, Cragg G . Natural Products as Sources of New Drugs from 1981 to 2014. J Nat Prod. 2016; 79(3):629-61. DOI: 10.1021/acs.jnatprod.5b01055. View

16.

Nikhil K, Sharan S, Palla S, Sondhi S, Peddinti R, Roy P . Understanding the mode of action of a pterostilbene derivative as anti-inflammatory agent. Int Immunopharmacol. 2015; 28(1):10-21. DOI: 10.1016/j.intimp.2015.05.003. View

17.

Wu W, You R, Cheng C, Lee M, Lin T, Hu C . Snail collaborates with EGR-1 and SP-1 to directly activate transcription of MMP 9 and ZEB1. Sci Rep. 2017; 7(1):17753. PMC: 5736704. DOI: 10.1038/s41598-017-18101-7. View

18.

POY F, Lepourcelet M, Shivdasani R, Eck M . Structure of a human Tcf4-beta-catenin complex. Nat Struct Biol. 2001; 8(12):1053-7. DOI: 10.1038/nsb720. View

19.

Parisi A, Lacour F, Giordani L, Colnot S, Maire P, Le Grand F . APC is required for muscle stem cell proliferation and skeletal muscle tissue repair. J Cell Biol. 2015; 210(5):717-26. PMC: 4555822. DOI: 10.1083/jcb.201501053. View

20.

Park H, Park Y . Expression of p53 protein, PCNA, and Ki-67 in osteosarcomas of bone. J Korean Med Sci. 1995; 10(5):360-7. PMC: 3054151. DOI: 10.3346/jkms.1995.10.5.360. View